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El-Khodor)] TJ ET 0.271 0.267 0.267 rg BT 326.472 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 331.893 663.247 Td /F1 9.8 Tf [(Igor Filippov)] TJ ET 0.271 0.267 0.267 rg BT 384.991 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 390.412 663.247 Td /F1 9.8 Tf [(Richard Mushlin)] TJ ET 0.271 0.267 0.267 rg BT 460.300 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 465.721 663.247 Td /F1 9.8 Tf [(Russell Port)] TJ ET 0.271 0.267 0.267 rg BT 518.274 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 523.695 663.247 Td /F1 9.8 Tf [(David )] TJ ET BT 26.250 651.342 Td /F1 9.8 Tf [(Connor)] TJ ET 0.271 0.267 0.267 rg BT 58.220 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 63.641 651.342 Td /F1 9.8 Tf [(Ahmad Paintdakhi)] TJ ET 0.271 0.267 0.267 rg BT 142.762 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 148.184 651.342 Td /F1 9.8 Tf [(Liliana Menalled)] TJ ET 0.271 0.267 0.267 rg BT 218.627 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 224.048 651.342 Td /F1 9.8 Tf [(Sylvie Ramboz)] TJ ET 0.271 0.267 0.267 rg BT 289.061 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 294.482 651.342 Td /F1 9.8 Tf [(David Howland)] TJ ET 0.271 0.267 0.267 rg BT 360.041 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 365.462 651.342 Td /F1 9.8 Tf [(Seung Kwak)] TJ ET 0.271 0.267 0.267 rg BT 420.199 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 425.620 651.342 Td /F1 9.8 Tf [(Dani Brunner)] TJ ET 0.271 0.267 0.267 rg BT 26.250 639.438 Td /F1 9.8 Tf [(Balci F, Oakeshott S, Shamy JL, El-Khodor BF, Filippov I, Mushlin R, Port R, Connor D, Paintdakhi A, Menalled L, Ramboz S, )] TJ ET BT 26.250 627.533 Td /F1 9.8 Tf [(Howland D, Kwak S, Brunner D. High-Throughput Automated Phenotyping of Two Genetic Mouse Models of Huntingtons )] TJ ET BT 26.250 615.628 Td /F1 9.8 Tf [(Disease. PLOS Currents Huntington Disease. 2013 Jul 11 . Edition 1. doi: )] TJ ET BT 26.250 603.723 Td /F1 9.8 Tf [(10.1371/currents.hd.124aa0d16753f88215776fba102ceb29.)] TJ ET q 15.000 26.911 577.500 574.432 re W n 0.271 0.267 0.267 rg BT 26.250 574.621 Td /F4 12.0 Tf [(Abstract)] TJ ET BT 26.250 554.667 Td /F1 9.8 Tf [(Phenotyping with traditional behavioral assays constitutes a major bottleneck in the primary screening, characterization, and )] TJ ET BT 26.250 542.762 Td /F1 9.8 Tf [(validation of genetic mouse models of disease, leading to downstream delays in drug discovery efforts. We present a novel and )] TJ ET BT 26.250 530.857 Td /F1 9.8 Tf [(comprehensive one-stop approach to phenotyping, the PhenoCube. This system simultaneously captures the cognitive )] TJ ET BT 26.250 518.952 Td /F1 9.8 Tf [(performance, motor activity, and circadian patterns of group-housed mice by use of home-cage operant conditioning modules )] TJ ET BT 26.250 507.048 Td /F1 9.8 Tf [(\(IntelliCage\) and custom-built computer vision software. We evaluated two different mouse models of Huntingtons Disease )] TJ ET BT 26.250 495.143 Td /F1 9.8 Tf [(\(HD\), the R6/2 and the BACHD in the PhenoCube system. Our results demonstrated that this system can efficiently capture )] TJ ET BT 26.250 483.238 Td /F1 9.8 Tf [(and track alterations in both cognitive performance and locomotor activity patterns associated with these disease models. This )] TJ ET BT 26.250 471.333 Td /F1 9.8 Tf [(work extends our prior demonstration that PhenoCube can characterize circadian dysfunction in BACHD mice and shows that )] TJ ET BT 26.250 459.429 Td /F1 9.8 Tf [(this system, with the experimental protocols used, is a sensitive and efficient tool for a first pass high-throughput screening of )] TJ ET BT 26.250 447.524 Td /F1 9.8 Tf [(mouse disease models in general and mouse models of neurodegeneration in particular.)] TJ ET BT 26.250 410.921 Td /F4 12.0 Tf [(Funding Statement)] TJ ET BT 26.250 390.967 Td /F1 9.8 Tf [(Supported by the CHDI foundation. CHDI Foundation is a not-for-profit biomedical research organization exclusively dedicated )] TJ ET BT 26.250 379.062 Td /F1 9.8 Tf [(to discovering and developing therapeutics that slow the progression of Huntingtons disease. CHDI Foundation conducts )] TJ ET BT 26.250 367.158 Td /F1 9.8 Tf [(research in a number of different ways; for the purposes of this manuscript, all research was conceptualized, planned, and )] TJ ET BT 26.250 355.253 Td /F1 9.8 Tf [(directed by CHDI and PsychoGenics scientific staff and conducted at the contract research organization PsychoGenics, Inc. )] TJ ET BT 26.250 343.348 Td /F1 9.8 Tf [(Igor Filippov, Richard Mushlin, Ahmad Paintdakhi, Liliana Menalled, Sylvie Ramboz, and Dani Brunner are all employed by )] TJ ET BT 26.250 331.443 Td /F1 9.8 Tf [(PsychoGenics, Inc., a for-profit institution. The authors have declared that no further competing interests exist.)] TJ ET BT 26.250 302.341 Td /F4 12.0 Tf [(Introduction)] TJ ET BT 26.250 282.387 Td /F1 9.8 Tf [(Mouse models of human disease have provided valuable insights into our understanding of human genetic neurological )] TJ ET BT 26.250 270.482 Td /F1 9.8 Tf [(disorders. Generally, first pass validation of animal models is conducted using high throughput assays, in order to rapidly make )] TJ ET BT 26.250 258.577 Td /F1 9.8 Tf [(decisions about which models and compounds to pursue more in-depth. Other more time-consuming assays are a better fit for )] TJ ET BT 26.250 246.672 Td /F1 9.8 Tf [(secondary and tertiary tier testing, which is generally built upon and guided by results of first-order screening and )] TJ ET BT 26.250 234.768 Td /F1 9.8 Tf [(characterization.)] TJ ET BT 26.250 215.363 Td /F1 9.8 Tf [(Molecular, neuroimaging, neuropathological, and behavioral phenotyping are complementary approaches used together to )] TJ ET BT 26.250 203.458 Td /F1 9.8 Tf [(validate animal models of human disease and to evaluate potential therapeutics. Whereas there are many high-throughput )] TJ ET BT 26.250 191.553 Td /F1 9.8 Tf [(assays for the first three approaches, the options for high-throughput behavioral phenotyping are rather limited, with the result )] TJ ET BT 26.250 179.649 Td /F1 9.8 Tf [(that traditional behavioral tests are still standard practice for first-order validation of animal models. However such testing is )] TJ ET BT 26.250 167.744 Td /F1 9.8 Tf [(frequently slow, labor intensive, difficult to implement and can be vulnerable to subjective factors during experimentation and/or )] TJ ET BT 26.250 155.839 Td /F1 9.8 Tf [(scoring. This traditional approach typically involves a series of sequential tests of one aspect of behavior at a time limiting the )] TJ ET BT 26.250 143.934 Td /F1 9.8 Tf [(ability of these assays to capture the complexity and diversity of the behavioral phenotype of the animals across time. Such )] TJ ET BT 26.250 132.030 Td /F1 9.8 Tf [(tests often require extensive experimenter handling, making it hard to study genetically manipulated animals in non-stressed )] TJ ET BT 26.250 120.125 Td /F1 9.8 Tf [(conditions. It is possible that an ethologically valid environment with less experimenter intervention may be more reflective of the )] TJ ET BT 26.250 108.220 Td /F1 9.8 Tf [(natural state of the animal model and less susceptible to cross-laboratory discrepancies. Recognition of these limitations has led )] TJ ET BT 26.250 96.315 Td /F1 9.8 Tf [(several research laboratories and vendors to focus on automated approaches in behavioral phenotyping: e.g., PhenoMaster )] TJ ET BT 26.250 84.411 Td /F1 9.8 Tf [(\(TSE-Systems; )] TJ ET 0.267 0.267 0.267 rg BT 94.510 85.918 Td /F4 8.7 Tf [(1)] TJ ET 0.271 0.267 0.267 rg BT 99.328 84.411 Td /F1 9.8 Tf [(\), IntelliCage )] TJ ET 0.267 0.267 0.267 rg BT 156.766 85.918 Td /F4 8.7 Tf [(2)] TJ ET 0.271 0.267 0.267 rg BT 161.584 84.411 Td /F1 9.8 Tf [(, HomeCage Scan \(Cleversys, Inc.\) and fully-automated live-in environment )] TJ ET 0.267 0.267 0.267 rg BT 496.438 85.918 Td /F4 8.7 Tf [(3)] TJ ET 0.271 0.267 0.267 rg BT 501.257 84.411 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 65.006 Td /F1 9.8 Tf [(Here we provide a demonstration of our novel high-throughput phenotyping system, PhenoCube)] TJ ET BT 440.830 68.894 Td /F1 8.7 Tf [(TM)] TJ ET BT 453.344 65.006 Td /F1 9.8 Tf [(, based on the Intellicage )] TJ ET BT 26.250 53.101 Td /F1 9.8 Tf [(system, using two transgenic models of Huntingtons Disease \(HD\), the well-characterized R6/2 fragment mouse, and the full )] TJ ET BT 26.250 41.196 Td /F1 9.8 Tf [(length transgenic BACHD mouse developed in the laboratory of X. William Yang )] TJ ET 0.267 0.267 0.267 rg BT 374.686 42.704 Td /F4 8.7 Tf [(4)] TJ ET 0.271 0.267 0.267 rg BT 379.504 41.196 Td /F1 9.8 Tf [(. By high-throughput behavioral testing, we )] TJ ET Q q 15.000 684.354 577.500 53.646 re W n 0.267 0.267 0.267 rg BT 15.000 718.042 Td /F2 21.0 Tf [(High-Throughput Automated Phenotyping of Two Genetic )] TJ ET BT 15.000 693.094 Td /F2 21.0 Tf [(Mouse Models of Huntingtons Disease)] TJ ET Q 0.271 0.267 0.267 rg BT 15.000 675.088 Td /F3 9.8 Tf [(July 11, 2013)] TJ ET BT 72.681 675.088 Td /F3 9.8 Tf [()] TJ ET 0.267 0.267 0.267 rg BT 77.556 675.088 Td /F3 9.8 Tf [(HD Models)] TJ ET BT 26.250 663.247 Td /F1 9.8 Tf [(Fuat Balci)] TJ ET 0.271 0.267 0.267 rg BT 69.599 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 75.019 663.247 Td /F1 9.8 Tf [(Stephen Oakeshott)] TJ ET 0.271 0.267 0.267 rg BT 158.489 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 163.910 663.247 Td /F1 9.8 Tf [(Jul Lea Shamy)] TJ ET 0.271 0.267 0.267 rg BT 228.397 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 233.818 663.247 Td /F1 9.8 Tf [(Bassem F. El-Khodor)] TJ ET 0.271 0.267 0.267 rg BT 326.472 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 331.893 663.247 Td /F1 9.8 Tf [(Igor Filippov)] TJ ET 0.271 0.267 0.267 rg BT 384.991 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 390.412 663.247 Td /F1 9.8 Tf [(Richard Mushlin)] TJ ET 0.271 0.267 0.267 rg BT 460.300 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 465.721 663.247 Td /F1 9.8 Tf [(Russell Port)] TJ ET 0.271 0.267 0.267 rg BT 518.274 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 523.695 663.247 Td /F1 9.8 Tf [(David )] TJ ET BT 26.250 651.342 Td /F1 9.8 Tf [(Connor)] TJ ET 0.271 0.267 0.267 rg BT 58.220 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 63.641 651.342 Td /F1 9.8 Tf [(Ahmad Paintdakhi)] TJ ET 0.271 0.267 0.267 rg BT 142.762 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 148.184 651.342 Td /F1 9.8 Tf [(Liliana Menalled)] TJ ET 0.271 0.267 0.267 rg BT 218.627 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 224.048 651.342 Td /F1 9.8 Tf [(Sylvie Ramboz)] TJ ET 0.271 0.267 0.267 rg BT 289.061 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 294.482 651.342 Td /F1 9.8 Tf [(David Howland)] TJ ET 0.271 0.267 0.267 rg BT 360.041 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 365.462 651.342 Td /F1 9.8 Tf [(Seung Kwak)] TJ ET 0.271 0.267 0.267 rg BT 420.199 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 425.620 651.342 Td /F1 9.8 Tf [(Dani Brunner)] TJ ET 0.271 0.267 0.267 rg BT 26.250 639.438 Td /F1 9.8 Tf [(Balci F, Oakeshott S, Shamy JL, El-Khodor BF, Filippov I, Mushlin R, Port R, Connor D, Paintdakhi A, Menalled L, Ramboz S, )] TJ ET BT 26.250 627.533 Td /F1 9.8 Tf [(Howland D, Kwak S, Brunner D. High-Throughput Automated Phenotyping of Two Genetic Mouse Models of Huntingtons )] TJ ET BT 26.250 615.628 Td /F1 9.8 Tf [(Disease. PLOS Currents Huntington Disease. 2013 Jul 11 . Edition 1. doi: )] TJ ET BT 26.250 603.723 Td /F1 9.8 Tf [(10.1371/currents.hd.124aa0d16753f88215776fba102ceb29.)] TJ ET q 15.000 26.911 577.500 574.432 re W n 0.271 0.267 0.267 rg BT 26.250 574.621 Td /F4 12.0 Tf [(Abstract)] TJ ET BT 26.250 554.667 Td /F1 9.8 Tf [(Phenotyping with traditional behavioral assays constitutes a major bottleneck in the primary screening, characterization, and )] TJ ET BT 26.250 542.762 Td /F1 9.8 Tf [(validation of genetic mouse models of disease, leading to downstream delays in drug discovery efforts. We present a novel and )] TJ ET BT 26.250 530.857 Td /F1 9.8 Tf [(comprehensive one-stop approach to phenotyping, the PhenoCube. This system simultaneously captures the cognitive )] TJ ET BT 26.250 518.952 Td /F1 9.8 Tf [(performance, motor activity, and circadian patterns of group-housed mice by use of home-cage operant conditioning modules )] TJ ET BT 26.250 507.048 Td /F1 9.8 Tf [(\(IntelliCage\) and custom-built computer vision software. We evaluated two different mouse models of Huntingtons Disease )] TJ ET BT 26.250 495.143 Td /F1 9.8 Tf [(\(HD\), the R6/2 and the BACHD in the PhenoCube system. Our results demonstrated that this system can efficiently capture )] TJ ET BT 26.250 483.238 Td /F1 9.8 Tf [(and track alterations in both cognitive performance and locomotor activity patterns associated with these disease models. This )] TJ ET BT 26.250 471.333 Td /F1 9.8 Tf [(work extends our prior demonstration that PhenoCube can characterize circadian dysfunction in BACHD mice and shows that )] TJ ET BT 26.250 459.429 Td /F1 9.8 Tf [(this system, with the experimental protocols used, is a sensitive and efficient tool for a first pass high-throughput screening of )] TJ ET BT 26.250 447.524 Td /F1 9.8 Tf [(mouse disease models in general and mouse models of neurodegeneration in particular.)] TJ ET BT 26.250 410.921 Td /F4 12.0 Tf [(Funding Statement)] TJ ET BT 26.250 390.967 Td /F1 9.8 Tf [(Supported by the CHDI foundation. CHDI Foundation is a not-for-profit biomedical research organization exclusively dedicated )] TJ ET BT 26.250 379.062 Td /F1 9.8 Tf [(to discovering and developing therapeutics that slow the progression of Huntingtons disease. CHDI Foundation conducts )] TJ ET BT 26.250 367.158 Td /F1 9.8 Tf [(research in a number of different ways; for the purposes of this manuscript, all research was conceptualized, planned, and )] TJ ET BT 26.250 355.253 Td /F1 9.8 Tf [(directed by CHDI and PsychoGenics scientific staff and conducted at the contract research organization PsychoGenics, Inc. )] TJ ET BT 26.250 343.348 Td /F1 9.8 Tf [(Igor Filippov, Richard Mushlin, Ahmad Paintdakhi, Liliana Menalled, Sylvie Ramboz, and Dani Brunner are all employed by )] TJ ET BT 26.250 331.443 Td /F1 9.8 Tf [(PsychoGenics, Inc., a for-profit institution. The authors have declared that no further competing interests exist.)] TJ ET BT 26.250 302.341 Td /F4 12.0 Tf [(Introduction)] TJ ET BT 26.250 282.387 Td /F1 9.8 Tf [(Mouse models of human disease have provided valuable insights into our understanding of human genetic neurological )] TJ ET BT 26.250 270.482 Td /F1 9.8 Tf [(disorders. Generally, first pass validation of animal models is conducted using high throughput assays, in order to rapidly make )] TJ ET BT 26.250 258.577 Td /F1 9.8 Tf [(decisions about which models and compounds to pursue more in-depth. Other more time-consuming assays are a better fit for )] TJ ET BT 26.250 246.672 Td /F1 9.8 Tf [(secondary and tertiary tier testing, which is generally built upon and guided by results of first-order screening and )] TJ ET BT 26.250 234.768 Td /F1 9.8 Tf [(characterization.)] TJ ET BT 26.250 215.363 Td /F1 9.8 Tf [(Molecular, neuroimaging, neuropathological, and behavioral phenotyping are complementary approaches used together to )] TJ ET BT 26.250 203.458 Td /F1 9.8 Tf [(validate animal models of human disease and to evaluate potential therapeutics. Whereas there are many high-throughput )] TJ ET BT 26.250 191.553 Td /F1 9.8 Tf [(assays for the first three approaches, the options for high-throughput behavioral phenotyping are rather limited, with the result )] TJ ET BT 26.250 179.649 Td /F1 9.8 Tf [(that traditional behavioral tests are still standard practice for first-order validation of animal models. However such testing is )] TJ ET BT 26.250 167.744 Td /F1 9.8 Tf [(frequently slow, labor intensive, difficult to implement and can be vulnerable to subjective factors during experimentation and/or )] TJ ET BT 26.250 155.839 Td /F1 9.8 Tf [(scoring. This traditional approach typically involves a series of sequential tests of one aspect of behavior at a time limiting the )] TJ ET BT 26.250 143.934 Td /F1 9.8 Tf [(ability of these assays to capture the complexity and diversity of the behavioral phenotype of the animals across time. Such )] TJ ET BT 26.250 132.030 Td /F1 9.8 Tf [(tests often require extensive experimenter handling, making it hard to study genetically manipulated animals in non-stressed )] TJ ET BT 26.250 120.125 Td /F1 9.8 Tf [(conditions. It is possible that an ethologically valid environment with less experimenter intervention may be more reflective of the )] TJ ET BT 26.250 108.220 Td /F1 9.8 Tf [(natural state of the animal model and less susceptible to cross-laboratory discrepancies. Recognition of these limitations has led )] TJ ET BT 26.250 96.315 Td /F1 9.8 Tf [(several research laboratories and vendors to focus on automated approaches in behavioral phenotyping: e.g., PhenoMaster )] TJ ET BT 26.250 84.411 Td /F1 9.8 Tf [(\(TSE-Systems; )] TJ ET 0.267 0.267 0.267 rg BT 94.510 85.918 Td /F4 8.7 Tf [(1)] TJ ET 0.271 0.267 0.267 rg BT 99.328 84.411 Td /F1 9.8 Tf [(\), IntelliCage )] TJ ET 0.267 0.267 0.267 rg BT 156.766 85.918 Td /F4 8.7 Tf [(2)] TJ ET 0.271 0.267 0.267 rg BT 161.584 84.411 Td /F1 9.8 Tf [(, HomeCage Scan \(Cleversys, Inc.\) and fully-automated live-in environment )] TJ ET 0.267 0.267 0.267 rg BT 496.438 85.918 Td /F4 8.7 Tf [(3)] TJ ET 0.271 0.267 0.267 rg BT 501.257 84.411 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 65.006 Td /F1 9.8 Tf [(Here we provide a demonstration of our novel high-throughput phenotyping system, PhenoCube)] TJ ET BT 440.830 68.894 Td /F1 8.7 Tf [(TM)] TJ ET BT 453.344 65.006 Td /F1 9.8 Tf [(, based on the Intellicage )] TJ ET BT 26.250 53.101 Td /F1 9.8 Tf [(system, using two transgenic models of Huntingtons Disease \(HD\), the well-characterized R6/2 fragment mouse, and the full )] TJ ET BT 26.250 41.196 Td /F1 9.8 Tf [(length transgenic BACHD mouse developed in the laboratory of X. William Yang )] TJ ET 0.267 0.267 0.267 rg BT 374.686 42.704 Td /F4 8.7 Tf [(4)] TJ ET 0.271 0.267 0.267 rg BT 379.504 41.196 Td /F1 9.8 Tf [(. By high-throughput behavioral testing, we )] TJ ET Q q 15.000 684.354 577.500 53.646 re W n 0.267 0.267 0.267 rg BT 15.000 718.042 Td /F2 21.0 Tf [(High-Throughput Automated Phenotyping of Two Genetic )] TJ ET BT 15.000 693.094 Td /F2 21.0 Tf [(Mouse Models of Huntingtons Disease)] TJ ET Q 0.271 0.267 0.267 rg BT 15.000 675.088 Td /F3 9.8 Tf [(July 11, 2013)] TJ ET BT 72.681 675.088 Td /F3 9.8 Tf [()] TJ ET 0.267 0.267 0.267 rg BT 77.556 675.088 Td /F3 9.8 Tf [(HD Models)] TJ ET BT 26.250 663.247 Td /F1 9.8 Tf [(Fuat Balci)] TJ ET 0.271 0.267 0.267 rg BT 69.599 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 75.019 663.247 Td /F1 9.8 Tf [(Stephen Oakeshott)] TJ ET 0.271 0.267 0.267 rg BT 158.489 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 163.910 663.247 Td /F1 9.8 Tf [(Jul Lea Shamy)] TJ ET 0.271 0.267 0.267 rg BT 228.397 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 233.818 663.247 Td /F1 9.8 Tf [(Bassem F. El-Khodor)] TJ ET 0.271 0.267 0.267 rg BT 326.472 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 331.893 663.247 Td /F1 9.8 Tf [(Igor Filippov)] TJ ET 0.271 0.267 0.267 rg BT 384.991 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 390.412 663.247 Td /F1 9.8 Tf [(Richard Mushlin)] TJ ET 0.271 0.267 0.267 rg BT 460.300 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 465.721 663.247 Td /F1 9.8 Tf [(Russell Port)] TJ ET 0.271 0.267 0.267 rg BT 518.274 663.247 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 523.695 663.247 Td /F1 9.8 Tf [(David )] TJ ET BT 26.250 651.342 Td /F1 9.8 Tf [(Connor)] TJ ET 0.271 0.267 0.267 rg BT 58.220 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 63.641 651.342 Td /F1 9.8 Tf [(Ahmad Paintdakhi)] TJ ET 0.271 0.267 0.267 rg BT 142.762 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 148.184 651.342 Td /F1 9.8 Tf [(Liliana Menalled)] TJ ET 0.271 0.267 0.267 rg BT 218.627 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 224.048 651.342 Td /F1 9.8 Tf [(Sylvie Ramboz)] TJ ET 0.271 0.267 0.267 rg BT 289.061 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 294.482 651.342 Td /F1 9.8 Tf [(David Howland)] TJ ET 0.271 0.267 0.267 rg BT 360.041 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 365.462 651.342 Td /F1 9.8 Tf [(Seung Kwak)] TJ ET 0.271 0.267 0.267 rg BT 420.199 651.342 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 425.620 651.342 Td /F1 9.8 Tf [(Dani Brunner)] TJ ET 0.271 0.267 0.267 rg BT 26.250 639.438 Td /F1 9.8 Tf [(Balci F, Oakeshott S, Shamy JL, El-Khodor BF, Filippov I, Mushlin R, Port R, Connor D, Paintdakhi A, Menalled L, Ramboz S, )] TJ ET BT 26.250 627.533 Td /F1 9.8 Tf [(Howland D, Kwak S, Brunner D. High-Throughput Automated Phenotyping of Two Genetic Mouse Models of Huntingtons )] TJ ET BT 26.250 615.628 Td /F1 9.8 Tf [(Disease. PLOS Currents Huntington Disease. 2013 Jul 11 . Edition 1. doi: )] TJ ET BT 26.250 603.723 Td /F1 9.8 Tf [(10.1371/currents.hd.124aa0d16753f88215776fba102ceb29.)] TJ ET q 15.000 26.911 577.500 574.432 re W n 0.271 0.267 0.267 rg BT 26.250 574.621 Td /F4 12.0 Tf [(Abstract)] TJ ET BT 26.250 554.667 Td /F1 9.8 Tf [(Phenotyping with traditional behavioral assays constitutes a major bottleneck in the primary screening, characterization, and )] TJ ET BT 26.250 542.762 Td /F1 9.8 Tf [(validation of genetic mouse models of disease, leading to downstream delays in drug discovery efforts. We present a novel and )] TJ ET BT 26.250 530.857 Td /F1 9.8 Tf [(comprehensive one-stop approach to phenotyping, the PhenoCube. This system simultaneously captures the cognitive )] TJ ET BT 26.250 518.952 Td /F1 9.8 Tf [(performance, motor activity, and circadian patterns of group-housed mice by use of home-cage operant conditioning modules )] TJ ET BT 26.250 507.048 Td /F1 9.8 Tf [(\(IntelliCage\) and custom-built computer vision software. We evaluated two different mouse models of Huntingtons Disease )] TJ ET BT 26.250 495.143 Td /F1 9.8 Tf [(\(HD\), the R6/2 and the BACHD in the PhenoCube system. Our results demonstrated that this system can efficiently capture )] TJ ET BT 26.250 483.238 Td /F1 9.8 Tf [(and track alterations in both cognitive performance and locomotor activity patterns associated with these disease models. This )] TJ ET BT 26.250 471.333 Td /F1 9.8 Tf [(work extends our prior demonstration that PhenoCube can characterize circadian dysfunction in BACHD mice and shows that )] TJ ET BT 26.250 459.429 Td /F1 9.8 Tf [(this system, with the experimental protocols used, is a sensitive and efficient tool for a first pass high-throughput screening of )] TJ ET BT 26.250 447.524 Td /F1 9.8 Tf [(mouse disease models in general and mouse models of neurodegeneration in particular.)] TJ ET BT 26.250 410.921 Td /F4 12.0 Tf [(Funding Statement)] TJ ET BT 26.250 390.967 Td /F1 9.8 Tf [(Supported by the CHDI foundation. CHDI Foundation is a not-for-profit biomedical research organization exclusively dedicated )] TJ ET BT 26.250 379.062 Td /F1 9.8 Tf [(to discovering and developing therapeutics that slow the progression of Huntingtons disease. CHDI Foundation conducts )] TJ ET BT 26.250 367.158 Td /F1 9.8 Tf [(research in a number of different ways; for the purposes of this manuscript, all research was conceptualized, planned, and )] TJ ET BT 26.250 355.253 Td /F1 9.8 Tf [(directed by CHDI and PsychoGenics scientific staff and conducted at the contract research organization PsychoGenics, Inc. )] TJ ET BT 26.250 343.348 Td /F1 9.8 Tf [(Igor Filippov, Richard Mushlin, Ahmad Paintdakhi, Liliana Menalled, Sylvie Ramboz, and Dani Brunner are all employed by )] TJ ET BT 26.250 331.443 Td /F1 9.8 Tf [(PsychoGenics, Inc., a for-profit institution. The authors have declared that no further competing interests exist.)] TJ ET BT 26.250 302.341 Td /F4 12.0 Tf [(Introduction)] TJ ET BT 26.250 282.387 Td /F1 9.8 Tf [(Mouse models of human disease have provided valuable insights into our understanding of human genetic neurological )] TJ ET BT 26.250 270.482 Td /F1 9.8 Tf [(disorders. Generally, first pass validation of animal models is conducted using high throughput assays, in order to rapidly make )] TJ ET BT 26.250 258.577 Td /F1 9.8 Tf [(decisions about which models and compounds to pursue more in-depth. Other more time-consuming assays are a better fit for )] TJ ET BT 26.250 246.672 Td /F1 9.8 Tf [(secondary and tertiary tier testing, which is generally built upon and guided by results of first-order screening and )] TJ ET BT 26.250 234.768 Td /F1 9.8 Tf [(characterization.)] TJ ET BT 26.250 215.363 Td /F1 9.8 Tf [(Molecular, neuroimaging, neuropathological, and behavioral phenotyping are complementary approaches used together to )] TJ ET BT 26.250 203.458 Td /F1 9.8 Tf [(validate animal models of human disease and to evaluate potential therapeutics. Whereas there are many high-throughput )] TJ ET BT 26.250 191.553 Td /F1 9.8 Tf [(assays for the first three approaches, the options for high-throughput behavioral phenotyping are rather limited, with the result )] TJ ET BT 26.250 179.649 Td /F1 9.8 Tf [(that traditional behavioral tests are still standard practice for first-order validation of animal models. However such testing is )] TJ ET BT 26.250 167.744 Td /F1 9.8 Tf [(frequently slow, labor intensive, difficult to implement and can be vulnerable to subjective factors during experimentation and/or )] TJ ET BT 26.250 155.839 Td /F1 9.8 Tf [(scoring. This traditional approach typically involves a series of sequential tests of one aspect of behavior at a time limiting the )] TJ ET BT 26.250 143.934 Td /F1 9.8 Tf [(ability of these assays to capture the complexity and diversity of the behavioral phenotype of the animals across time. Such )] TJ ET BT 26.250 132.030 Td /F1 9.8 Tf [(tests often require extensive experimenter handling, making it hard to study genetically manipulated animals in non-stressed )] TJ ET BT 26.250 120.125 Td /F1 9.8 Tf [(conditions. It is possible that an ethologically valid environment with less experimenter intervention may be more reflective of the )] TJ ET BT 26.250 108.220 Td /F1 9.8 Tf [(natural state of the animal model and less susceptible to cross-laboratory discrepancies. Recognition of these limitations has led )] TJ ET BT 26.250 96.315 Td /F1 9.8 Tf [(several research laboratories and vendors to focus on automated approaches in behavioral phenotyping: e.g., PhenoMaster )] TJ ET BT 26.250 84.411 Td /F1 9.8 Tf [(\(TSE-Systems; )] TJ ET 0.267 0.267 0.267 rg BT 94.510 85.918 Td /F4 8.7 Tf [(1)] TJ ET 0.271 0.267 0.267 rg BT 99.328 84.411 Td /F1 9.8 Tf [(\), IntelliCage )] TJ ET 0.267 0.267 0.267 rg BT 156.766 85.918 Td /F4 8.7 Tf [(2)] TJ ET 0.271 0.267 0.267 rg BT 161.584 84.411 Td /F1 9.8 Tf [(, HomeCage Scan \(Cleversys, Inc.\) and fully-automated live-in environment )] TJ ET 0.267 0.267 0.267 rg BT 496.438 85.918 Td /F4 8.7 Tf [(3)] TJ ET 0.271 0.267 0.267 rg BT 501.257 84.411 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 65.006 Td /F1 9.8 Tf [(Here we provide a demonstration of our novel high-throughput phenotyping system, PhenoCube)] TJ ET BT 440.830 68.894 Td /F1 8.7 Tf [(TM)] TJ ET BT 453.344 65.006 Td /F1 9.8 Tf [(, based on the Intellicage )] TJ ET BT 26.250 53.101 Td /F1 9.8 Tf [(system, using two transgenic models of Huntingtons Disease \(HD\), the well-characterized R6/2 fragment mouse, and the full )] TJ ET BT 26.250 41.196 Td /F1 9.8 Tf [(length transgenic BACHD mouse developed in the laboratory of X. William Yang )] TJ ET 0.267 0.267 0.267 rg BT 374.686 42.704 Td /F4 8.7 Tf [(4)] TJ ET 0.271 0.267 0.267 rg BT 379.504 41.196 Td /F1 9.8 Tf [(. 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>> endobj 146 0 obj << /Type /Page /Parent 3 0 R /Annots [ 148 0 R 150 0 R 152 0 R 154 0 R 156 0 R 158 0 R 160 0 R 162 0 R 164 0 R 166 0 R 168 0 R 170 0 R 172 0 R 174 0 R 176 0 R 178 0 R 180 0 R 182 0 R 184 0 R 186 0 R 188 0 R 190 0 R 192 0 R 194 0 R 196 0 R 198 0 R 200 0 R 202 0 R 204 0 R 206 0 R 208 0 R 210 0 R 212 0 R 214 0 R 216 0 R 218 0 R 220 0 R 222 0 R 224 0 R 226 0 R 228 0 R 230 0 R 232 0 R 234 0 R 236 0 R 238 0 R 240 0 R 242 0 R 244 0 R 246 0 R 248 0 R 250 0 R 252 0 R 254 0 R 256 0 R 258 0 R 260 0 R 262 0 R 264 0 R 266 0 R 268 0 R 270 0 R 272 0 R 274 0 R 276 0 R 278 0 R 280 0 R 282 0 R 284 0 R 286 0 R 288 0 R 290 0 R 292 0 R 294 0 R 296 0 R ] /Contents 147 0 R >> endobj 147 0 obj << /Length 36262 >> stream 0.271 0.267 0.267 rg q 15.000 45.801 577.500 731.199 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(mean efficient and quick screening of animal subjects for multiple phenotypes simultaneously. These phenotypes might include )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(\(but are not limited to\) cognitive performance, motivation and locomotor, circadian, and exploratory activity. In addition, we )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(showed that the system has high sensitivity and robustness to detect the deficits in these mutant lines.)] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(The R6/2 mouse model expresses a short N-terminal fragment of mutant human huntingtin and is the best-known mouse model )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(of HD )] TJ ET 0.267 0.267 0.267 rg BT 53.882 713.864 Td /F4 8.7 Tf [(5)] TJ ET 0.271 0.267 0.267 rg BT 58.700 712.357 Td /F1 9.8 Tf [( in terms of pathophysiology and behavioral abnormalities \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 335.074 713.864 Td /F4 8.7 Tf [(7)] TJ ET 0.271 0.267 0.267 rg BT 339.892 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 342.302 713.864 Td /F4 8.7 Tf [(8)] TJ ET 0.271 0.267 0.267 rg BT 347.120 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 349.530 713.864 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 354.348 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 356.758 713.864 Td /F4 8.7 Tf [(10)] TJ ET 0.271 0.267 0.267 rg BT 366.395 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 368.804 713.864 Td /F4 8.7 Tf [(11)] TJ ET 0.271 0.267 0.267 rg BT 378.442 712.357 Td /F1 9.8 Tf [(\). R6/2 mice have been shown to be )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(hypoactive \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 100.496 701.960 Td /F4 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 110.134 704.341 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 112.543 701.960 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 117.362 700.452 Td /F1 9.8 Tf [(\), exhibit disrupted circadian patterns \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 304.318 701.960 Td /F4 8.7 Tf [(13)] TJ ET 0.271 0.267 0.267 rg BT 313.955 700.452 Td /F1 9.8 Tf [(\), response acquisition )] TJ ET 0.267 0.267 0.267 rg BT 414.205 701.960 Td /F4 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 423.842 700.452 Td /F1 9.8 Tf [( and interval timing )] TJ ET 0.267 0.267 0.267 rg BT 508.375 701.960 Td /F4 8.7 Tf [(15)] TJ ET 0.271 0.267 0.267 rg BT 518.012 700.452 Td /F1 9.8 Tf [(, and have )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(impaired spatial discrimination and reversal as assessed in water-maze and t-maze tasks \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 438.597 690.055 Td /F4 8.7 Tf [(8)] TJ ET 0.271 0.267 0.267 rg BT 443.416 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 445.825 690.055 Td /F4 8.7 Tf [(16)] TJ ET 0.271 0.267 0.267 rg BT 455.462 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 457.872 690.055 Td /F4 8.7 Tf [(17)] TJ ET 0.271 0.267 0.267 rg BT 467.509 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 469.918 690.055 Td /F4 8.7 Tf [(18)] TJ ET 0.271 0.267 0.267 rg BT 479.556 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 481.965 690.055 Td /F4 8.7 Tf [(19)] TJ ET 0.271 0.267 0.267 rg BT 491.602 688.548 Td /F1 9.8 Tf [(\). By contrast, the )] TJ ET BT 26.250 676.643 Td /F1 9.8 Tf [(BACHD mouse expresses the full-length human mutant huntingtin gene with 97 glutamine repeats under the control of )] TJ ET BT 26.250 664.738 Td /F1 9.8 Tf [(endogenous htt regulatory machinery on a bacterial artificial chromosome, and has been less widely studied but also has been )] TJ ET BT 26.250 652.833 Td /F1 9.8 Tf [(shown to mimic some aspects of HD pathophysiology and behavioral abnormalities )] TJ ET 0.267 0.267 0.267 rg BT 387.137 654.341 Td /F4 8.7 Tf [(4)] TJ ET 0.271 0.267 0.267 rg BT 391.955 656.722 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 394.365 654.341 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 399.183 656.722 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 401.593 654.341 Td /F4 8.7 Tf [(20)] TJ ET 0.271 0.267 0.267 rg BT 411.230 652.833 Td /F1 9.8 Tf [(, including circadian abnormalities )] TJ ET 0.267 0.267 0.267 rg BT 560.239 654.341 Td /F4 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 569.876 656.722 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 26.250 642.436 Td /F4 8.7 Tf [(21)] TJ ET 0.271 0.267 0.267 rg BT 35.887 640.929 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(The PhenoCube)] TJ ET BT 97.249 625.412 Td /F1 8.7 Tf [(TM)] TJ ET BT 109.764 621.524 Td /F1 9.8 Tf [( system acquires a broad range of different measures that span multiple disease-relevant domains \(i.e., )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(cognition, locomotor activity and circadian patterns\), and thus can efficiently capture the complexity of the behavioral phenotype. )] TJ ET BT 26.250 597.714 Td /F1 9.8 Tf [(We developed the PhenoCube)] TJ ET BT 159.035 601.603 Td /F1 8.7 Tf [(TM)] TJ ET BT 171.550 597.714 Td /F1 9.8 Tf [( through hardware modifications of IntelliCage units and addition of custom-built computer )] TJ ET BT 26.250 585.810 Td /F1 9.8 Tf [(vision hardware and software. This system enables behavioral phenotyping of group-housed mice within a home-cage-like )] TJ ET BT 26.250 573.905 Td /F1 9.8 Tf [(environment over multiple days with minimal experimenter interruption, allowing capture of the endogenous behavioral rhythms )] TJ ET BT 26.250 562.000 Td /F1 9.8 Tf [(of the subjects. Models of HD are particularly relevant disease model for demonstration of the utility of this system as it is )] TJ ET BT 26.250 550.095 Td /F1 9.8 Tf [(characterized by altered cognition, motor activity, and circadian rhythms, all domains that can all be assessed simultaneously by )] TJ ET BT 26.250 538.191 Td /F1 9.8 Tf [(the PhenoCube)] TJ ET BT 94.003 542.079 Td /F1 8.7 Tf [(TM)] TJ ET BT 106.517 538.191 Td /F1 9.8 Tf [(. The present experiments capture these behavioral phenotypes using the novel PhenoCube)] TJ ET BT 504.854 542.079 Td /F1 8.7 Tf [(TM)] TJ ET BT 517.368 538.191 Td /F1 9.8 Tf [( apparatus, )] TJ ET BT 26.250 526.286 Td /F1 9.8 Tf [(demonstrating the utility of the system and of the experimental protocols developed in our lab.)] TJ ET BT 26.250 489.683 Td /F4 12.0 Tf [(METHODS)] TJ ET BT 26.250 469.729 Td /F4 9.8 Tf [(Animals:)] TJ ET BT 26.250 450.324 Td /F1 9.8 Tf [(R6/2 \(R6/2 CAG 120, CHDI-81001000\) transgenic heterozygous and wild type \(WT\) littermate control mice were obtained from )] TJ ET BT 26.250 438.420 Td /F1 9.8 Tf [(Jackson Laboratories \(Bar Harbor,ME; strain B6CBA-Tg[HDexon1]62Gpb/3J, stock number 006494\). Mice were generated by )] TJ ET BT 26.250 426.515 Td /F1 9.8 Tf [(crossing ovarian-transplanted females \(from R6/2 CBAB6J female donors\) with CBAB6F1J WT males. Genotype was )] TJ ET BT 26.250 414.610 Td /F1 9.8 Tf [(determined by polymerase chain reaction \(PCR\) of tail-tip DNA at 15 days of age )] TJ ET 0.267 0.267 0.267 rg BT 377.387 416.117 Td /F4 8.7 Tf [(22)] TJ ET 0.271 0.267 0.267 rg BT 387.024 414.610 Td /F1 9.8 Tf [(. CAG repeat lengths were measured by )] TJ ET BT 26.250 402.705 Td /F1 9.8 Tf [(Laragen \(Los Angeles, CA, USA\) using standard protocols and Genemapper software as previously described )] TJ ET 0.267 0.267 0.267 rg BT 503.668 404.213 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 508.487 402.705 Td /F1 9.8 Tf [(. R6/2 mice )] TJ ET BT 26.250 390.801 Td /F1 9.8 Tf [(were tested at 7, 8, 9 and 10 weeks of age. The CAG repeat of the R6/2 mutant is 120 )] TJ ET BT 400.757 390.801 Td /F1 9.8 Tf [(+)] TJ ET BT 406.451 390.801 Td /F1 9.8 Tf [( 5.)] TJ ET BT 26.250 371.396 Td /F1 9.8 Tf [(BACHD \(CHDI-81001012and WT control mice were generated at the Jackson laboratory on an FVB/NJ x C57Bl/6J F1, created )] TJ ET BT 26.250 359.491 Td /F1 9.8 Tf [(by crossing BAC transgenic FVB/NJ males with WT C57Bl/6J females. The congenic BAC FVB/NJ line used for breeding was )] TJ ET BT 26.250 347.586 Td /F1 9.8 Tf [(obtained from the laboratory of X. William Yang at the UCLA David Geffen School of Medicine, Los Angeles. Genotyping was )] TJ ET BT 26.250 335.682 Td /F1 9.8 Tf [(completed by Laragen \(Los Angeles, CA, USA\). BACHD and the corresponding WT control mice were tested at 16, 24, 36, and )] TJ ET BT 26.250 323.777 Td /F1 9.8 Tf [(52 weeks of age.)] TJ ET BT 26.250 304.372 Td /F1 9.8 Tf [(Prior to testing in PhenoCube)] TJ ET BT 153.605 308.260 Td /F1 8.7 Tf [(TM)] TJ ET BT 166.119 304.372 Td /F1 9.8 Tf [(, mice were injected with sterile transponders \(T-IS 8010 FDX-B, Datamars SA, Bedano, )] TJ ET BT 26.250 292.467 Td /F1 9.8 Tf [(Switzerland\) under 2% isoflurane inhalation anesthesia. During recovery from anesthesia, mice were introduced to their social )] TJ ET BT 26.250 280.563 Td /F1 9.8 Tf [(testing groups and housed in genetically homogenous in OptiRAT cages \(Animal Care Systems, CO\). The environment was )] TJ ET BT 26.250 268.658 Td /F1 9.8 Tf [(enriched with a play tunnel, shredded paper, and a plastic bone. Food and water were available ad libitum with the exception of )] TJ ET BT 26.250 256.753 Td /F1 9.8 Tf [(water deprivation 16 hours prior to each phase of testing in our system. Temperature and humidity were controlled and )] TJ ET BT 26.250 244.848 Td /F1 9.8 Tf [(monitored daily. This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of )] TJ ET BT 26.250 232.944 Td /F1 9.8 Tf [(Laboratory Animals, NRC )] TJ ET 0.267 0.267 0.267 rg BT 139.487 234.451 Td /F4 8.7 Tf [(23)] TJ ET 0.271 0.267 0.267 rg BT 149.124 232.944 Td /F1 9.8 Tf [(. The protocol was approved by the Institutional Animal Care and Use Committee of Psychogenics, )] TJ ET BT 26.250 221.039 Td /F1 9.8 Tf [(Inc. \(PHS OLAW animal welfare assurance number A4471-01\), an AAALAC International accredited institution \(Unit #001213\).)] TJ ET BT 26.250 201.634 Td /F4 9.8 Tf [(Apparatus:)] TJ ET BT 26.250 182.229 Td /F1 9.8 Tf [(Experiments were conducted using modified Intellicage units \(New Behavior, AG, Zurich, CH\), each with a Day/Night Camera )] TJ ET BT 26.250 170.325 Td /F1 9.8 Tf [(mounted on top of the cage for Computer Vision \(CV\) analysis \(Figure 1\). Intra-maze spatial cues were added to the )] TJ ET BT 26.250 158.420 Td /F1 9.8 Tf [(environment by placing laminated striped paper on the outside of the cage walls, while three climbing structures \(two rods, a )] TJ ET BT 26.250 146.515 Td /F1 9.8 Tf [(cubic central object and a three step staircase\) were placed inside the cage to provide an enriched topology \(Figure 1 left )] TJ ET BT 26.250 134.610 Td /F1 9.8 Tf [(panel\).)] TJ ET BT 26.250 115.206 Td /F1 9.8 Tf [(The cages were maintained at all times on a 12:12 light/dark cycle, with white light during the day and red light during the night. )] TJ ET BT 26.250 103.301 Td /F1 9.8 Tf [(The light intensity under red light was recorded at 7 lux using a photographic band-pass filter \(LDP LLC, NJ\) that eliminates long )] TJ ET BT 26.250 91.396 Td /F1 9.8 Tf [(wavelength light frequencies not visible to mice )] TJ ET 0.267 0.267 0.267 rg BT 232.170 92.903 Td /F4 8.7 Tf [(24)] TJ ET 0.271 0.267 0.267 rg BT 241.807 91.396 Td /F1 9.8 Tf [( and was sufficient to allow the camera to detect the mice while maintaining a )] TJ ET BT 26.250 79.491 Td /F1 9.8 Tf [(low subjective light level for the mice. The light intensity under white light was 100 lux. Inside the PhenoCube)] TJ ET BT 495.001 83.380 Td /F1 8.7 Tf [(TM)] TJ ET BT 507.515 79.491 Td /F1 9.8 Tf [( environment, )] TJ ET BT 26.250 67.587 Td /F1 9.8 Tf [(water was only available from within the corners while food was freely available on the cage floor at all times. It was sometimes )] TJ ET BT 26.250 55.682 Td /F1 9.8 Tf [(necessary to remove a mouse from the study when no licking was recorded, in order to ensure proper hydration.)] TJ ET Q q 15.000 45.801 577.500 731.199 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(mean efficient and quick screening of animal subjects for multiple phenotypes simultaneously. These phenotypes might include )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(\(but are not limited to\) cognitive performance, motivation and locomotor, circadian, and exploratory activity. In addition, we )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(showed that the system has high sensitivity and robustness to detect the deficits in these mutant lines.)] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(The R6/2 mouse model expresses a short N-terminal fragment of mutant human huntingtin and is the best-known mouse model )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(of HD )] TJ ET 0.267 0.267 0.267 rg BT 53.882 713.864 Td /F4 8.7 Tf [(5)] TJ ET 0.271 0.267 0.267 rg BT 58.700 712.357 Td /F1 9.8 Tf [( in terms of pathophysiology and behavioral abnormalities \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 335.074 713.864 Td /F4 8.7 Tf [(7)] TJ ET 0.271 0.267 0.267 rg BT 339.892 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 342.302 713.864 Td /F4 8.7 Tf [(8)] TJ ET 0.271 0.267 0.267 rg BT 347.120 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 349.530 713.864 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 354.348 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 356.758 713.864 Td /F4 8.7 Tf [(10)] TJ ET 0.271 0.267 0.267 rg BT 366.395 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 368.804 713.864 Td /F4 8.7 Tf [(11)] TJ ET 0.271 0.267 0.267 rg BT 378.442 712.357 Td /F1 9.8 Tf [(\). R6/2 mice have been shown to be )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(hypoactive \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 100.496 701.960 Td /F4 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 110.134 704.341 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 112.543 701.960 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 117.362 700.452 Td /F1 9.8 Tf [(\), exhibit disrupted circadian patterns \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 304.318 701.960 Td /F4 8.7 Tf [(13)] TJ ET 0.271 0.267 0.267 rg BT 313.955 700.452 Td /F1 9.8 Tf [(\), response acquisition )] TJ ET 0.267 0.267 0.267 rg BT 414.205 701.960 Td /F4 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 423.842 700.452 Td /F1 9.8 Tf [( and interval timing )] TJ ET 0.267 0.267 0.267 rg BT 508.375 701.960 Td /F4 8.7 Tf [(15)] TJ ET 0.271 0.267 0.267 rg BT 518.012 700.452 Td /F1 9.8 Tf [(, and have )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(impaired spatial discrimination and reversal as assessed in water-maze and t-maze tasks \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 438.597 690.055 Td /F4 8.7 Tf [(8)] TJ ET 0.271 0.267 0.267 rg BT 443.416 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 445.825 690.055 Td /F4 8.7 Tf [(16)] TJ ET 0.271 0.267 0.267 rg BT 455.462 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 457.872 690.055 Td /F4 8.7 Tf [(17)] TJ ET 0.271 0.267 0.267 rg BT 467.509 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 469.918 690.055 Td /F4 8.7 Tf [(18)] TJ ET 0.271 0.267 0.267 rg BT 479.556 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 481.965 690.055 Td /F4 8.7 Tf [(19)] TJ ET 0.271 0.267 0.267 rg BT 491.602 688.548 Td /F1 9.8 Tf [(\). By contrast, the )] TJ ET BT 26.250 676.643 Td /F1 9.8 Tf [(BACHD mouse expresses the full-length human mutant huntingtin gene with 97 glutamine repeats under the control of )] TJ ET BT 26.250 664.738 Td /F1 9.8 Tf [(endogenous htt regulatory machinery on a bacterial artificial chromosome, and has been less widely studied but also has been )] TJ ET BT 26.250 652.833 Td /F1 9.8 Tf [(shown to mimic some aspects of HD pathophysiology and behavioral abnormalities )] TJ ET 0.267 0.267 0.267 rg BT 387.137 654.341 Td /F4 8.7 Tf [(4)] TJ ET 0.271 0.267 0.267 rg BT 391.955 656.722 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 394.365 654.341 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 399.183 656.722 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 401.593 654.341 Td /F4 8.7 Tf [(20)] TJ ET 0.271 0.267 0.267 rg BT 411.230 652.833 Td /F1 9.8 Tf [(, including circadian abnormalities )] TJ ET 0.267 0.267 0.267 rg BT 560.239 654.341 Td /F4 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 569.876 656.722 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 26.250 642.436 Td /F4 8.7 Tf [(21)] TJ ET 0.271 0.267 0.267 rg BT 35.887 640.929 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(The PhenoCube)] TJ ET BT 97.249 625.412 Td /F1 8.7 Tf [(TM)] TJ ET BT 109.764 621.524 Td /F1 9.8 Tf [( system acquires a broad range of different measures that span multiple disease-relevant domains \(i.e., )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(cognition, locomotor activity and circadian patterns\), and thus can efficiently capture the complexity of the behavioral phenotype. )] TJ ET BT 26.250 597.714 Td /F1 9.8 Tf [(We developed the PhenoCube)] TJ ET BT 159.035 601.603 Td /F1 8.7 Tf [(TM)] TJ ET BT 171.550 597.714 Td /F1 9.8 Tf [( through hardware modifications of IntelliCage units and addition of custom-built computer )] TJ ET BT 26.250 585.810 Td /F1 9.8 Tf [(vision hardware and software. This system enables behavioral phenotyping of group-housed mice within a home-cage-like )] TJ ET BT 26.250 573.905 Td /F1 9.8 Tf [(environment over multiple days with minimal experimenter interruption, allowing capture of the endogenous behavioral rhythms )] TJ ET BT 26.250 562.000 Td /F1 9.8 Tf [(of the subjects. Models of HD are particularly relevant disease model for demonstration of the utility of this system as it is )] TJ ET BT 26.250 550.095 Td /F1 9.8 Tf [(characterized by altered cognition, motor activity, and circadian rhythms, all domains that can all be assessed simultaneously by )] TJ ET BT 26.250 538.191 Td /F1 9.8 Tf [(the PhenoCube)] TJ ET BT 94.003 542.079 Td /F1 8.7 Tf [(TM)] TJ ET BT 106.517 538.191 Td /F1 9.8 Tf [(. The present experiments capture these behavioral phenotypes using the novel PhenoCube)] TJ ET BT 504.854 542.079 Td /F1 8.7 Tf [(TM)] TJ ET BT 517.368 538.191 Td /F1 9.8 Tf [( apparatus, )] TJ ET BT 26.250 526.286 Td /F1 9.8 Tf [(demonstrating the utility of the system and of the experimental protocols developed in our lab.)] TJ ET BT 26.250 489.683 Td /F4 12.0 Tf [(METHODS)] TJ ET BT 26.250 469.729 Td /F4 9.8 Tf [(Animals:)] TJ ET BT 26.250 450.324 Td /F1 9.8 Tf [(R6/2 \(R6/2 CAG 120, CHDI-81001000\) transgenic heterozygous and wild type \(WT\) littermate control mice were obtained from )] TJ ET BT 26.250 438.420 Td /F1 9.8 Tf [(Jackson Laboratories \(Bar Harbor,ME; strain B6CBA-Tg[HDexon1]62Gpb/3J, stock number 006494\). Mice were generated by )] TJ ET BT 26.250 426.515 Td /F1 9.8 Tf [(crossing ovarian-transplanted females \(from R6/2 CBAB6J female donors\) with CBAB6F1J WT males. Genotype was )] TJ ET BT 26.250 414.610 Td /F1 9.8 Tf [(determined by polymerase chain reaction \(PCR\) of tail-tip DNA at 15 days of age )] TJ ET 0.267 0.267 0.267 rg BT 377.387 416.117 Td /F4 8.7 Tf [(22)] TJ ET 0.271 0.267 0.267 rg BT 387.024 414.610 Td /F1 9.8 Tf [(. CAG repeat lengths were measured by )] TJ ET BT 26.250 402.705 Td /F1 9.8 Tf [(Laragen \(Los Angeles, CA, USA\) using standard protocols and Genemapper software as previously described )] TJ ET 0.267 0.267 0.267 rg BT 503.668 404.213 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 508.487 402.705 Td /F1 9.8 Tf [(. R6/2 mice )] TJ ET BT 26.250 390.801 Td /F1 9.8 Tf [(were tested at 7, 8, 9 and 10 weeks of age. The CAG repeat of the R6/2 mutant is 120 )] TJ ET BT 400.757 390.801 Td /F1 9.8 Tf [(+)] TJ ET BT 406.451 390.801 Td /F1 9.8 Tf [( 5.)] TJ ET BT 26.250 371.396 Td /F1 9.8 Tf [(BACHD \(CHDI-81001012and WT control mice were generated at the Jackson laboratory on an FVB/NJ x C57Bl/6J F1, created )] TJ ET BT 26.250 359.491 Td /F1 9.8 Tf [(by crossing BAC transgenic FVB/NJ males with WT C57Bl/6J females. The congenic BAC FVB/NJ line used for breeding was )] TJ ET BT 26.250 347.586 Td /F1 9.8 Tf [(obtained from the laboratory of X. William Yang at the UCLA David Geffen School of Medicine, Los Angeles. Genotyping was )] TJ ET BT 26.250 335.682 Td /F1 9.8 Tf [(completed by Laragen \(Los Angeles, CA, USA\). BACHD and the corresponding WT control mice were tested at 16, 24, 36, and )] TJ ET BT 26.250 323.777 Td /F1 9.8 Tf [(52 weeks of age.)] TJ ET BT 26.250 304.372 Td /F1 9.8 Tf [(Prior to testing in PhenoCube)] TJ ET BT 153.605 308.260 Td /F1 8.7 Tf [(TM)] TJ ET BT 166.119 304.372 Td /F1 9.8 Tf [(, mice were injected with sterile transponders \(T-IS 8010 FDX-B, Datamars SA, Bedano, )] TJ ET BT 26.250 292.467 Td /F1 9.8 Tf [(Switzerland\) under 2% isoflurane inhalation anesthesia. During recovery from anesthesia, mice were introduced to their social )] TJ ET BT 26.250 280.563 Td /F1 9.8 Tf [(testing groups and housed in genetically homogenous in OptiRAT cages \(Animal Care Systems, CO\). The environment was )] TJ ET BT 26.250 268.658 Td /F1 9.8 Tf [(enriched with a play tunnel, shredded paper, and a plastic bone. Food and water were available ad libitum with the exception of )] TJ ET BT 26.250 256.753 Td /F1 9.8 Tf [(water deprivation 16 hours prior to each phase of testing in our system. Temperature and humidity were controlled and )] TJ ET BT 26.250 244.848 Td /F1 9.8 Tf [(monitored daily. This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of )] TJ ET BT 26.250 232.944 Td /F1 9.8 Tf [(Laboratory Animals, NRC )] TJ ET 0.267 0.267 0.267 rg BT 139.487 234.451 Td /F4 8.7 Tf [(23)] TJ ET 0.271 0.267 0.267 rg BT 149.124 232.944 Td /F1 9.8 Tf [(. The protocol was approved by the Institutional Animal Care and Use Committee of Psychogenics, )] TJ ET BT 26.250 221.039 Td /F1 9.8 Tf [(Inc. \(PHS OLAW animal welfare assurance number A4471-01\), an AAALAC International accredited institution \(Unit #001213\).)] TJ ET BT 26.250 201.634 Td /F4 9.8 Tf [(Apparatus:)] TJ ET BT 26.250 182.229 Td /F1 9.8 Tf [(Experiments were conducted using modified Intellicage units \(New Behavior, AG, Zurich, CH\), each with a Day/Night Camera )] TJ ET BT 26.250 170.325 Td /F1 9.8 Tf [(mounted on top of the cage for Computer Vision \(CV\) analysis \(Figure 1\). Intra-maze spatial cues were added to the )] TJ ET BT 26.250 158.420 Td /F1 9.8 Tf [(environment by placing laminated striped paper on the outside of the cage walls, while three climbing structures \(two rods, a )] TJ ET BT 26.250 146.515 Td /F1 9.8 Tf [(cubic central object and a three step staircase\) were placed inside the cage to provide an enriched topology \(Figure 1 left )] TJ ET BT 26.250 134.610 Td /F1 9.8 Tf [(panel\).)] TJ ET BT 26.250 115.206 Td /F1 9.8 Tf [(The cages were maintained at all times on a 12:12 light/dark cycle, with white light during the day and red light during the night. )] TJ ET BT 26.250 103.301 Td /F1 9.8 Tf [(The light intensity under red light was recorded at 7 lux using a photographic band-pass filter \(LDP LLC, NJ\) that eliminates long )] TJ ET BT 26.250 91.396 Td /F1 9.8 Tf [(wavelength light frequencies not visible to mice )] TJ ET 0.267 0.267 0.267 rg BT 232.170 92.903 Td /F4 8.7 Tf [(24)] TJ ET 0.271 0.267 0.267 rg BT 241.807 91.396 Td /F1 9.8 Tf [( and was sufficient to allow the camera to detect the mice while maintaining a )] TJ ET BT 26.250 79.491 Td /F1 9.8 Tf [(low subjective light level for the mice. The light intensity under white light was 100 lux. Inside the PhenoCube)] TJ ET BT 495.001 83.380 Td /F1 8.7 Tf [(TM)] TJ ET BT 507.515 79.491 Td /F1 9.8 Tf [( environment, )] TJ ET BT 26.250 67.587 Td /F1 9.8 Tf [(water was only available from within the corners while food was freely available on the cage floor at all times. It was sometimes )] TJ ET BT 26.250 55.682 Td /F1 9.8 Tf [(necessary to remove a mouse from the study when no licking was recorded, in order to ensure proper hydration.)] TJ ET Q q 15.000 45.801 577.500 731.199 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(mean efficient and quick screening of animal subjects for multiple phenotypes simultaneously. These phenotypes might include )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(\(but are not limited to\) cognitive performance, motivation and locomotor, circadian, and exploratory activity. In addition, we )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(showed that the system has high sensitivity and robustness to detect the deficits in these mutant lines.)] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(The R6/2 mouse model expresses a short N-terminal fragment of mutant human huntingtin and is the best-known mouse model )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(of HD )] TJ ET 0.267 0.267 0.267 rg BT 53.882 713.864 Td /F4 8.7 Tf [(5)] TJ ET 0.271 0.267 0.267 rg BT 58.700 712.357 Td /F1 9.8 Tf [( in terms of pathophysiology and behavioral abnormalities \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 335.074 713.864 Td /F4 8.7 Tf [(7)] TJ ET 0.271 0.267 0.267 rg BT 339.892 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 342.302 713.864 Td /F4 8.7 Tf [(8)] TJ ET 0.271 0.267 0.267 rg BT 347.120 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 349.530 713.864 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 354.348 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 356.758 713.864 Td /F4 8.7 Tf [(10)] TJ ET 0.271 0.267 0.267 rg BT 366.395 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 368.804 713.864 Td /F4 8.7 Tf [(11)] TJ ET 0.271 0.267 0.267 rg BT 378.442 712.357 Td /F1 9.8 Tf [(\). R6/2 mice have been shown to be )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(hypoactive \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 100.496 701.960 Td /F4 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 110.134 704.341 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 112.543 701.960 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 117.362 700.452 Td /F1 9.8 Tf [(\), exhibit disrupted circadian patterns \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 304.318 701.960 Td /F4 8.7 Tf [(13)] TJ ET 0.271 0.267 0.267 rg BT 313.955 700.452 Td /F1 9.8 Tf [(\), response acquisition )] TJ ET 0.267 0.267 0.267 rg BT 414.205 701.960 Td /F4 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 423.842 700.452 Td /F1 9.8 Tf [( and interval timing )] TJ ET 0.267 0.267 0.267 rg BT 508.375 701.960 Td /F4 8.7 Tf [(15)] TJ ET 0.271 0.267 0.267 rg BT 518.012 700.452 Td /F1 9.8 Tf [(, and have )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(impaired spatial discrimination and reversal as assessed in water-maze and t-maze tasks \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 438.597 690.055 Td /F4 8.7 Tf [(8)] TJ ET 0.271 0.267 0.267 rg BT 443.416 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 445.825 690.055 Td /F4 8.7 Tf [(16)] TJ ET 0.271 0.267 0.267 rg BT 455.462 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 457.872 690.055 Td /F4 8.7 Tf [(17)] TJ ET 0.271 0.267 0.267 rg BT 467.509 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 469.918 690.055 Td /F4 8.7 Tf [(18)] TJ ET 0.271 0.267 0.267 rg BT 479.556 692.436 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 481.965 690.055 Td /F4 8.7 Tf [(19)] TJ ET 0.271 0.267 0.267 rg BT 491.602 688.548 Td /F1 9.8 Tf [(\). By contrast, the )] TJ ET BT 26.250 676.643 Td /F1 9.8 Tf [(BACHD mouse expresses the full-length human mutant huntingtin gene with 97 glutamine repeats under the control of )] TJ ET BT 26.250 664.738 Td /F1 9.8 Tf [(endogenous htt regulatory machinery on a bacterial artificial chromosome, and has been less widely studied but also has been )] TJ ET BT 26.250 652.833 Td /F1 9.8 Tf [(shown to mimic some aspects of HD pathophysiology and behavioral abnormalities )] TJ ET 0.267 0.267 0.267 rg BT 387.137 654.341 Td /F4 8.7 Tf [(4)] TJ ET 0.271 0.267 0.267 rg BT 391.955 656.722 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 394.365 654.341 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 399.183 656.722 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 401.593 654.341 Td /F4 8.7 Tf [(20)] TJ ET 0.271 0.267 0.267 rg BT 411.230 652.833 Td /F1 9.8 Tf [(, including circadian abnormalities )] TJ ET 0.267 0.267 0.267 rg BT 560.239 654.341 Td /F4 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 569.876 656.722 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 26.250 642.436 Td /F4 8.7 Tf [(21)] TJ ET 0.271 0.267 0.267 rg BT 35.887 640.929 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(The PhenoCube)] TJ ET BT 97.249 625.412 Td /F1 8.7 Tf [(TM)] TJ ET BT 109.764 621.524 Td /F1 9.8 Tf [( system acquires a broad range of different measures that span multiple disease-relevant domains \(i.e., )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(cognition, locomotor activity and circadian patterns\), and thus can efficiently capture the complexity of the behavioral phenotype. )] TJ ET BT 26.250 597.714 Td /F1 9.8 Tf [(We developed the PhenoCube)] TJ ET BT 159.035 601.603 Td /F1 8.7 Tf [(TM)] TJ ET BT 171.550 597.714 Td /F1 9.8 Tf [( through hardware modifications of IntelliCage units and addition of custom-built computer )] TJ ET BT 26.250 585.810 Td /F1 9.8 Tf [(vision hardware and software. This system enables behavioral phenotyping of group-housed mice within a home-cage-like )] TJ ET BT 26.250 573.905 Td /F1 9.8 Tf [(environment over multiple days with minimal experimenter interruption, allowing capture of the endogenous behavioral rhythms )] TJ ET BT 26.250 562.000 Td /F1 9.8 Tf [(of the subjects. Models of HD are particularly relevant disease model for demonstration of the utility of this system as it is )] TJ ET BT 26.250 550.095 Td /F1 9.8 Tf [(characterized by altered cognition, motor activity, and circadian rhythms, all domains that can all be assessed simultaneously by )] TJ ET BT 26.250 538.191 Td /F1 9.8 Tf [(the PhenoCube)] TJ ET BT 94.003 542.079 Td /F1 8.7 Tf [(TM)] TJ ET BT 106.517 538.191 Td /F1 9.8 Tf [(. The present experiments capture these behavioral phenotypes using the novel PhenoCube)] TJ ET BT 504.854 542.079 Td /F1 8.7 Tf [(TM)] TJ ET BT 517.368 538.191 Td /F1 9.8 Tf [( apparatus, )] TJ ET BT 26.250 526.286 Td /F1 9.8 Tf [(demonstrating the utility of the system and of the experimental protocols developed in our lab.)] TJ ET BT 26.250 489.683 Td /F4 12.0 Tf [(METHODS)] TJ ET BT 26.250 469.729 Td /F4 9.8 Tf [(Animals:)] TJ ET BT 26.250 450.324 Td /F1 9.8 Tf [(R6/2 \(R6/2 CAG 120, CHDI-81001000\) transgenic heterozygous and wild type \(WT\) littermate control mice were obtained from )] TJ ET BT 26.250 438.420 Td /F1 9.8 Tf [(Jackson Laboratories \(Bar Harbor,ME; strain B6CBA-Tg[HDexon1]62Gpb/3J, stock number 006494\). Mice were generated by )] TJ ET BT 26.250 426.515 Td /F1 9.8 Tf [(crossing ovarian-transplanted females \(from R6/2 CBAB6J female donors\) with CBAB6F1J WT males. Genotype was )] TJ ET BT 26.250 414.610 Td /F1 9.8 Tf [(determined by polymerase chain reaction \(PCR\) of tail-tip DNA at 15 days of age )] TJ ET 0.267 0.267 0.267 rg BT 377.387 416.117 Td /F4 8.7 Tf [(22)] TJ ET 0.271 0.267 0.267 rg BT 387.024 414.610 Td /F1 9.8 Tf [(. CAG repeat lengths were measured by )] TJ ET BT 26.250 402.705 Td /F1 9.8 Tf [(Laragen \(Los Angeles, CA, USA\) using standard protocols and Genemapper software as previously described )] TJ ET 0.267 0.267 0.267 rg BT 503.668 404.213 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 508.487 402.705 Td /F1 9.8 Tf [(. R6/2 mice )] TJ ET BT 26.250 390.801 Td /F1 9.8 Tf [(were tested at 7, 8, 9 and 10 weeks of age. The CAG repeat of the R6/2 mutant is 120 )] TJ ET BT 400.757 390.801 Td /F1 9.8 Tf [(+)] TJ ET BT 406.451 390.801 Td /F1 9.8 Tf [( 5.)] TJ ET BT 26.250 371.396 Td /F1 9.8 Tf [(BACHD \(CHDI-81001012and WT control mice were generated at the Jackson laboratory on an FVB/NJ x C57Bl/6J F1, created )] TJ ET BT 26.250 359.491 Td /F1 9.8 Tf [(by crossing BAC transgenic FVB/NJ males with WT C57Bl/6J females. The congenic BAC FVB/NJ line used for breeding was )] TJ ET BT 26.250 347.586 Td /F1 9.8 Tf [(obtained from the laboratory of X. William Yang at the UCLA David Geffen School of Medicine, Los Angeles. Genotyping was )] TJ ET BT 26.250 335.682 Td /F1 9.8 Tf [(completed by Laragen \(Los Angeles, CA, USA\). BACHD and the corresponding WT control mice were tested at 16, 24, 36, and )] TJ ET BT 26.250 323.777 Td /F1 9.8 Tf [(52 weeks of age.)] TJ ET BT 26.250 304.372 Td /F1 9.8 Tf [(Prior to testing in PhenoCube)] TJ ET BT 153.605 308.260 Td /F1 8.7 Tf [(TM)] TJ ET BT 166.119 304.372 Td /F1 9.8 Tf [(, mice were injected with sterile transponders \(T-IS 8010 FDX-B, Datamars SA, Bedano, )] TJ ET BT 26.250 292.467 Td /F1 9.8 Tf [(Switzerland\) under 2% isoflurane inhalation anesthesia. During recovery from anesthesia, mice were introduced to their social )] TJ ET BT 26.250 280.563 Td /F1 9.8 Tf [(testing groups and housed in genetically homogenous in OptiRAT cages \(Animal Care Systems, CO\). The environment was )] TJ ET BT 26.250 268.658 Td /F1 9.8 Tf [(enriched with a play tunnel, shredded paper, and a plastic bone. Food and water were available ad libitum with the exception of )] TJ ET BT 26.250 256.753 Td /F1 9.8 Tf [(water deprivation 16 hours prior to each phase of testing in our system. Temperature and humidity were controlled and )] TJ ET BT 26.250 244.848 Td /F1 9.8 Tf [(monitored daily. This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of )] TJ ET BT 26.250 232.944 Td /F1 9.8 Tf [(Laboratory Animals, NRC )] TJ ET 0.267 0.267 0.267 rg BT 139.487 234.451 Td /F4 8.7 Tf [(23)] TJ ET 0.271 0.267 0.267 rg BT 149.124 232.944 Td /F1 9.8 Tf [(. The protocol was approved by the Institutional Animal Care and Use Committee of Psychogenics, )] TJ ET BT 26.250 221.039 Td /F1 9.8 Tf [(Inc. \(PHS OLAW animal welfare assurance number A4471-01\), an AAALAC International accredited institution \(Unit #001213\).)] TJ ET BT 26.250 201.634 Td /F4 9.8 Tf [(Apparatus:)] TJ ET BT 26.250 182.229 Td /F1 9.8 Tf [(Experiments were conducted using modified Intellicage units \(New Behavior, AG, Zurich, CH\), each with a Day/Night Camera )] TJ ET BT 26.250 170.325 Td /F1 9.8 Tf [(mounted on top of the cage for Computer Vision \(CV\) analysis \(Figure 1\). Intra-maze spatial cues were added to the )] TJ ET BT 26.250 158.420 Td /F1 9.8 Tf [(environment by placing laminated striped paper on the outside of the cage walls, while three climbing structures \(two rods, a )] TJ ET BT 26.250 146.515 Td /F1 9.8 Tf [(cubic central object and a three step staircase\) were placed inside the cage to provide an enriched topology \(Figure 1 left )] TJ ET BT 26.250 134.610 Td /F1 9.8 Tf [(panel\).)] TJ ET BT 26.250 115.206 Td /F1 9.8 Tf [(The cages were maintained at all times on a 12:12 light/dark cycle, with white light during the day and red light during the night. )] TJ ET BT 26.250 103.301 Td /F1 9.8 Tf [(The light intensity under red light was recorded at 7 lux using a photographic band-pass filter \(LDP LLC, NJ\) that eliminates long )] TJ ET BT 26.250 91.396 Td /F1 9.8 Tf [(wavelength light frequencies not visible to mice )] TJ ET 0.267 0.267 0.267 rg BT 232.170 92.903 Td /F4 8.7 Tf [(24)] TJ ET 0.271 0.267 0.267 rg BT 241.807 91.396 Td /F1 9.8 Tf [( and was sufficient to allow the camera to detect the mice while maintaining a )] TJ ET BT 26.250 79.491 Td /F1 9.8 Tf [(low subjective light level for the mice. The light intensity under white light was 100 lux. Inside the PhenoCube)] TJ ET BT 495.001 83.380 Td /F1 8.7 Tf [(TM)] TJ ET BT 507.515 79.491 Td /F1 9.8 Tf [( environment, )] TJ ET BT 26.250 67.587 Td /F1 9.8 Tf [(water was only available from within the corners while food was freely available on the cage floor at all times. It was sometimes )] TJ ET BT 26.250 55.682 Td /F1 9.8 Tf [(necessary to remove a mouse from the study when no licking was recorded, in order to ensure proper hydration.)] TJ ET Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(2)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Huntington Disease)] TJ ET Q endstream endobj 148 0 obj << /Type /Annot /Subtype /Link /A 149 0 R /Border [0 0 0] /H /I /Rect [ 53.8815 713.0627 58.7002 721.8810 ] >> endobj 149 0 obj << /Type /Action >> endobj 150 0 obj << /Type /Annot /Subtype /Link /A 151 0 R /Border [0 0 0] /H /I /Rect [ 335.0737 713.0627 339.8923 721.8810 ] >> endobj 151 0 obj << /Type /Action >> endobj 152 0 obj << /Type /Annot /Subtype /Link /A 153 0 R /Border [0 0 0] /H /I /Rect [ 342.3017 713.0627 347.1203 721.8810 ] >> endobj 153 0 obj << /Type /Action >> endobj 154 0 obj << /Type /Annot /Subtype /Link /A 155 0 R /Border [0 0 0] /H /I /Rect [ 349.5297 713.0627 354.3483 721.8810 ] >> endobj 155 0 obj << /Type /Action >> 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>> stream 0.271 0.267 0.267 rg q 15.000 40.145 577.500 736.855 re W n 0.965 0.965 0.965 rg 26.250 628.359 555.000 148.641 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 777.000 m 581.250 777.000 l 581.250 776.250 l 26.250 776.250 l f 26.250 628.359 m 581.250 628.359 l 581.250 629.109 l 26.250 629.109 l f q 225.000 0 0 82.500 35.250 684.750 cm /I3 Do Q q 35.250 639.609 537.000 39.141 re W n 0.271 0.267 0.267 rg BT 35.250 669.226 Td /F4 9.8 Tf [(Fig. 1: PhenoCube)] TJ ET BT 35.250 649.856 Td /F1 9.8 Tf [(Inside \(left panel\) and outside view \(right panel\).)] TJ ET Q BT 26.250 611.335 Td /F4 9.8 Tf [(Procedure:)] TJ ET BT 26.250 591.930 Td /F5 9.8 Tf [(Standard Experimental Procedure:)] TJ ET BT 175.815 591.930 Td /F1 9.8 Tf [(In all the experiments described here, a standard series of test protocols were employed, )] TJ ET BT 26.250 580.026 Td /F1 9.8 Tf [(defined here as Habituation, Alternation, and Reversal \(see Figure 2\).)] TJ ET BT 26.250 560.621 Td /F5 9.8 Tf [(Habituation: )] TJ ET BT 80.987 560.621 Td /F1 9.8 Tf [(The Habituation phase \(Figure 2 left panel\) was used at the start of the experiment during which mice have )] TJ ET BT 26.250 548.716 Td /F1 9.8 Tf [(access to all four of the corners. As soon as any mouse enters, both doors to the water open and remain open until the mouse )] TJ ET BT 26.250 536.811 Td /F1 9.8 Tf [(leaves the corner. This phase of the experiment constitutes the magazine training and allows the mice to learn that water )] TJ ET BT 26.250 524.907 Td /F1 9.8 Tf [(reinforcement is available in the corners. Basic activity data is collected during this phase of the experiments. The overall rate of )] TJ ET BT 26.250 513.002 Td /F1 9.8 Tf [(corner visits and licking provide an insight into the general activity levels of the animals as well as their capacity to obtain water )] TJ ET BT 26.250 501.097 Td /F1 9.8 Tf [(reinforcement, while evaluation of the overall rate of repeated visits \(i.e. returning to the previously visited corner\) gives some )] TJ ET BT 26.250 489.192 Td /F1 9.8 Tf [(indication of the animals general tendency to persevere.)] TJ ET BT 26.250 469.788 Td /F5 9.8 Tf [(Alternation: )] TJ ET BT 78.276 469.788 Td /F1 9.8 Tf [(In the Alternation training protocol \(Figure 2 middle panel\), the animals are required to visit two of the four corners )] TJ ET BT 26.250 457.883 Td /F1 9.8 Tf [(in order to gain access to water. For each subject, two adjacent corners \(active corners\) along one of the shorter sides of the )] TJ ET BT 26.250 445.978 Td /F1 9.8 Tf [(rectangular cage were contingently rewarded, while the other two \(exploratory corners\) were never rewarded. The Alternation )] TJ ET BT 26.250 434.073 Td /F1 9.8 Tf [(protocol was set up to train the animals to switch between the two active corners, only receiving reinforcement for alternating )] TJ ET BT 26.250 422.169 Td /F1 9.8 Tf [(visits. For example, if corners 1 and 2 were active, an initial visit to corner 1 would be a correct visit and rewarded. To obtain )] TJ ET BT 26.250 410.264 Td /F1 9.8 Tf [(further reinforcement, the mouse was then required to visit corner 2. Repeat visits to corner 1 were classified as incorrect, and )] TJ ET BT 26.250 398.359 Td /F1 9.8 Tf [(mice would not receive a reward there. Following a visit to corner 2, the corners would switch again, such that reinforcement )] TJ ET BT 26.250 386.454 Td /F1 9.8 Tf [(would now be available only in corner 1 and so on. If a mouse incorrectly visits the same active corner more than once or visits )] TJ ET BT 26.250 374.550 Td /F1 9.8 Tf [(an exploratory corner after leaving an active corner, the identity of the target correct corner does not change and a visit to that )] TJ ET BT 26.250 362.645 Td /F1 9.8 Tf [(corner will result in available reinforcement; the only event leading to a switch in the correct corner identity is a visit to the )] TJ ET BT 26.250 350.740 Td /F1 9.8 Tf [(currently correct corner, in which reinforcement will be available.)] TJ ET BT 26.250 331.335 Td /F1 9.8 Tf [(Alternation data was calculated within a 113-s interval of leaving an active corner, such that only a visit to the correct corner 113-)] TJ ET BT 26.250 319.431 Td /F1 9.8 Tf [(s or less after exiting an active corner counted as a correct Alternation, with any visit to the currently )] TJ ET BT 458.682 319.431 Td /F5 9.8 Tf [(incorrect corner)] TJ ET BT 526.406 319.431 Td /F1 9.8 Tf [( counting )] TJ ET BT 26.250 307.526 Td /F1 9.8 Tf [(as incorrect, while visits to the )] TJ ET BT 158.460 307.526 Td /F5 9.8 Tf [(exploratory corners)] TJ ET BT 241.901 307.526 Td /F1 9.8 Tf [( are irrelevant. Inter visit intervals were cap to minimize the chances that other )] TJ ET BT 26.250 295.621 Td /F1 9.8 Tf [(events interrupted the alternating sequence the precise interval used is not of great significance, with similar results detected )] TJ ET BT 26.250 283.716 Td /F1 9.8 Tf [(with different intervals. Irrespective of this interval, visits to a correct corner were rewarded the interval is relevant only to the )] TJ ET BT 26.250 271.812 Td /F1 9.8 Tf [(data handling process. The primary measure employed to assess Alternation performance, percent alternation, is generated by )] TJ ET BT 26.250 259.907 Td /F1 9.8 Tf [(calculating the percentage of visits to active corners made following a visit to either of the two active corners within a particular )] TJ ET BT 26.250 248.002 Td /F1 9.8 Tf [(interval that are correct alternation visits. For example, if corners 1 and 2 are active, the number of visits to corner 2 following a )] TJ ET BT 26.250 236.097 Td /F1 9.8 Tf [(visit to corner 1 plus the number of visits to corner 1 following a visit to corner 2 comprises the total Alternation visits, which are )] TJ ET BT 26.250 224.193 Td /F1 9.8 Tf [(then divided by the total visits to either corner 1 or corner 2. Previous experiments in our laboratories have indicated that, )] TJ ET BT 26.250 212.288 Td /F1 9.8 Tf [(although mice do have some tendency to spontaneously alternate between foraging locations, their alternating behavior within )] TJ ET BT 26.250 200.383 Td /F1 9.8 Tf [(the corners is significantly increased by the type of enforced alternation training employed here, such that performance on this )] TJ ET BT 26.250 188.478 Td /F1 9.8 Tf [(task does appear to be sensitive to the animals learning the reinforcement contingencies.)] TJ ET BT 26.250 169.074 Td /F1 9.8 Tf [(Each corner contained two nosepoke recesses, one on each side, used to deliver water reinforcement during correct visits. )] TJ ET BT 26.250 157.169 Td /F1 9.8 Tf [(Each active corner delivered water only on one side, with the rewarded side dependant on the specific identity of the current )] TJ ET BT 26.250 145.264 Td /F1 9.8 Tf [(corner, such that if only the left side was rewarded in active corner 1, then only the right side was rewarded in active corner 2 )] TJ ET BT 26.250 133.359 Td /F1 9.8 Tf [(and vice versa. When mice made visits to a correct corner \(i.e., an alternation visit\) and nose-poked on the correct side, the )] TJ ET BT 26.250 121.455 Td /F1 9.8 Tf [(door behind the correct nosepoke recess for that corner opened for 8 s, providing access to water. After 8 s, the door closed, )] TJ ET BT 26.250 109.550 Td /F1 9.8 Tf [(preventing further access to water, with the mice then required to alternate and enter the other active corner for further rewards. )] TJ ET BT 26.250 97.645 Td /F1 9.8 Tf [(No penalty was imposed for initially nosepoking on the incorrect side; the first nosepoke during a correct visit to the correct side )] TJ ET BT 26.250 85.740 Td /F1 9.8 Tf [(always resulted in delivery of reinforcement. However, for the purposes of behavioral analysis, the proportion of initial )] TJ ET BT 26.250 73.836 Td /F1 9.8 Tf [(nosepokes to the correct side was calculated such that, for example, a mouse making a correct visit and subsequently )] TJ ET BT 26.250 61.931 Td /F1 9.8 Tf [(nosepoking to the wrong side and then to the correct side would receive water, but the mouses initial nosepoke for that visit )] TJ ET BT 26.250 50.026 Td /F1 9.8 Tf [(would be scored as incorrect \(from a behavioral efficiency perspective\).)] TJ ET Q q 15.000 40.145 577.500 736.855 re W n 0.965 0.965 0.965 rg 26.250 628.359 555.000 148.641 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 777.000 m 581.250 777.000 l 581.250 776.250 l 26.250 776.250 l f 26.250 628.359 m 581.250 628.359 l 581.250 629.109 l 26.250 629.109 l f q 225.000 0 0 82.500 35.250 684.750 cm /I3 Do Q q 35.250 639.609 537.000 39.141 re W n 0.271 0.267 0.267 rg BT 35.250 669.226 Td /F4 9.8 Tf [(Fig. 1: PhenoCube)] TJ ET BT 35.250 649.856 Td /F1 9.8 Tf [(Inside \(left panel\) and outside view \(right panel\).)] TJ ET Q BT 26.250 611.335 Td /F4 9.8 Tf [(Procedure:)] TJ ET BT 26.250 591.930 Td /F5 9.8 Tf [(Standard Experimental Procedure:)] TJ ET BT 175.815 591.930 Td /F1 9.8 Tf [(In all the experiments described here, a standard series of test protocols were employed, )] TJ ET BT 26.250 580.026 Td /F1 9.8 Tf [(defined here as Habituation, Alternation, and Reversal \(see Figure 2\).)] TJ ET BT 26.250 560.621 Td /F5 9.8 Tf [(Habituation: )] TJ ET BT 80.987 560.621 Td /F1 9.8 Tf [(The Habituation phase \(Figure 2 left panel\) was used at the start of the experiment during which mice have )] TJ ET BT 26.250 548.716 Td /F1 9.8 Tf [(access to all four of the corners. As soon as any mouse enters, both doors to the water open and remain open until the mouse )] TJ ET BT 26.250 536.811 Td /F1 9.8 Tf [(leaves the corner. This phase of the experiment constitutes the magazine training and allows the mice to learn that water )] TJ ET BT 26.250 524.907 Td /F1 9.8 Tf [(reinforcement is available in the corners. Basic activity data is collected during this phase of the experiments. The overall rate of )] TJ ET BT 26.250 513.002 Td /F1 9.8 Tf [(corner visits and licking provide an insight into the general activity levels of the animals as well as their capacity to obtain water )] TJ ET BT 26.250 501.097 Td /F1 9.8 Tf [(reinforcement, while evaluation of the overall rate of repeated visits \(i.e. returning to the previously visited corner\) gives some )] TJ ET BT 26.250 489.192 Td /F1 9.8 Tf [(indication of the animals general tendency to persevere.)] TJ ET BT 26.250 469.788 Td /F5 9.8 Tf [(Alternation: )] TJ ET BT 78.276 469.788 Td /F1 9.8 Tf [(In the Alternation training protocol \(Figure 2 middle panel\), the animals are required to visit two of the four corners )] TJ ET BT 26.250 457.883 Td /F1 9.8 Tf [(in order to gain access to water. For each subject, two adjacent corners \(active corners\) along one of the shorter sides of the )] TJ ET BT 26.250 445.978 Td /F1 9.8 Tf [(rectangular cage were contingently rewarded, while the other two \(exploratory corners\) were never rewarded. The Alternation )] TJ ET BT 26.250 434.073 Td /F1 9.8 Tf [(protocol was set up to train the animals to switch between the two active corners, only receiving reinforcement for alternating )] TJ ET BT 26.250 422.169 Td /F1 9.8 Tf [(visits. For example, if corners 1 and 2 were active, an initial visit to corner 1 would be a correct visit and rewarded. To obtain )] TJ ET BT 26.250 410.264 Td /F1 9.8 Tf [(further reinforcement, the mouse was then required to visit corner 2. Repeat visits to corner 1 were classified as incorrect, and )] TJ ET BT 26.250 398.359 Td /F1 9.8 Tf [(mice would not receive a reward there. Following a visit to corner 2, the corners would switch again, such that reinforcement )] TJ ET BT 26.250 386.454 Td /F1 9.8 Tf [(would now be available only in corner 1 and so on. If a mouse incorrectly visits the same active corner more than once or visits )] TJ ET BT 26.250 374.550 Td /F1 9.8 Tf [(an exploratory corner after leaving an active corner, the identity of the target correct corner does not change and a visit to that )] TJ ET BT 26.250 362.645 Td /F1 9.8 Tf [(corner will result in available reinforcement; the only event leading to a switch in the correct corner identity is a visit to the )] TJ ET BT 26.250 350.740 Td /F1 9.8 Tf [(currently correct corner, in which reinforcement will be available.)] TJ ET BT 26.250 331.335 Td /F1 9.8 Tf [(Alternation data was calculated within a 113-s interval of leaving an active corner, such that only a visit to the correct corner 113-)] TJ ET BT 26.250 319.431 Td /F1 9.8 Tf [(s or less after exiting an active corner counted as a correct Alternation, with any visit to the currently )] TJ ET BT 458.682 319.431 Td /F5 9.8 Tf [(incorrect corner)] TJ ET BT 526.406 319.431 Td /F1 9.8 Tf [( counting )] TJ ET BT 26.250 307.526 Td /F1 9.8 Tf [(as incorrect, while visits to the )] TJ ET BT 158.460 307.526 Td /F5 9.8 Tf [(exploratory corners)] TJ ET BT 241.901 307.526 Td /F1 9.8 Tf [( are irrelevant. Inter visit intervals were cap to minimize the chances that other )] TJ ET BT 26.250 295.621 Td /F1 9.8 Tf [(events interrupted the alternating sequence the precise interval used is not of great significance, with similar results detected )] TJ ET BT 26.250 283.716 Td /F1 9.8 Tf [(with different intervals. Irrespective of this interval, visits to a correct corner were rewarded the interval is relevant only to the )] TJ ET BT 26.250 271.812 Td /F1 9.8 Tf [(data handling process. The primary measure employed to assess Alternation performance, percent alternation, is generated by )] TJ ET BT 26.250 259.907 Td /F1 9.8 Tf [(calculating the percentage of visits to active corners made following a visit to either of the two active corners within a particular )] TJ ET BT 26.250 248.002 Td /F1 9.8 Tf [(interval that are correct alternation visits. For example, if corners 1 and 2 are active, the number of visits to corner 2 following a )] TJ ET BT 26.250 236.097 Td /F1 9.8 Tf [(visit to corner 1 plus the number of visits to corner 1 following a visit to corner 2 comprises the total Alternation visits, which are )] TJ ET BT 26.250 224.193 Td /F1 9.8 Tf [(then divided by the total visits to either corner 1 or corner 2. Previous experiments in our laboratories have indicated that, )] TJ ET BT 26.250 212.288 Td /F1 9.8 Tf [(although mice do have some tendency to spontaneously alternate between foraging locations, their alternating behavior within )] TJ ET BT 26.250 200.383 Td /F1 9.8 Tf [(the corners is significantly increased by the type of enforced alternation training employed here, such that performance on this )] TJ ET BT 26.250 188.478 Td /F1 9.8 Tf [(task does appear to be sensitive to the animals learning the reinforcement contingencies.)] TJ ET BT 26.250 169.074 Td /F1 9.8 Tf [(Each corner contained two nosepoke recesses, one on each side, used to deliver water reinforcement during correct visits. )] TJ ET BT 26.250 157.169 Td /F1 9.8 Tf [(Each active corner delivered water only on one side, with the rewarded side dependant on the specific identity of the current )] TJ ET BT 26.250 145.264 Td /F1 9.8 Tf [(corner, such that if only the left side was rewarded in active corner 1, then only the right side was rewarded in active corner 2 )] TJ ET BT 26.250 133.359 Td /F1 9.8 Tf [(and vice versa. When mice made visits to a correct corner \(i.e., an alternation visit\) and nose-poked on the correct side, the )] TJ ET BT 26.250 121.455 Td /F1 9.8 Tf [(door behind the correct nosepoke recess for that corner opened for 8 s, providing access to water. After 8 s, the door closed, )] TJ ET BT 26.250 109.550 Td /F1 9.8 Tf [(preventing further access to water, with the mice then required to alternate and enter the other active corner for further rewards. )] TJ ET BT 26.250 97.645 Td /F1 9.8 Tf [(No penalty was imposed for initially nosepoking on the incorrect side; the first nosepoke during a correct visit to the correct side )] TJ ET BT 26.250 85.740 Td /F1 9.8 Tf [(always resulted in delivery of reinforcement. However, for the purposes of behavioral analysis, the proportion of initial )] TJ ET BT 26.250 73.836 Td /F1 9.8 Tf [(nosepokes to the correct side was calculated such that, for example, a mouse making a correct visit and subsequently )] TJ ET BT 26.250 61.931 Td /F1 9.8 Tf [(nosepoking to the wrong side and then to the correct side would receive water, but the mouses initial nosepoke for that visit )] TJ ET BT 26.250 50.026 Td /F1 9.8 Tf [(would be scored as incorrect \(from a behavioral efficiency perspective\).)] TJ ET Q q 15.000 40.145 577.500 736.855 re W n 0.965 0.965 0.965 rg 26.250 628.359 555.000 148.641 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 777.000 m 581.250 777.000 l 581.250 776.250 l 26.250 776.250 l f 26.250 628.359 m 581.250 628.359 l 581.250 629.109 l 26.250 629.109 l f q 225.000 0 0 82.500 35.250 684.750 cm /I3 Do Q q 35.250 639.609 537.000 39.141 re W n 0.271 0.267 0.267 rg BT 35.250 669.226 Td /F4 9.8 Tf [(Fig. 1: PhenoCube)] TJ ET BT 35.250 649.856 Td /F1 9.8 Tf [(Inside \(left panel\) and outside view \(right panel\).)] TJ ET Q BT 26.250 611.335 Td /F4 9.8 Tf [(Procedure:)] TJ ET BT 26.250 591.930 Td /F5 9.8 Tf [(Standard Experimental Procedure:)] TJ ET BT 175.815 591.930 Td /F1 9.8 Tf [(In all the experiments described here, a standard series of test protocols were employed, )] TJ ET BT 26.250 580.026 Td /F1 9.8 Tf [(defined here as Habituation, Alternation, and Reversal \(see Figure 2\).)] TJ ET BT 26.250 560.621 Td /F5 9.8 Tf [(Habituation: )] TJ ET BT 80.987 560.621 Td /F1 9.8 Tf [(The Habituation phase \(Figure 2 left panel\) was used at the start of the experiment during which mice have )] TJ ET BT 26.250 548.716 Td /F1 9.8 Tf [(access to all four of the corners. As soon as any mouse enters, both doors to the water open and remain open until the mouse )] TJ ET BT 26.250 536.811 Td /F1 9.8 Tf [(leaves the corner. This phase of the experiment constitutes the magazine training and allows the mice to learn that water )] TJ ET BT 26.250 524.907 Td /F1 9.8 Tf [(reinforcement is available in the corners. Basic activity data is collected during this phase of the experiments. The overall rate of )] TJ ET BT 26.250 513.002 Td /F1 9.8 Tf [(corner visits and licking provide an insight into the general activity levels of the animals as well as their capacity to obtain water )] TJ ET BT 26.250 501.097 Td /F1 9.8 Tf [(reinforcement, while evaluation of the overall rate of repeated visits \(i.e. returning to the previously visited corner\) gives some )] TJ ET BT 26.250 489.192 Td /F1 9.8 Tf [(indication of the animals general tendency to persevere.)] TJ ET BT 26.250 469.788 Td /F5 9.8 Tf [(Alternation: )] TJ ET BT 78.276 469.788 Td /F1 9.8 Tf [(In the Alternation training protocol \(Figure 2 middle panel\), the animals are required to visit two of the four corners )] TJ ET BT 26.250 457.883 Td /F1 9.8 Tf [(in order to gain access to water. For each subject, two adjacent corners \(active corners\) along one of the shorter sides of the )] TJ ET BT 26.250 445.978 Td /F1 9.8 Tf [(rectangular cage were contingently rewarded, while the other two \(exploratory corners\) were never rewarded. The Alternation )] TJ ET BT 26.250 434.073 Td /F1 9.8 Tf [(protocol was set up to train the animals to switch between the two active corners, only receiving reinforcement for alternating )] TJ ET BT 26.250 422.169 Td /F1 9.8 Tf [(visits. For example, if corners 1 and 2 were active, an initial visit to corner 1 would be a correct visit and rewarded. To obtain )] TJ ET BT 26.250 410.264 Td /F1 9.8 Tf [(further reinforcement, the mouse was then required to visit corner 2. Repeat visits to corner 1 were classified as incorrect, and )] TJ ET BT 26.250 398.359 Td /F1 9.8 Tf [(mice would not receive a reward there. Following a visit to corner 2, the corners would switch again, such that reinforcement )] TJ ET BT 26.250 386.454 Td /F1 9.8 Tf [(would now be available only in corner 1 and so on. If a mouse incorrectly visits the same active corner more than once or visits )] TJ ET BT 26.250 374.550 Td /F1 9.8 Tf [(an exploratory corner after leaving an active corner, the identity of the target correct corner does not change and a visit to that )] TJ ET BT 26.250 362.645 Td /F1 9.8 Tf [(corner will result in available reinforcement; the only event leading to a switch in the correct corner identity is a visit to the )] TJ ET BT 26.250 350.740 Td /F1 9.8 Tf [(currently correct corner, in which reinforcement will be available.)] TJ ET BT 26.250 331.335 Td /F1 9.8 Tf [(Alternation data was calculated within a 113-s interval of leaving an active corner, such that only a visit to the correct corner 113-)] TJ ET BT 26.250 319.431 Td /F1 9.8 Tf [(s or less after exiting an active corner counted as a correct Alternation, with any visit to the currently )] TJ ET BT 458.682 319.431 Td /F5 9.8 Tf [(incorrect corner)] TJ ET BT 526.406 319.431 Td /F1 9.8 Tf [( counting )] TJ ET BT 26.250 307.526 Td /F1 9.8 Tf [(as incorrect, while visits to the )] TJ ET BT 158.460 307.526 Td /F5 9.8 Tf [(exploratory corners)] TJ ET BT 241.901 307.526 Td /F1 9.8 Tf [( are irrelevant. Inter visit intervals were cap to minimize the chances that other )] TJ ET BT 26.250 295.621 Td /F1 9.8 Tf [(events interrupted the alternating sequence the precise interval used is not of great significance, with similar results detected )] TJ ET BT 26.250 283.716 Td /F1 9.8 Tf [(with different intervals. Irrespective of this interval, visits to a correct corner were rewarded the interval is relevant only to the )] TJ ET BT 26.250 271.812 Td /F1 9.8 Tf [(data handling process. The primary measure employed to assess Alternation performance, percent alternation, is generated by )] TJ ET BT 26.250 259.907 Td /F1 9.8 Tf [(calculating the percentage of visits to active corners made following a visit to either of the two active corners within a particular )] TJ ET BT 26.250 248.002 Td /F1 9.8 Tf [(interval that are correct alternation visits. For example, if corners 1 and 2 are active, the number of visits to corner 2 following a )] TJ ET BT 26.250 236.097 Td /F1 9.8 Tf [(visit to corner 1 plus the number of visits to corner 1 following a visit to corner 2 comprises the total Alternation visits, which are )] TJ ET BT 26.250 224.193 Td /F1 9.8 Tf [(then divided by the total visits to either corner 1 or corner 2. Previous experiments in our laboratories have indicated that, )] TJ ET BT 26.250 212.288 Td /F1 9.8 Tf [(although mice do have some tendency to spontaneously alternate between foraging locations, their alternating behavior within )] TJ ET BT 26.250 200.383 Td /F1 9.8 Tf [(the corners is significantly increased by the type of enforced alternation training employed here, such that performance on this )] TJ ET BT 26.250 188.478 Td /F1 9.8 Tf [(task does appear to be sensitive to the animals learning the reinforcement contingencies.)] TJ ET BT 26.250 169.074 Td /F1 9.8 Tf [(Each corner contained two nosepoke recesses, one on each side, used to deliver water reinforcement during correct visits. )] TJ ET BT 26.250 157.169 Td /F1 9.8 Tf [(Each active corner delivered water only on one side, with the rewarded side dependant on the specific identity of the current )] TJ ET BT 26.250 145.264 Td /F1 9.8 Tf [(corner, such that if only the left side was rewarded in active corner 1, then only the right side was rewarded in active corner 2 )] TJ ET BT 26.250 133.359 Td /F1 9.8 Tf [(and vice versa. When mice made visits to a correct corner \(i.e., an alternation visit\) and nose-poked on the correct side, the )] TJ ET BT 26.250 121.455 Td /F1 9.8 Tf [(door behind the correct nosepoke recess for that corner opened for 8 s, providing access to water. After 8 s, the door closed, )] TJ ET BT 26.250 109.550 Td /F1 9.8 Tf [(preventing further access to water, with the mice then required to alternate and enter the other active corner for further rewards. )] TJ ET BT 26.250 97.645 Td /F1 9.8 Tf [(No penalty was imposed for initially nosepoking on the incorrect side; the first nosepoke during a correct visit to the correct side )] TJ ET BT 26.250 85.740 Td /F1 9.8 Tf [(always resulted in delivery of reinforcement. However, for the purposes of behavioral analysis, the proportion of initial )] TJ ET BT 26.250 73.836 Td /F1 9.8 Tf [(nosepokes to the correct side was calculated such that, for example, a mouse making a correct visit and subsequently )] TJ ET BT 26.250 61.931 Td /F1 9.8 Tf [(nosepoking to the wrong side and then to the correct side would receive water, but the mouses initial nosepoke for that visit )] TJ ET BT 26.250 50.026 Td /F1 9.8 Tf [(would be scored as incorrect \(from a behavioral efficiency perspective\).)] TJ ET Q q 225.000 0 0 82.500 35.250 684.750 cm /I3 Do Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(3)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Huntington Disease)] TJ ET Q endstream endobj 300 0 obj << /Type /Font /Subtype /Type1 /Name /F5 /BaseFont /Helvetica-Oblique /Encoding /WinAnsiEncoding >> endobj 301 0 obj << /Type /Annot /Subtype /Link /A 302 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 684.7500 260.2500 767.2500 ] >> endobj 302 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Figure1.jpg) >> endobj 303 0 obj << /Type /XObject /Subtype /Image /Width 300 /Height 110 /ColorSpace /DeviceRGB /Filter /DCTDecode /BitsPerComponent 8 /Length 11012>> stream JFIF;CREATOR: gd-jpeg v1.0 (using IJG JPEG v62), quality = 90 C     C   n," }!1AQa"q2#BR$3br %&'()*456789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz w!1AQaq"2B #3Rbr $4%&'()*56789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz ?i[~;Lת2 ʭ_oޟ,~%c+= ynY]hFỸ$?j n|?b\@ p8Oz]oG8$]s+ڙc2k?oqڸ˫] '1!2 Anڌ+i7p@sn ;?㟎Qķ`%p:}*]e5K)e\>B;~隃Ua< %iqi3ί[++(Y~s~W#w8U-^ye3O6*{c˟Ƥ8_#H<_/7u\dTa7;|Zִ{[zxY2#o#ּg}AGExž$&Ӭold4R]FwecϡY0¹W:^+!nvd2}Ԑ~1Ҽ.:=}ė +qϸzǥxJ3/KU>|xJSoھ]_] NӬwך;n8Tu'$ 7%"y0 8ϧOU>hd9T8Uwko\Ojٯ"Ǭ>#BZÝ*D\p?3MN~ \CH0y_z~O\-O?Oª(-\LҁߵQ` ZI.Əj+5R42HWnq|֟LExV/,Qra*f?~wx NfTSھ{BSX]Ǒn?kJmS4!ĞlߛI;wڻ+keH1>aP!"B f_ۺ  sԥàOaG8mDZrj)bRwdg#T4UXABnI RJ<ϷTFVU>_%@&@^0**:z拌uM-JXg YH3EYf"o* a ?(*dYcQ3UiWVyة[i0,ƹ7P4HWEb 짘%E0:@0} v VpFGEt X1?ݫſj(IT=xe']$ƯϛxV_  .gĉW&^V6ǯxrm';\ÑxM4ݝ_˸=b;b-W BFHi 2ҹ 5:\bKqx94 mV>kš`~5^]%c烵s/N ]A@l26A{kv˚؈ĹA$<׾|Gơvtɮ[3vu+zšm5m60_[G#ֺ! -M:Ӡ]+.kYT^2xBl ?_&|C\i\rH} Lzz㨯Y|9^_^j}ghTc޼x/ xtt5cNܱ+x 5cˏh)i o, k~'nIJ&[Y 4.X;d^kIx,ooZ<3)}~?)RŧGJ([{}+Ε-t;#{5=7kwWκ1iPI$ *A/kCŸ7xs+^boBsMCZ6uΡ^D6®@qQpx–xen0@vU&3'ڽA;TpOOP6ZivֵUB8OwwoKCPx>(${xRqtR9䍚D ݒbh"B?Cr1CA LT"GEiA@x/I֥LQFOb;zIWW;R5o5{/VHUYJkwP}V;>JJ9noAxG 0A!ֺO|M?\0?͟͟Je#&o|WkcZH|;WR~,UB2KqQ-䬏,~7x|v\o;E` ZKی)nߕpvt\}rخuac<:6LT=Sֺռ56iHp^}X5ZĞWMHFT)<5{b|5{N) r|eVφ"nh5[Qko,trL;@:huGs8h U$^1#y}{lC\VjI= _Ph G4@Tq_$~Կh_ޤr?j̻HrF pjfwZ?ķu^t4cm*G n~Mڴz<,#Ӡ?9.c83R5JRh[sA\@?hindg{>674[߳iKFQv}h{ hɯ}$,漉\1+^Zx;it?hڜQy2z|g?|`⇉4[ hitY1܇ҽEMGGF Dӥy4ɦro-HFssֺ].Ii-# 1,J5? ]iz9cn#=N}==$DIGz?i~Y^@Y#G_/p2F|7Ѽ| ,IބfT44*0[;2g dv?xUu\XvwcYb'`><;m|I]hhlY8VIȮ~ \(9B~0N @~Vh$imT`R*J+`XO=i XVDR0=\`91׮Έ.n$I8m^gLgj xkyѵ c/Bm!̷ݜ0;q}+,V&?.mOrnڔ#дJ IJ`v8~5c&3]Mp$d`22Qڰ9s'>M<(2!?>kоǢxoP4β}8bQvnd:x_ǟ"# ,I;ɩ<{W]ڙ4+s`ηbDw)\g_3|)nA㿴Hة*y_־QbfRcJБߎoMajF w"h?[űaԚDBzy=koI𥞃jb(N:^7x Niyocn~"kw-uR/ x[b}On#>ԓ2]+ſ-J6f;J%f;T /?gV]wQLc׸|_;[,c$뛲dvSy*Md\SzQkw{g|%ݣZ>-n$1[J#B`Y/7_ޱ{w3{2zWY[5*שO,]G3_Iߋ+ya)-2"R@#o4YA/N{Ep/Ο JsWUmڕΛqgWFw&,~q(SsRIEF6+Tl>;ӛ*qzwus5)cihu/ nXIMDb5f;&dc3I cԊAbd5nWG~x*䎥MjK[ JGH$J݌fB g$h/2F,`cu:~aPk-6HҼpK-ŵ⋷bIl~u eVɾEbQmcN6n PQY֝y?Sbd+ 3$wܸ#5iۈ>hL ɯz>$M;LOE3r@c~_']? ^bvŝ@` ~M]>x5-GbB(W3^O-LL1um1z9[UD# ++ƢвZOlrq%JnbÙ0v lnVxڴ@: \ܭf%ۏeyĎ#0SQQC|")$?uO ۟ڭoCzq T:`7N)Z; ;SJ p8?;Cg ٹ(B$Ԟ)%W#@ۭSrS$*21=Xn\ܥ|8!uUC,Ȳ/ ޹Y>ly'֮xĖ=WDr <+r>ip%f>V%N9:/sW Ac$UƔSqnZ20eI}Cs+G@[Vrq2+<3J[9)B9T>X2{UW&4,:~#Nu4sK",I \Jc HOTgx'kռ yd >G|~vo,6c̆^mطLaoK]`p ӥzVzΛ}i9֗M僑\z~TzT]-Yi Gd`ƩjN;TGcᩬB9%P9yЮ?-s{6e/G$Մr@Vv;3?|*vx~(P~Rz£o KO\Ҳ=jT5bKTtS2.䯒i(*{Dž)+_ċ}=: Rcj!s$rl5*QPk|,Jֶ:E)09|/uH<™N Ƹ MjQɥjMj.$򮬭̈SqA~$],:f;'$f\+1%%kjʤdm&./ʕW)3KoxI_QOj5ut;!s$b{ʴ3Iu&hy'/̸c qHۙ1{]g8Z֗""9`xLf=~r>#8e07jp\#8l>Ű6X.)6ҕ=8'S q1K0:՟=ŽރLVGҽb#'~^gmؘd9n/mԠğ+^-&}BUiNp6p;:VjcBA[TKe<@mwxIKA ȓG$wy 'Úwp\CEo lB?1%5_[8taýj#oCvU/=f2W8u5VQVc }A?X+,m~) $)$G}:WX_ y,qx`{RI S}ht(1ZHHH,)zwEE\N#k0h>扩5xŦ.R.2e[D13Alc[*Z3xx9][^*o"Y"G0Lnr?{y?0dY[N^_SogwP:rz f;9|5ZKc# &I  iU:*;m f.HێDJ #9_+ rQYd,r,A\s^ҢN]%]g|OTG}EuB( ۡ3V[)c#K,}S ҁor_5Ԗ^t{UVkf}%fB2DZr\$:9R q^)iϦ]C͗*M˒@>g{jY?isi mslcl;F=S_I1|f.GahEFbM[&fjjzI"vi\~NqsWz\Z,cC9_[JnMwDA{)] 8Sӌrg3" ދ?ܤ0#d*$*[7c ڍ܍J͸+ ?fr6r_g-_^\G3Fwќ 'Өٟ$%&f2nY~i&լg>_)<3wXtSX+cM?>z>?o^?7lk{[hJBPs^ciPhW~+o%5i01j|;tJ%/'޾ݺ4 FH#x}o%G@\D5uR?.^b'"<3ԟw6#Nzzؐu/O/ |SW" iy=yY\]:r3e}_Li?hBA-Qp" ֏="*@q(h|-K>}8H8RodxSQdS(i9K,s!khĭb=Q^pX&39?w4-"R֘eE)gNoMUŤ8G,ֹ+&9<z}+2Ȳ&it /v5o$֫9 3ʀI)& N5qJ̚霯¶V[=ET`+!GJM>g*HO |S&Cg,>87cZEӏC)cvs/$TE m>s(SCKKܺ#.@8**?#^ѫ|4U\Ku]Jw<,O=p{΁ӞFإpK \ʥ 4O. ISΌǧ7s6}>HwO4ZM$Ŕ)#׶ x~kh-ه\<ӎ=a vD' wA.~,7A c~z{U.eɸcþ/3 q^}1ӭ~14Q" dیߥyݟv,s$}k_كxr}y|H|J ]6lǕqNQ-^Z~%k endstream endobj 304 0 obj << /Type /Annot /Subtype /Link /A 305 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 684.7500 260.2500 767.2500 ] >> endobj 305 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Figure1.jpg) >> endobj 306 0 obj << /Type /Annot /Subtype /Link /A 307 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 684.7500 260.2500 767.2500 ] >> endobj 307 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Figure1.jpg) >> endobj 308 0 obj << /Type /Page /Parent 3 0 R /Annots [ 310 0 R 313 0 R 316 0 R 318 0 R 320 0 R 322 0 R ] /Contents 309 0 R >> endobj 309 0 obj << /Length 14233 >> stream 0.271 0.267 0.267 rg q 15.000 40.204 577.500 736.796 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F5 9.8 Tf [(Reversal:)] TJ ET BT 67.424 767.476 Td /F1 9.8 Tf [( The final phase is a nosepoke reversal \(Figure 2 right panel\). The protocol is similar to the Alternation phase, except )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(that the correct nosepoke recesses are switched between the two active corners, such that if the left nosepoke was correct in )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(corner 1, instead the right nosepoke was correct no differences between the genotypes were observed in performance on this )] TJ ET BT 26.250 731.762 Td /F1 9.8 Tf [(task so these data are not presented, but this information is included here for completeness.)] TJ ET 0.965 0.965 0.965 rg 26.250 472.835 555.000 249.046 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 721.881 m 581.250 721.881 l 581.250 721.131 l 26.250 721.131 l f 26.250 472.835 m 581.250 472.835 l 581.250 473.585 l 26.250 473.585 l f q 450.000 0 0 171.000 35.250 541.131 cm /I4 Do Q q 35.250 484.085 537.000 51.046 re W n 0.271 0.267 0.267 rg BT 35.250 525.607 Td /F4 9.8 Tf [(Fig. 2: Graphical depiction of the IntelliCage and the experimental procedures: Habituation, Alternation, and )] TJ ET BT 35.250 513.702 Td /F4 9.8 Tf [(Reversal.)] TJ ET BT 35.250 494.332 Td /F1 9.8 Tf [(Shaded circles signal the armed receptacles. NP: Nosepoke)] TJ ET Q BT 26.250 455.811 Td /F4 9.8 Tf [(Experiment 1. )] TJ ET BT 92.901 455.811 Td /F1 9.8 Tf [(An initial experiment was carried out in order to evaluate both the overall phenotype of the R6/2 mouse and the )] TJ ET BT 26.250 443.907 Td /F1 9.8 Tf [(specific capacity of both wild-type \(WT\) and R6/2 mice to learn a simple alternation rule, requiring them to switch between two )] TJ ET BT 26.250 432.002 Td /F1 9.8 Tf [(locations to retrieve water reinforcement.)] TJ ET BT 202.355 432.002 Td /F1 9.8 Tf [(R6/2 mice were tested weekly from 7 to 10 weeks old. The subjects were a cohort of )] TJ ET BT 26.250 420.097 Td /F1 9.8 Tf [(44 nave female mice evenly split by genotype in four groups of 11 mice each. Each group was tested in one cage. )] TJ ET BT 26.250 400.692 Td /F1 9.8 Tf [(Animals were water deprived in their home cages for 16 h prior to each phase of testing in the PhenoCube)] TJ ET BT 484.178 404.581 Td /F1 8.7 Tf [(TM)] TJ ET BT 496.693 400.692 Td /F1 9.8 Tf [( system. During )] TJ ET BT 26.250 388.788 Td /F1 9.8 Tf [(this period and the rest of the experiment feed were available ad libitum. The initial 7-wk test was split into three sections \(as )] TJ ET BT 26.250 376.883 Td /F1 9.8 Tf [(illustrated in Figure 3\), with an initial 6-h Habituation day to acclimate the mice to the cage, followed by a 30-h session on the )] TJ ET BT 26.250 364.978 Td /F1 9.8 Tf [(Alternation protocol and then a separate 6-h session with the Reversal protocol. The remaining test weeks were split into two 30-)] TJ ET BT 26.250 353.073 Td /F1 9.8 Tf [(h sessions \(Figure 3\), with the nosepoke task reversed at the start of each session relative to the previous session. Despite the )] TJ ET BT 26.250 341.169 Td /F1 9.8 Tf [(regular test schedule, some R6/2 mice still appeared unable to maintain licking behavior on the Alternation protocol; non-licking )] TJ ET BT 26.250 329.264 Td /F1 9.8 Tf [(animals were switched to the Habituation protocol and ultimately removed from the cage as necessary. The switch to )] TJ ET BT 26.250 317.359 Td /F1 9.8 Tf [(Habituation protocol was implemented electronically based on the RFID chip without disturbing the group or changing the )] TJ ET BT 26.250 305.454 Td /F1 9.8 Tf [(protocol for the other mice in the social environment. The number of mutant mice still licking on the Alternation procedure at the )] TJ ET BT 26.250 293.550 Td /F1 9.8 Tf [(end of each weeks testing \(at 7, 8, 9 and 10 weeks\) were 20, 12, 6, and 4 mice respectively. Note that when mice were )] TJ ET BT 26.250 281.645 Td /F1 9.8 Tf [(switched to the Habitation protocol, they frequently resumed licking, indicating that their failure to lick on Alternation does not )] TJ ET BT 26.250 269.740 Td /F1 9.8 Tf [(simply reflect a motor deficit.)] TJ ET 0.965 0.965 0.965 rg 26.250 47.704 555.000 212.155 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 259.859 m 581.250 259.859 l 581.250 259.109 l 26.250 259.109 l f 26.250 47.704 m 581.250 47.704 l 581.250 48.454 l 26.250 48.454 l f q 450.000 0 0 167.250 35.250 82.859 cm /I5 Do Q q 35.250 58.954 537.000 17.905 re W n 0.271 0.267 0.267 rg BT 35.250 67.335 Td /F4 9.8 Tf [(Fig. 3: R6/2 mice test procedure.)] TJ ET Q Q q 15.000 40.204 577.500 736.796 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F5 9.8 Tf [(Reversal:)] TJ ET BT 67.424 767.476 Td /F1 9.8 Tf [( The final phase is a nosepoke reversal \(Figure 2 right panel\). The protocol is similar to the Alternation phase, except )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(that the correct nosepoke recesses are switched between the two active corners, such that if the left nosepoke was correct in )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(corner 1, instead the right nosepoke was correct no differences between the genotypes were observed in performance on this )] TJ ET BT 26.250 731.762 Td /F1 9.8 Tf [(task so these data are not presented, but this information is included here for completeness.)] TJ ET 0.965 0.965 0.965 rg 26.250 472.835 555.000 249.046 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 721.881 m 581.250 721.881 l 581.250 721.131 l 26.250 721.131 l f 26.250 472.835 m 581.250 472.835 l 581.250 473.585 l 26.250 473.585 l f q 450.000 0 0 171.000 35.250 541.131 cm /I4 Do Q q 35.250 484.085 537.000 51.046 re W n 0.271 0.267 0.267 rg BT 35.250 525.607 Td /F4 9.8 Tf [(Fig. 2: Graphical depiction of the IntelliCage and the experimental procedures: Habituation, Alternation, and )] TJ ET BT 35.250 513.702 Td /F4 9.8 Tf [(Reversal.)] TJ ET BT 35.250 494.332 Td /F1 9.8 Tf [(Shaded circles signal the armed receptacles. NP: Nosepoke)] TJ ET Q BT 26.250 455.811 Td /F4 9.8 Tf [(Experiment 1. )] TJ ET BT 92.901 455.811 Td /F1 9.8 Tf [(An initial experiment was carried out in order to evaluate both the overall phenotype of the R6/2 mouse and the )] TJ ET BT 26.250 443.907 Td /F1 9.8 Tf [(specific capacity of both wild-type \(WT\) and R6/2 mice to learn a simple alternation rule, requiring them to switch between two )] TJ ET BT 26.250 432.002 Td /F1 9.8 Tf [(locations to retrieve water reinforcement.)] TJ ET BT 202.355 432.002 Td /F1 9.8 Tf [(R6/2 mice were tested weekly from 7 to 10 weeks old. The subjects were a cohort of )] TJ ET BT 26.250 420.097 Td /F1 9.8 Tf [(44 nave female mice evenly split by genotype in four groups of 11 mice each. Each group was tested in one cage. )] TJ ET BT 26.250 400.692 Td /F1 9.8 Tf [(Animals were water deprived in their home cages for 16 h prior to each phase of testing in the PhenoCube)] TJ ET BT 484.178 404.581 Td /F1 8.7 Tf [(TM)] TJ ET BT 496.693 400.692 Td /F1 9.8 Tf [( system. During )] TJ ET BT 26.250 388.788 Td /F1 9.8 Tf [(this period and the rest of the experiment feed were available ad libitum. The initial 7-wk test was split into three sections \(as )] TJ ET BT 26.250 376.883 Td /F1 9.8 Tf [(illustrated in Figure 3\), with an initial 6-h Habituation day to acclimate the mice to the cage, followed by a 30-h session on the )] TJ ET BT 26.250 364.978 Td /F1 9.8 Tf [(Alternation protocol and then a separate 6-h session with the Reversal protocol. The remaining test weeks were split into two 30-)] TJ ET BT 26.250 353.073 Td /F1 9.8 Tf [(h sessions \(Figure 3\), with the nosepoke task reversed at the start of each session relative to the previous session. Despite the )] TJ ET BT 26.250 341.169 Td /F1 9.8 Tf [(regular test schedule, some R6/2 mice still appeared unable to maintain licking behavior on the Alternation protocol; non-licking )] TJ ET BT 26.250 329.264 Td /F1 9.8 Tf [(animals were switched to the Habituation protocol and ultimately removed from the cage as necessary. The switch to )] TJ ET BT 26.250 317.359 Td /F1 9.8 Tf [(Habituation protocol was implemented electronically based on the RFID chip without disturbing the group or changing the )] TJ ET BT 26.250 305.454 Td /F1 9.8 Tf [(protocol for the other mice in the social environment. The number of mutant mice still licking on the Alternation procedure at the )] TJ ET BT 26.250 293.550 Td /F1 9.8 Tf [(end of each weeks testing \(at 7, 8, 9 and 10 weeks\) were 20, 12, 6, and 4 mice respectively. Note that when mice were )] TJ ET BT 26.250 281.645 Td /F1 9.8 Tf [(switched to the Habitation protocol, they frequently resumed licking, indicating that their failure to lick on Alternation does not )] TJ ET BT 26.250 269.740 Td /F1 9.8 Tf [(simply reflect a motor deficit.)] TJ ET 0.965 0.965 0.965 rg 26.250 47.704 555.000 212.155 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 259.859 m 581.250 259.859 l 581.250 259.109 l 26.250 259.109 l f 26.250 47.704 m 581.250 47.704 l 581.250 48.454 l 26.250 48.454 l f q 450.000 0 0 167.250 35.250 82.859 cm /I5 Do Q q 35.250 58.954 537.000 17.905 re W n 0.271 0.267 0.267 rg BT 35.250 67.335 Td /F4 9.8 Tf [(Fig. 3: R6/2 mice test procedure.)] TJ ET Q Q q 15.000 40.204 577.500 736.796 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F5 9.8 Tf [(Reversal:)] TJ ET BT 67.424 767.476 Td /F1 9.8 Tf [( The final phase is a nosepoke reversal \(Figure 2 right panel\). The protocol is similar to the Alternation phase, except )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(that the correct nosepoke recesses are switched between the two active corners, such that if the left nosepoke was correct in )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(corner 1, instead the right nosepoke was correct no differences between the genotypes were observed in performance on this )] TJ ET BT 26.250 731.762 Td /F1 9.8 Tf [(task so these data are not presented, but this information is included here for completeness.)] TJ ET 0.965 0.965 0.965 rg 26.250 472.835 555.000 249.046 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 721.881 m 581.250 721.881 l 581.250 721.131 l 26.250 721.131 l f 26.250 472.835 m 581.250 472.835 l 581.250 473.585 l 26.250 473.585 l f q 450.000 0 0 171.000 35.250 541.131 cm /I4 Do Q q 35.250 484.085 537.000 51.046 re W n 0.271 0.267 0.267 rg BT 35.250 525.607 Td /F4 9.8 Tf [(Fig. 2: Graphical depiction of the IntelliCage and the experimental procedures: Habituation, Alternation, and )] TJ ET BT 35.250 513.702 Td /F4 9.8 Tf [(Reversal.)] TJ ET BT 35.250 494.332 Td /F1 9.8 Tf [(Shaded circles signal the armed receptacles. NP: Nosepoke)] TJ ET Q BT 26.250 455.811 Td /F4 9.8 Tf [(Experiment 1. )] TJ ET BT 92.901 455.811 Td /F1 9.8 Tf [(An initial experiment was carried out in order to evaluate both the overall phenotype of the R6/2 mouse and the )] TJ ET BT 26.250 443.907 Td /F1 9.8 Tf [(specific capacity of both wild-type \(WT\) and R6/2 mice to learn a simple alternation rule, requiring them to switch between two )] TJ ET BT 26.250 432.002 Td /F1 9.8 Tf [(locations to retrieve water reinforcement.)] TJ ET BT 202.355 432.002 Td /F1 9.8 Tf [(R6/2 mice were tested weekly from 7 to 10 weeks old. The subjects were a cohort of )] TJ ET BT 26.250 420.097 Td /F1 9.8 Tf [(44 nave female mice evenly split by genotype in four groups of 11 mice each. Each group was tested in one cage. )] TJ ET BT 26.250 400.692 Td /F1 9.8 Tf [(Animals were water deprived in their home cages for 16 h prior to each phase of testing in the PhenoCube)] TJ ET BT 484.178 404.581 Td /F1 8.7 Tf [(TM)] TJ ET BT 496.693 400.692 Td /F1 9.8 Tf [( system. During )] TJ ET BT 26.250 388.788 Td /F1 9.8 Tf [(this period and the rest of the experiment feed were available ad libitum. The initial 7-wk test was split into three sections \(as )] TJ ET BT 26.250 376.883 Td /F1 9.8 Tf [(illustrated in Figure 3\), with an initial 6-h Habituation day to acclimate the mice to the cage, followed by a 30-h session on the )] TJ ET BT 26.250 364.978 Td /F1 9.8 Tf [(Alternation protocol and then a separate 6-h session with the Reversal protocol. The remaining test weeks were split into two 30-)] TJ ET BT 26.250 353.073 Td /F1 9.8 Tf [(h sessions \(Figure 3\), with the nosepoke task reversed at the start of each session relative to the previous session. Despite the )] TJ ET BT 26.250 341.169 Td /F1 9.8 Tf [(regular test schedule, some R6/2 mice still appeared unable to maintain licking behavior on the Alternation protocol; non-licking )] TJ ET BT 26.250 329.264 Td /F1 9.8 Tf [(animals were switched to the Habituation protocol and ultimately removed from the cage as necessary. The switch to )] TJ ET BT 26.250 317.359 Td /F1 9.8 Tf [(Habituation protocol was implemented electronically based on the RFID chip without disturbing the group or changing the )] TJ ET BT 26.250 305.454 Td /F1 9.8 Tf [(protocol for the other mice in the social environment. The number of mutant mice still licking on the Alternation procedure at the )] TJ ET BT 26.250 293.550 Td /F1 9.8 Tf [(end of each weeks testing \(at 7, 8, 9 and 10 weeks\) were 20, 12, 6, and 4 mice respectively. Note that when mice were )] TJ ET BT 26.250 281.645 Td /F1 9.8 Tf [(switched to the Habitation protocol, they frequently resumed licking, indicating that their failure to lick on Alternation does not )] TJ ET BT 26.250 269.740 Td /F1 9.8 Tf [(simply reflect a motor deficit.)] TJ ET 0.965 0.965 0.965 rg 26.250 47.704 555.000 212.155 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 259.859 m 581.250 259.859 l 581.250 259.109 l 26.250 259.109 l f 26.250 47.704 m 581.250 47.704 l 581.250 48.454 l 26.250 48.454 l f q 450.000 0 0 167.250 35.250 82.859 cm /I5 Do Q q 35.250 58.954 537.000 17.905 re W n 0.271 0.267 0.267 rg BT 35.250 67.335 Td /F4 9.8 Tf [(Fig. 3: R6/2 mice test procedure.)] TJ ET Q Q q 450.000 0 0 171.000 35.250 541.131 cm /I4 Do Q q 450.000 0 0 167.250 35.250 82.859 cm /I5 Do Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(4)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Huntington Disease)] TJ ET Q endstream endobj 310 0 obj << /Type /Annot /Subtype /Link /A 311 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 541.1310 485.2500 712.1310 ] >> endobj 311 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Balci_fig2.jpg) >> endobj 312 0 obj << /Type /XObject /Subtype /Image /Width 600 /Height 228 /ColorSpace /DeviceRGB /Filter /DCTDecode /BitsPerComponent 8 /Length 27226>> stream JFIFddC  !"$"$CX"W !1A"Qa2T#78BRSUqvt$3rFbu%&4CVs5D)!1AQa"# ?.$.&@ܡfCli ]xCJGj 7K͇6 ܦmGijl\<)s/Pt㞄t^Nmr7rǚ̢[!낃!6![dloV$6V7Lle[.*zbfJ+tӡh%)_<{PnB&_HAan)R~ + Ųy uJO:A׸ק Q;3ugqUӆ(:8ׅ#wNMce1O&C:RZOM$O gyboER4%7\|;}nZvޙ2m Q;3t~kzW˻>t|?p-$: H-z61`[aWZmn,믨c]H]v1ޓKMg٦V-Y.?8TqdJ|I@ WXԋzlJkC WqZٿח"spm}JBN<?yُ&Ef#l1ڄ\KТ6I J9^}nj7qy>?rKiB[qL%!kiF^=D $aZٗG]KҢ-- U!ED+iӠ8]ś.Z]@vi2?% (!*d=Я|(O'P|1LhSb&Ϙ [)Jij%%D]]8iR&:žd%l!HiҰsvMoߔO0W8=lL!ٕsÒv1RjڜQI#'+c% $Y(:qDkQ h <(\wK,mIuah=5҂j6ڬdeXKZw[#uӆ(:&\ɗR^>=eZ$( {;AŸAft Ȍum+IAPT:pߚó?O7Gw3o$Ȅۯn;ǝX(I=O}܆v-CrVԗjW~Uuݏ[[uӆ(:|t5?fN9.7eHuvZp);kƧMQ;B\Fr뼛MwNd8sѕE!CHH|$Ֆ]q>韪1]~I߫,0ӔqG23mב"Hm+&?~:pߚó?L{՞"7G׃,6ӫ%W5%.H uŎi m^͹B tן{XݕU~:pߚó?O7GwgqmvZʤ(蔐{nq 2A!mh _pCm Ύn1]hZ'r0ۖ9HSؐ r>5p!eM&yK(HI :tOV=Dx~Q?VulIg˔ KЧ>Ɛ$']u> 7g-9dbJ|RJ9I(NQV=Dx~Q?Ď E_fV$8S>\h\%|R /~N-lmML3Nz\7׫tAov%}]z=!ӆ(:8^,/Yqa=TN}u=ާ|͞Qr;#L=:Һ-Gmc@㡠~k>:pߚó?VʸMZKh u=BI=wOC<b! :?5*cU*8orٟӧN\vg[.̆P2b(%$i+T}DƓ^Ŭ>[-c!8! =Yf}^Wpd֧P%%RM"DO26Fޣ:pߚó?^Ooj]uxt5?fN6a@ɷLYnrgC6erַt4^b=Z'ʽ-s҅3!~:^oYޡU~k>:pߚó?_+#2lnH\0dZ%+J碴}e3˄b+jdKiq埒#kQ l ui1;VV\qr2ۖ9HS$)f_d7!}#ƪX n&gS')<m{P5q/^]kɜ] -LⰏ=GT q3ľ1vM4\4#\m.$%#jV=Dx~Q? gj}jq{ JSt+Rdhr^"q7ێΦژǦq[uHwu~Q?ߔO.Mc5ťi r㣝Ҋ' (R}e{Mq@8 %⡩K\$@Ct<(O'Ped7!}#Ɵd7!}#Ƶ/'W|%xn7ϯWc3;?ܾs.GW#d4|PuԇN\vg:n-yrg8<aʏU'WS؝=9l'#WRj\ǒzt~k>:pߚó?RYFlO RJKhNoRT6s;psZQVh-ґkVL 94n|t5?fN8orٟӮhɡJC&=$(Q)>aI UZYf-c| Wwb?Q2":](AZSa)'^)ξ|,sEժRR˸9M?[?V]^[^)7,W<w8{:T#Aw֥+SL>zɯIn),#r]%:OS]W|g9H.W2myJ+JMƍg]S<'z'ؑV Oe)V8f9)_Irs=9JU)JP)JP)JP)JPW2kDw#I"&\ H(Cağ:12P͚tfpOX$ڟQ# l;k=ceLAnK'DZOpG.⥖LT.P$olRv׻FyW*VBqDJ.M3<':BTyuRQ=4nt r GE2OPJWS"y`Ûq>\&KaQj\GjqE\7$P̅GL8ZKG:X=y=cqc7!P ]*>)iOjm޳[k9&?yzD|tɮؿ|$Ֆ]q[>韪1]~I߫,0Ǿy;O*T8!KVBS(HIVZ2H/ʽrPfP=K? iΕ:#=9ni݌-ERBjr c! 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xKҾ*xb3s?5o<"A IJC"';HH?>8=-6So[{k;t 2xWĿFγ=杯xB > "u c^7 ?eמZUֹ<~Tޯ(TU@{QJCu#v7/i90$v@|3^`b.`}&@Of8 k 2 ?29x ޾N[iv^OCs3Dsz9 dx&/=vSzbN;k?nǂ|GZ|D mEV(;i+;n ᯆ>+bþ.m5~=˞̧A|O~ jzv>"k#0]ClX㧁In ޡ0;:x_O< ^h\6Mk+ 5v n4ʈֵID/C`*/EP{ր+X&p>2qwBgsOmbCsg7(^jLȳG19i +蚡emP[Y[6;l%l zkB񘗊k%"*GKI\EQ@Z;QI@ EPEPE(Q@袀 ( ;QF(((ih,=JV6vdoYj"[晘1=^<?5kdmP1 < @qU簂Xe/PSǭn4y2\cx]B_qpJyҴ7t=;Ps%QLKnLWF(ܛ\ ;Et4 b (1ڄs3^9w[¾ K]KHg#C 1#72 r8^F+HMc9Osl-|Ug}%ԓ^Jmd77[~g- !#w%&2?=0qEw>'ҥmO9o 2Ļ6f$IK%p*~Qɧm$B)-VFEPE +>v١pFI$*l&y4=jH+qiiqGu4LwB#!l;`yc_Ks=܎Mg%YH\@ǜ֫Ɍ{Vꮖ93hw:3i;¨y g>vqSn#e6ŠAEPE9S~-qus\A IݣeQ\w4mbn^CkoaKbc% JG)=)$z("625#'9'K'!{$No(XrNC]_4VF:z5̆hUb2T ԭ t+ kr7ԁǹF̐$qڝ1F+#`(Q@s9 /<݂ ̌EE4%I\qidpXҬM41wmA3| /=i6Z^B +p$3ֽVn%!%GU'x٣/egt:?/|y{hll1đFmfG$c%1AJ:QY9s[1oQ@IaEP endstream endobj 316 0 obj << /Type /Annot /Subtype /Link /A 317 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 541.1310 485.2500 712.1310 ] >> endobj 317 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Balci_fig2.jpg) >> endobj 318 0 obj << /Type /Annot /Subtype /Link /A 319 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 82.8592 485.2500 250.1092 ] >> endobj 319 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Balci_fig3.jpg) >> endobj 320 0 obj << /Type /Annot /Subtype /Link /A 321 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 541.1310 485.2500 712.1310 ] >> endobj 321 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Balci_fig2.jpg) >> endobj 322 0 obj << /Type /Annot /Subtype /Link /A 323 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 82.8592 485.2500 250.1092 ] >> endobj 323 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Balci_fig3.jpg) >> endobj 324 0 obj << /Type /Page /Parent 3 0 R /Annots [ 326 0 R 329 0 R 331 0 R ] /Contents 325 0 R >> endobj 325 0 obj << /Length 19413 >> stream 0.271 0.267 0.267 rg q 15.000 26.932 577.500 750.068 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F4 9.8 Tf [(Experiment 2. )] TJ ET BT 92.901 767.476 Td /F1 9.8 Tf [(BACHD mice were evaluated longitudinally, with testing at 16, 24, 36, and 52 weeks of age. The subjects were a )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(cohort of 64 nave female mice, split evenly into four cages of 8 mice per genotype \(a total of eight cages\), each tested in a )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(single PhenoCube)] TJ ET BT 105.917 747.555 Td /F1 8.7 Tf [(TM)] TJ ET BT 118.432 743.667 Td /F1 9.8 Tf [(. In contrast to the R6/2 study, the animals were tested in a single long experimental session at each test )] TJ ET BT 26.250 731.762 Td /F1 9.8 Tf [(age, with the procedures employed otherwise as described for the R6/2 experiment. The timeline for these test sessions is )] TJ ET BT 26.250 719.857 Td /F1 9.8 Tf [(summarized in Figure 4, below. Note that, during the initial test at 16 weeks, the mice received an initial 24 hour period on the )] TJ ET BT 26.250 707.952 Td /F1 9.8 Tf [(simple Habituation protocol, to allow them to become accustomed to the cage environment, before switching to Alternation for )] TJ ET BT 26.250 696.048 Td /F1 9.8 Tf [(48 hours. In the later tests, the mice were placed immediately onto the Alternation protocol, with the corner contingency )] TJ ET BT 26.250 684.143 Td /F1 9.8 Tf [(reversing after 24 hours.)] TJ ET 0.965 0.965 0.965 rg 26.250 456.107 555.000 218.155 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 674.262 m 581.250 674.262 l 581.250 673.512 l 26.250 673.512 l f 26.250 456.107 m 581.250 456.107 l 581.250 456.857 l 26.250 456.857 l f q 450.000 0 0 173.250 35.250 491.262 cm /I6 Do Q q 35.250 467.357 537.000 17.905 re W n 0.271 0.267 0.267 rg BT 35.250 475.738 Td /F4 9.8 Tf [(Fig. 4: BACHD mice test procedure.)] TJ ET Q BT 26.250 439.083 Td /F4 9.8 Tf [(Computer vision:)] TJ ET BT 26.250 419.679 Td /F1 9.8 Tf [(In addition to data collected through the Intellicage hardware \(visits to each of the four corners\) general activity data was )] TJ ET BT 26.250 407.774 Td /F1 9.8 Tf [(collected through PsychoGenics Incs proprietary computer vision automated video scoring system. Our computer vision detects )] TJ ET BT 26.250 395.869 Td /F1 9.8 Tf [(and separates individual animals in the central area of the cage. As all mice in a cage had the same genotype we did not track )] TJ ET BT 26.250 383.964 Td /F1 9.8 Tf [(their identity across the whole session time, although the system was developed since to do so; accordingly, these data are )] TJ ET BT 26.250 372.060 Td /F1 9.8 Tf [(pooled across individuals within a cage. The primary measures currently implemented in this system are outlined below \(note )] TJ ET BT 26.250 360.155 Td /F1 9.8 Tf [(that animals within a corner are invisible to the camera and are accordingly not scored\):)] TJ ET BT 26.250 340.750 Td /F5 9.8 Tf [(Locomotion: )] TJ ET BT 82.069 340.750 Td /F1 9.8 Tf [(Measured as the distance travelled and normalized by the number of mice detected in each frame.)] TJ ET BT 26.250 321.345 Td /F5 9.8 Tf [(Immobility:)] TJ ET BT 72.835 321.345 Td /F1 9.8 Tf [( Measured as the time when an individual mouse was detected not moving \(maximum speed cutoff: 0.7 cm/s\).)] TJ ET BT 26.250 301.941 Td /F5 9.8 Tf [(Climbing:)] TJ ET BT 66.878 301.941 Td /F1 9.8 Tf [( Scored when a mouse was detected climbing on either of the climbing rods along the long wall of the cage or on the )] TJ ET BT 26.250 290.036 Td /F1 9.8 Tf [(climbing structure \(see Figure 1, left panel\).)] TJ ET BT 26.250 270.631 Td /F5 9.8 Tf [(Rearing:)] TJ ET BT 63.095 270.631 Td /F1 9.8 Tf [( Scored when an individual mouse was detected rearing against the walls of the apparatus.)] TJ ET BT 26.250 251.226 Td /F4 9.8 Tf [(Data analysis:)] TJ ET BT 26.250 231.822 Td /F1 9.8 Tf [(We analyzed data with mixed-design three-way ANOVA with SAS \(SAS Institute Inc.\) using Mixed Effect Models, based on )] TJ ET BT 26.250 219.917 Td /F1 9.8 Tf [(likelihood estimation. The models were fitted using the procedure PROC MIXED using genotype as the between-subjects )] TJ ET BT 26.250 208.012 Td /F1 9.8 Tf [(variable and light cycle and age as within-subjects variable. Significant interactions were followed by the simple effects analysis. )] TJ ET BT 26.250 196.107 Td /F1 9.8 Tf [(Where multiple interactions are present involving a given factor, simple effects analyses were conducted only for the highest )] TJ ET BT 26.250 184.203 Td /F1 9.8 Tf [(order interaction. An effect was considered significant if )] TJ ET BT 267.397 184.203 Td /F5 9.8 Tf [(p)] TJ ET BT 272.818 184.203 Td /F1 9.8 Tf [( < 0.05.)] TJ ET BT 26.250 147.600 Td /F4 12.0 Tf [(Results)] TJ ET BT 26.250 127.646 Td /F4 9.8 Tf [(Experiment 1. Analysis of R6/2 mice)] TJ ET BT 193.150 127.646 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 108.241 Td /F5 9.8 Tf [(Corner Entries: )] TJ ET BT 94.519 108.241 Td /F1 9.8 Tf [(R6/2 and WT mice did not exhibit differential frequency of corner entries. Both groups visited the corners equally )] TJ ET BT 26.250 96.336 Td /F1 9.8 Tf [(often, showed more visits during the dark light cycle, and reduced corner entries with repeated testing \(Figure 5A; Table I; Age )] TJ ET BT 26.250 84.432 Td /F1 9.8 Tf [(and Cycle main effects: )] TJ ET BT 130.292 84.432 Td /F5 9.8 Tf [(p)] TJ ET BT 135.713 84.432 Td /F1 9.8 Tf [(s < 0.05; simple effects analysis: )] TJ ET BT 279.048 84.432 Td /F5 9.8 Tf [(ps < )] TJ ET BT 300.459 84.432 Td /F1 9.8 Tf [(0)] TJ ET BT 305.880 84.432 Td /F5 9.8 Tf [(.)] TJ ET BT 308.591 84.432 Td /F1 9.8 Tf [(0001\). R6/2 mice showed slightly larger differences across the )] TJ ET BT 26.250 72.527 Td /F1 9.8 Tf [(day cycle although the differences between groups were not significant within each phase \(Cycle x Genotype interaction: )] TJ ET BT 548.665 72.527 Td /F5 9.8 Tf [(p )] TJ ET BT 556.796 72.527 Td /F1 9.8 Tf [(< )] TJ ET BT 26.250 60.622 Td /F1 9.8 Tf [(0.05; simple effects analysis: )] TJ ET BT 153.595 60.622 Td /F5 9.8 Tf [(p)] TJ ET BT 159.016 60.622 Td /F1 9.8 Tf [(s > 0.11\).)] TJ ET BT 26.250 41.217 Td /F5 9.8 Tf [(Licking:)] TJ ET BT 59.303 41.217 Td /F1 9.8 Tf [( Analysis of licking behavior pointed at increased thirst in R6/2 mice during the dark period. R6/2 mice licked more )] TJ ET Q q 15.000 26.932 577.500 750.068 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F4 9.8 Tf [(Experiment 2. )] TJ ET BT 92.901 767.476 Td /F1 9.8 Tf [(BACHD mice were evaluated longitudinally, with testing at 16, 24, 36, and 52 weeks of age. The subjects were a )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(cohort of 64 nave female mice, split evenly into four cages of 8 mice per genotype \(a total of eight cages\), each tested in a )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(single PhenoCube)] TJ ET BT 105.917 747.555 Td /F1 8.7 Tf [(TM)] TJ ET BT 118.432 743.667 Td /F1 9.8 Tf [(. In contrast to the R6/2 study, the animals were tested in a single long experimental session at each test )] TJ ET BT 26.250 731.762 Td /F1 9.8 Tf [(age, with the procedures employed otherwise as described for the R6/2 experiment. The timeline for these test sessions is )] TJ ET BT 26.250 719.857 Td /F1 9.8 Tf [(summarized in Figure 4, below. Note that, during the initial test at 16 weeks, the mice received an initial 24 hour period on the )] TJ ET BT 26.250 707.952 Td /F1 9.8 Tf [(simple Habituation protocol, to allow them to become accustomed to the cage environment, before switching to Alternation for )] TJ ET BT 26.250 696.048 Td /F1 9.8 Tf [(48 hours. In the later tests, the mice were placed immediately onto the Alternation protocol, with the corner contingency )] TJ ET BT 26.250 684.143 Td /F1 9.8 Tf [(reversing after 24 hours.)] TJ ET 0.965 0.965 0.965 rg 26.250 456.107 555.000 218.155 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 674.262 m 581.250 674.262 l 581.250 673.512 l 26.250 673.512 l f 26.250 456.107 m 581.250 456.107 l 581.250 456.857 l 26.250 456.857 l f q 450.000 0 0 173.250 35.250 491.262 cm /I6 Do Q q 35.250 467.357 537.000 17.905 re W n 0.271 0.267 0.267 rg BT 35.250 475.738 Td /F4 9.8 Tf [(Fig. 4: BACHD mice test procedure.)] TJ ET Q BT 26.250 439.083 Td /F4 9.8 Tf [(Computer vision:)] TJ ET BT 26.250 419.679 Td /F1 9.8 Tf [(In addition to data collected through the Intellicage hardware \(visits to each of the four corners\) general activity data was )] TJ ET BT 26.250 407.774 Td /F1 9.8 Tf [(collected through PsychoGenics Incs proprietary computer vision automated video scoring system. Our computer vision detects )] TJ ET BT 26.250 395.869 Td /F1 9.8 Tf [(and separates individual animals in the central area of the cage. As all mice in a cage had the same genotype we did not track )] TJ ET BT 26.250 383.964 Td /F1 9.8 Tf [(their identity across the whole session time, although the system was developed since to do so; accordingly, these data are )] TJ ET BT 26.250 372.060 Td /F1 9.8 Tf [(pooled across individuals within a cage. The primary measures currently implemented in this system are outlined below \(note )] TJ ET BT 26.250 360.155 Td /F1 9.8 Tf [(that animals within a corner are invisible to the camera and are accordingly not scored\):)] TJ ET BT 26.250 340.750 Td /F5 9.8 Tf [(Locomotion: )] TJ ET BT 82.069 340.750 Td /F1 9.8 Tf [(Measured as the distance travelled and normalized by the number of mice detected in each frame.)] TJ ET BT 26.250 321.345 Td /F5 9.8 Tf [(Immobility:)] TJ ET BT 72.835 321.345 Td /F1 9.8 Tf [( Measured as the time when an individual mouse was detected not moving \(maximum speed cutoff: 0.7 cm/s\).)] TJ ET BT 26.250 301.941 Td /F5 9.8 Tf [(Climbing:)] TJ ET BT 66.878 301.941 Td /F1 9.8 Tf [( Scored when a mouse was detected climbing on either of the climbing rods along the long wall of the cage or on the )] TJ ET BT 26.250 290.036 Td /F1 9.8 Tf [(climbing structure \(see Figure 1, left panel\).)] TJ ET BT 26.250 270.631 Td /F5 9.8 Tf [(Rearing:)] TJ ET BT 63.095 270.631 Td /F1 9.8 Tf [( Scored when an individual mouse was detected rearing against the walls of the apparatus.)] TJ ET BT 26.250 251.226 Td /F4 9.8 Tf [(Data analysis:)] TJ ET BT 26.250 231.822 Td /F1 9.8 Tf [(We analyzed data with mixed-design three-way ANOVA with SAS \(SAS Institute Inc.\) using Mixed Effect Models, based on )] TJ ET BT 26.250 219.917 Td /F1 9.8 Tf [(likelihood estimation. The models were fitted using the procedure PROC MIXED using genotype as the between-subjects )] TJ ET BT 26.250 208.012 Td /F1 9.8 Tf [(variable and light cycle and age as within-subjects variable. Significant interactions were followed by the simple effects analysis. )] TJ ET BT 26.250 196.107 Td /F1 9.8 Tf [(Where multiple interactions are present involving a given factor, simple effects analyses were conducted only for the highest )] TJ ET BT 26.250 184.203 Td /F1 9.8 Tf [(order interaction. An effect was considered significant if )] TJ ET BT 267.397 184.203 Td /F5 9.8 Tf [(p)] TJ ET BT 272.818 184.203 Td /F1 9.8 Tf [( < 0.05.)] TJ ET BT 26.250 147.600 Td /F4 12.0 Tf [(Results)] TJ ET BT 26.250 127.646 Td /F4 9.8 Tf [(Experiment 1. Analysis of R6/2 mice)] TJ ET BT 193.150 127.646 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 108.241 Td /F5 9.8 Tf [(Corner Entries: )] TJ ET BT 94.519 108.241 Td /F1 9.8 Tf [(R6/2 and WT mice did not exhibit differential frequency of corner entries. Both groups visited the corners equally )] TJ ET BT 26.250 96.336 Td /F1 9.8 Tf [(often, showed more visits during the dark light cycle, and reduced corner entries with repeated testing \(Figure 5A; Table I; Age )] TJ ET BT 26.250 84.432 Td /F1 9.8 Tf [(and Cycle main effects: )] TJ ET BT 130.292 84.432 Td /F5 9.8 Tf [(p)] TJ ET BT 135.713 84.432 Td /F1 9.8 Tf [(s < 0.05; simple effects analysis: )] TJ ET BT 279.048 84.432 Td /F5 9.8 Tf [(ps < )] TJ ET BT 300.459 84.432 Td /F1 9.8 Tf [(0)] TJ ET BT 305.880 84.432 Td /F5 9.8 Tf [(.)] TJ ET BT 308.591 84.432 Td /F1 9.8 Tf [(0001\). R6/2 mice showed slightly larger differences across the )] TJ ET BT 26.250 72.527 Td /F1 9.8 Tf [(day cycle although the differences between groups were not significant within each phase \(Cycle x Genotype interaction: )] TJ ET BT 548.665 72.527 Td /F5 9.8 Tf [(p )] TJ ET BT 556.796 72.527 Td /F1 9.8 Tf [(< )] TJ ET BT 26.250 60.622 Td /F1 9.8 Tf [(0.05; simple effects analysis: )] TJ ET BT 153.595 60.622 Td /F5 9.8 Tf [(p)] TJ ET BT 159.016 60.622 Td /F1 9.8 Tf [(s > 0.11\).)] TJ ET BT 26.250 41.217 Td /F5 9.8 Tf [(Licking:)] TJ ET BT 59.303 41.217 Td /F1 9.8 Tf [( Analysis of licking behavior pointed at increased thirst in R6/2 mice during the dark period. R6/2 mice licked more )] TJ ET Q q 15.000 26.932 577.500 750.068 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F4 9.8 Tf [(Experiment 2. )] TJ ET BT 92.901 767.476 Td /F1 9.8 Tf [(BACHD mice were evaluated longitudinally, with testing at 16, 24, 36, and 52 weeks of age. The subjects were a )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(cohort of 64 nave female mice, split evenly into four cages of 8 mice per genotype \(a total of eight cages\), each tested in a )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(single PhenoCube)] TJ ET BT 105.917 747.555 Td /F1 8.7 Tf [(TM)] TJ ET BT 118.432 743.667 Td /F1 9.8 Tf [(. In contrast to the R6/2 study, the animals were tested in a single long experimental session at each test )] TJ ET BT 26.250 731.762 Td /F1 9.8 Tf [(age, with the procedures employed otherwise as described for the R6/2 experiment. The timeline for these test sessions is )] TJ ET BT 26.250 719.857 Td /F1 9.8 Tf [(summarized in Figure 4, below. Note that, during the initial test at 16 weeks, the mice received an initial 24 hour period on the )] TJ ET BT 26.250 707.952 Td /F1 9.8 Tf [(simple Habituation protocol, to allow them to become accustomed to the cage environment, before switching to Alternation for )] TJ ET BT 26.250 696.048 Td /F1 9.8 Tf [(48 hours. In the later tests, the mice were placed immediately onto the Alternation protocol, with the corner contingency )] TJ ET BT 26.250 684.143 Td /F1 9.8 Tf [(reversing after 24 hours.)] TJ ET 0.965 0.965 0.965 rg 26.250 456.107 555.000 218.155 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 674.262 m 581.250 674.262 l 581.250 673.512 l 26.250 673.512 l f 26.250 456.107 m 581.250 456.107 l 581.250 456.857 l 26.250 456.857 l f q 450.000 0 0 173.250 35.250 491.262 cm /I6 Do Q q 35.250 467.357 537.000 17.905 re W n 0.271 0.267 0.267 rg BT 35.250 475.738 Td /F4 9.8 Tf [(Fig. 4: BACHD mice test procedure.)] TJ ET Q BT 26.250 439.083 Td /F4 9.8 Tf [(Computer vision:)] TJ ET BT 26.250 419.679 Td /F1 9.8 Tf [(In addition to data collected through the Intellicage hardware \(visits to each of the four corners\) general activity data was )] TJ ET BT 26.250 407.774 Td /F1 9.8 Tf [(collected through PsychoGenics Incs proprietary computer vision automated video scoring system. Our computer vision detects )] TJ ET BT 26.250 395.869 Td /F1 9.8 Tf [(and separates individual animals in the central area of the cage. As all mice in a cage had the same genotype we did not track )] TJ ET BT 26.250 383.964 Td /F1 9.8 Tf [(their identity across the whole session time, although the system was developed since to do so; accordingly, these data are )] TJ ET BT 26.250 372.060 Td /F1 9.8 Tf [(pooled across individuals within a cage. The primary measures currently implemented in this system are outlined below \(note )] TJ ET BT 26.250 360.155 Td /F1 9.8 Tf [(that animals within a corner are invisible to the camera and are accordingly not scored\):)] TJ ET BT 26.250 340.750 Td /F5 9.8 Tf [(Locomotion: )] TJ ET BT 82.069 340.750 Td /F1 9.8 Tf [(Measured as the distance travelled and normalized by the number of mice detected in each frame.)] TJ ET BT 26.250 321.345 Td /F5 9.8 Tf [(Immobility:)] TJ ET BT 72.835 321.345 Td /F1 9.8 Tf [( Measured as the time when an individual mouse was detected not moving \(maximum speed cutoff: 0.7 cm/s\).)] TJ ET BT 26.250 301.941 Td /F5 9.8 Tf [(Climbing:)] TJ ET BT 66.878 301.941 Td /F1 9.8 Tf [( Scored when a mouse was detected climbing on either of the climbing rods along the long wall of the cage or on the )] TJ ET BT 26.250 290.036 Td /F1 9.8 Tf [(climbing structure \(see Figure 1, left panel\).)] TJ ET BT 26.250 270.631 Td /F5 9.8 Tf [(Rearing:)] TJ ET BT 63.095 270.631 Td /F1 9.8 Tf [( Scored when an individual mouse was detected rearing against the walls of the apparatus.)] TJ ET BT 26.250 251.226 Td /F4 9.8 Tf [(Data analysis:)] TJ ET BT 26.250 231.822 Td /F1 9.8 Tf [(We analyzed data with mixed-design three-way ANOVA with SAS \(SAS Institute Inc.\) using Mixed Effect Models, based on )] TJ ET BT 26.250 219.917 Td /F1 9.8 Tf [(likelihood estimation. The models were fitted using the procedure PROC MIXED using genotype as the between-subjects )] TJ ET BT 26.250 208.012 Td /F1 9.8 Tf [(variable and light cycle and age as within-subjects variable. Significant interactions were followed by the simple effects analysis. )] TJ ET BT 26.250 196.107 Td /F1 9.8 Tf [(Where multiple interactions are present involving a given factor, simple effects analyses were conducted only for the highest )] TJ ET BT 26.250 184.203 Td /F1 9.8 Tf [(order interaction. An effect was considered significant if )] TJ ET BT 267.397 184.203 Td /F5 9.8 Tf [(p)] TJ ET BT 272.818 184.203 Td /F1 9.8 Tf [( < 0.05.)] TJ ET BT 26.250 147.600 Td /F4 12.0 Tf [(Results)] TJ ET BT 26.250 127.646 Td /F4 9.8 Tf [(Experiment 1. Analysis of R6/2 mice)] TJ ET BT 193.150 127.646 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 108.241 Td /F5 9.8 Tf [(Corner Entries: )] TJ ET BT 94.519 108.241 Td /F1 9.8 Tf [(R6/2 and WT mice did not exhibit differential frequency of corner entries. Both groups visited the corners equally )] TJ ET BT 26.250 96.336 Td /F1 9.8 Tf [(often, showed more visits during the dark light cycle, and reduced corner entries with repeated testing \(Figure 5A; Table I; Age )] TJ ET BT 26.250 84.432 Td /F1 9.8 Tf [(and Cycle main effects: )] TJ ET BT 130.292 84.432 Td /F5 9.8 Tf [(p)] TJ ET BT 135.713 84.432 Td /F1 9.8 Tf [(s < 0.05; simple effects analysis: )] TJ ET BT 279.048 84.432 Td /F5 9.8 Tf [(ps < )] TJ ET BT 300.459 84.432 Td /F1 9.8 Tf [(0)] TJ ET BT 305.880 84.432 Td /F5 9.8 Tf [(.)] TJ ET BT 308.591 84.432 Td /F1 9.8 Tf [(0001\). R6/2 mice showed slightly larger differences across the )] TJ ET BT 26.250 72.527 Td /F1 9.8 Tf [(day cycle although the differences between groups were not significant within each phase \(Cycle x Genotype interaction: )] TJ ET BT 548.665 72.527 Td /F5 9.8 Tf [(p )] TJ ET BT 556.796 72.527 Td /F1 9.8 Tf [(< )] TJ ET BT 26.250 60.622 Td /F1 9.8 Tf [(0.05; simple effects analysis: )] TJ ET BT 153.595 60.622 Td /F5 9.8 Tf [(p)] TJ ET BT 159.016 60.622 Td /F1 9.8 Tf [(s > 0.11\).)] TJ ET BT 26.250 41.217 Td /F5 9.8 Tf [(Licking:)] TJ ET BT 59.303 41.217 Td /F1 9.8 Tf [( Analysis of licking behavior pointed at increased thirst in R6/2 mice during the dark period. 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Surprisingly )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(the WT licked as frequently during the light as during the dark cycle \(Figure 5B; Table 1; Genotype x Age x Cycle interaction: )] TJ ET BT 565.444 755.571 Td /F5 9.8 Tf [(p )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(< 0.01; simple main effects: )] TJ ET BT 147.911 743.667 Td /F5 9.8 Tf [(p)] TJ ET BT 153.332 743.667 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET BT 26.250 724.262 Td /F5 9.8 Tf [(Repeats:)] TJ ET BT 65.269 724.262 Td /F1 9.8 Tf [( R6/2 mice re-entered visited corners more frequently than the WT controls, at all ages, which reflects the )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(perseverative responding R6/2 mouse behavior, which was \(although to a lesser extent\) also manifested in terms of their )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(alternation performance \(see below\). This effect appeared more pronounced during the light period \(Figure 5C; Table 1; )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(Genotype x Cycle interaction: )] TJ ET BT 156.305 688.548 Td /F5 9.8 Tf [(p )] TJ ET BT 164.437 688.548 Td /F1 9.8 Tf [(< 0.01; simple main effects: )] TJ ET BT 286.097 688.548 Td /F5 9.8 Tf [(p)] TJ ET BT 291.518 688.548 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET BT 26.250 669.143 Td /F5 9.8 Tf [(Alternations:)] TJ ET BT 80.441 669.143 Td /F1 9.8 Tf [( As mentioned above, perseverative responding R6/2 behavior was also apparent in the spatial alternation )] TJ ET BT 26.250 657.238 Td /F1 9.8 Tf [(performance; however genotypic differences were not as robust at older ages when compared to the deficits detected in )] TJ ET BT 26.250 645.333 Td /F1 9.8 Tf [(repeated behavior. R6/2 animals alternated less than WT mice between active corners, where water could be obtained, )] TJ ET BT 26.250 633.429 Td /F1 9.8 Tf [(although the differences were not significant at 9 weeks or at 10 weeks during the dark phase. Whereas WT mice alternated to )] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(the same extent during the light and dark cycle, R6/2 mice showed a further decrease in alternation during the light cycle phase )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(at 8 weeks of age \(Figure 5D; Table 1; Genotype x Age x Cycle interaction: p < 0.05; Simple Main Effects: )] TJ ET BT 484.988 609.619 Td /F5 9.8 Tf [(p)] TJ ET BT 490.409 609.619 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET BT 26.250 590.214 Td /F5 9.8 Tf [(Collected Rewards:)] TJ ET BT 110.773 590.214 Td /F1 9.8 Tf [( Despite the behavioral differences reported above R6/2 and WT mice had comparable levels of access to )] TJ ET BT 26.250 578.310 Td /F1 9.8 Tf [(water reinforcement earned through the experimental contingencies. The proportion of entries to an active corner in which mice )] TJ ET BT 26.250 566.405 Td /F1 9.8 Tf [(nosepoke and, likely, accessed the water reinforcement was similar between groups \(Figure 5E, Table 1\). There was a marginal )] TJ ET BT 26.250 554.500 Td /F1 9.8 Tf [(non significant increase in collected rewards with repeated testing \(Age main effect)] TJ ET BT 383.363 554.500 Td /F5 9.8 Tf [(: p)] TJ ET BT 394.205 554.500 Td /F1 9.8 Tf [( < 0.1\).)] TJ ET q 26.250 517.147 555.000 27.473 re W n 0.271 0.267 0.267 rg BT 26.250 533.630 Td /F1 9.8 Tf [(Table 1. Summary of the main effects and interaction following statistical analysis for the behavioral measures for Experiment 1 )] TJ ET BT 26.250 519.894 Td /F1 9.8 Tf [(\(R6/2\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 486.239 555.000 23.408 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 509.647 m 581.250 509.647 l 580.500 508.897 l 27.000 508.897 l f 581.250 509.647 m 581.250 486.239 l 580.500 486.239 l 580.500 508.897 l f 26.250 509.647 m 26.250 486.239 l 27.000 486.239 l 27.000 508.897 l f 0.271 0.267 0.267 rg BT 33.000 495.571 Td /F1 9.0 Tf [(Note: *: p<.05, **: p<.01, #: p<.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 356.214 555.000 130.025 re f 0.965 0.965 0.965 rg 27.000 436.370 124.091 49.119 re f 0.267 0.267 0.267 rg 26.625 485.114 124.466 0.750 re f 26.625 473.209 0.750 12.655 re f 26.625 460.930 0.750 12.280 re f 26.625 448.650 0.750 12.280 re f 26.625 435.995 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 475.965 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 151.091 473.209 429.409 12.280 re f 0.267 0.267 0.267 rg 151.091 485.114 85.941 0.750 re f 237.032 485.114 64.374 0.750 re f 301.407 485.114 71.640 0.750 re f 373.047 485.114 64.374 0.750 re f 437.421 485.114 71.640 0.750 re f 509.061 485.114 71.814 0.750 re f 580.125 472.834 0.750 13.030 re f 0.271 0.267 0.267 rg BT 155.591 475.965 Td /F4 9.8 Tf [(R6/2)] TJ ET 0.965 0.965 0.965 rg 151.091 460.930 85.941 12.280 re f 0.267 0.267 0.267 rg 151.091 472.834 85.941 0.750 re f 0.271 0.267 0.267 rg BT 155.591 463.686 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 237.032 460.930 64.374 12.280 re f 0.267 0.267 0.267 rg 237.032 472.834 64.374 0.750 re f 0.271 0.267 0.267 rg BT 241.532 463.686 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 301.407 460.930 71.640 12.280 re f 0.267 0.267 0.267 rg 301.407 472.834 71.640 0.750 re f 0.271 0.267 0.267 rg BT 305.907 463.686 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 373.047 460.930 64.374 12.280 re f 0.267 0.267 0.267 rg 373.047 472.834 64.374 0.750 re f 0.271 0.267 0.267 rg BT 377.547 463.686 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 437.421 460.930 71.640 12.280 re f 0.267 0.267 0.267 rg 437.421 472.834 71.640 0.750 re f 0.271 0.267 0.267 rg BT 441.921 463.686 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 509.061 460.930 71.439 12.280 re f 0.267 0.267 0.267 rg 509.061 472.834 71.814 0.750 re f 580.125 460.555 0.750 13.030 re f 0.271 0.267 0.267 rg BT 513.561 463.686 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 151.091 448.650 85.941 12.280 re f 0.267 0.267 0.267 rg 151.091 460.555 85.941 0.750 re f 0.271 0.267 0.267 rg BT 155.591 451.406 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 237.032 448.650 64.374 12.280 re f 0.267 0.267 0.267 rg 237.032 460.555 64.374 0.750 re f 0.271 0.267 0.267 rg BT 241.532 451.406 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 301.407 448.650 71.640 12.280 re f 0.267 0.267 0.267 rg 301.407 460.555 71.640 0.750 re f 0.271 0.267 0.267 rg BT 305.907 451.406 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 373.047 448.650 64.374 12.280 re f 0.267 0.267 0.267 rg 373.047 460.555 64.374 0.750 re f 0.965 0.965 0.965 rg 437.421 448.650 71.640 12.280 re f 0.267 0.267 0.267 rg 437.421 460.555 71.640 0.750 re f 0.965 0.965 0.965 rg 509.061 448.650 71.439 12.280 re f 0.267 0.267 0.267 rg 509.061 460.555 71.814 0.750 re f 580.125 448.275 0.750 13.030 re f 0.965 0.965 0.965 rg 151.091 436.370 85.941 12.280 re f 0.267 0.267 0.267 rg 151.091 448.275 85.941 0.750 re f 0.271 0.267 0.267 rg BT 155.591 439.126 Td /F4 9.8 Tf [(F\(1,38\))] TJ ET 0.965 0.965 0.965 rg 237.032 436.370 64.374 12.280 re f 0.267 0.267 0.267 rg 237.032 448.275 64.374 0.750 re f 0.271 0.267 0.267 rg BT 241.532 439.126 Td /F4 9.8 Tf [(F\(3,56\))] TJ ET 0.965 0.965 0.965 rg 301.407 436.370 71.640 12.280 re f 0.267 0.267 0.267 rg 301.407 448.275 71.640 0.750 re f 0.271 0.267 0.267 rg BT 305.907 439.126 Td /F4 9.8 Tf [(F\(1,38\))] TJ ET 0.965 0.965 0.965 rg 373.047 436.370 64.374 12.280 re f 0.267 0.267 0.267 rg 373.047 448.275 64.374 0.750 re f 0.271 0.267 0.267 rg BT 377.547 439.126 Td /F4 9.8 Tf [(F\(3,56\))] TJ ET 0.965 0.965 0.965 rg 437.421 436.370 71.640 12.280 re f 0.267 0.267 0.267 rg 437.421 448.275 71.640 0.750 re f 0.271 0.267 0.267 rg BT 441.921 439.126 Td /F4 9.8 Tf [(F\(1,38\))] TJ ET 0.965 0.965 0.965 rg 509.061 436.370 71.439 12.280 re f 0.267 0.267 0.267 rg 509.061 448.275 71.814 0.750 re f 580.125 435.995 0.750 13.030 re f 0.271 0.267 0.267 rg BT 513.561 439.126 Td /F4 9.8 Tf [(F\(6,66\))] TJ ET 0.267 0.267 0.267 rg 26.625 435.995 124.841 0.750 re f 26.625 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 426.140 Td /F4 9.8 Tf [(Corner Entries)] TJ ET 0.267 0.267 0.267 rg 150.716 435.995 86.691 0.750 re f 150.716 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 426.140 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 236.657 435.995 65.124 0.750 re f 236.657 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 426.140 Td /F1 9.8 Tf [(4.13*)] TJ ET 0.267 0.267 0.267 rg 301.032 435.995 72.390 0.750 re f 301.032 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 426.140 Td /F1 9.8 Tf [(96.72#)] TJ ET 0.267 0.267 0.267 rg 372.672 435.995 65.124 0.750 re f 372.672 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 426.140 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 435.995 72.390 0.750 re f 437.046 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 426.140 Td /F1 9.8 Tf [(4.19*)] TJ ET 0.267 0.267 0.267 rg 508.686 435.995 72.189 0.750 re f 508.686 420.114 0.750 16.631 re f 580.125 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 426.140 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 420.114 124.841 0.750 re f 26.625 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 410.259 Td /F4 9.8 Tf [(Licking)] TJ ET 0.267 0.267 0.267 rg 150.716 420.114 86.691 0.750 re f 150.716 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 410.259 Td /F1 9.8 Tf [(17.46#)] TJ ET 0.267 0.267 0.267 rg 236.657 420.114 65.124 0.750 re f 236.657 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 410.259 Td /F1 9.8 Tf [(3.67*)] TJ ET 0.267 0.267 0.267 rg 301.032 420.114 72.390 0.750 re f 301.032 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 410.259 Td /F1 9.8 Tf [(149.36#)] TJ ET 0.267 0.267 0.267 rg 372.672 420.114 65.124 0.750 re f 372.672 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 410.259 Td /F1 9.8 Tf [(5.26**)] TJ ET 0.267 0.267 0.267 rg 437.046 420.114 72.390 0.750 re f 437.046 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 410.259 Td /F1 9.8 Tf [(140.32#)] TJ ET 0.267 0.267 0.267 rg 508.686 420.114 72.189 0.750 re f 508.686 404.233 0.750 16.631 re f 580.125 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 410.259 Td /F1 9.8 Tf [(4.14**)] TJ ET 0.267 0.267 0.267 rg 26.625 404.233 124.841 0.750 re f 26.625 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 394.378 Td /F4 9.8 Tf [(Repeats)] TJ ET 0.267 0.267 0.267 rg 150.716 404.233 86.691 0.750 re f 150.716 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 394.378 Td /F1 9.8 Tf [(83.97#)] TJ ET 0.267 0.267 0.267 rg 236.657 404.233 65.124 0.750 re f 236.657 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 301.032 404.233 72.390 0.750 re f 301.032 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 372.672 404.233 65.124 0.750 re f 372.672 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 404.233 72.390 0.750 re f 437.046 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 394.378 Td /F1 9.8 Tf [(21.80#)] TJ ET 0.267 0.267 0.267 rg 508.686 404.233 72.189 0.750 re f 508.686 388.351 0.750 16.631 re f 580.125 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 388.351 124.841 0.750 re f 26.625 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 378.496 Td /F4 9.8 Tf [(Alternations)] TJ ET 0.267 0.267 0.267 rg 150.716 388.351 86.691 0.750 re f 150.716 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 378.496 Td /F1 9.8 Tf [(33.25#)] TJ ET 0.267 0.267 0.267 rg 236.657 388.351 65.124 0.750 re f 236.657 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 378.496 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 301.032 388.351 72.390 0.750 re f 301.032 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 378.496 Td /F1 9.8 Tf [(5.66*)] TJ ET 0.267 0.267 0.267 rg 372.672 388.351 65.124 0.750 re f 372.672 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 378.496 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 388.351 72.390 0.750 re f 437.046 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 378.496 Td /F1 9.8 Tf [(7.30*)] TJ ET 0.267 0.267 0.267 rg 508.686 388.351 72.189 0.750 re f 508.686 372.470 0.750 16.631 re f 580.125 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 378.496 Td /F1 9.8 Tf [(2.39*)] TJ ET 0.267 0.267 0.267 rg 26.625 372.470 124.841 0.750 re f 26.625 356.589 124.841 0.750 re f 26.625 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 362.615 Td /F4 9.8 Tf [(Collected Rewards)] TJ ET 0.267 0.267 0.267 rg 150.716 372.470 86.691 0.750 re f 150.716 356.589 86.691 0.750 re f 150.716 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 236.657 372.470 65.124 0.750 re f 236.657 356.589 65.124 0.750 re f 236.657 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 301.032 372.470 72.390 0.750 re f 301.032 356.589 72.390 0.750 re f 301.032 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 372.672 372.470 65.124 0.750 re f 372.672 356.589 65.124 0.750 re f 372.672 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 372.470 72.390 0.750 re f 437.046 356.589 72.390 0.750 re f 437.046 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 508.686 372.470 72.189 0.750 re f 508.686 356.589 72.189 0.750 re f 508.686 356.589 0.750 16.631 re f 580.125 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.965 0.965 0.965 rg 26.250 -154.536 555.000 465.750 re f 0.267 0.267 0.267 rg 26.250 311.214 m 581.250 311.214 l 581.250 310.464 l 26.250 310.464 l f q 321.000 0 0 450.000 35.250 -148.536 cm /I7 Do Q q 35.250 -154.536 537.000 0.000 re W n Q Q q 15.000 -154.536 577.500 931.536 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(frequently than the WT controls during the dark phase, although the effect was not significant at 10 weeks of age. Surprisingly )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(the WT licked as frequently during the light as during the dark cycle \(Figure 5B; Table 1; Genotype x Age x Cycle interaction: )] TJ ET BT 565.444 755.571 Td /F5 9.8 Tf [(p )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(< 0.01; simple main effects: )] TJ ET BT 147.911 743.667 Td /F5 9.8 Tf [(p)] TJ ET BT 153.332 743.667 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET BT 26.250 724.262 Td /F5 9.8 Tf [(Repeats:)] TJ ET BT 65.269 724.262 Td /F1 9.8 Tf [( R6/2 mice re-entered visited corners more frequently than the WT controls, at all ages, which reflects the )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(perseverative responding R6/2 mouse behavior, which was \(although to a lesser extent\) also manifested in terms of their )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(alternation performance \(see below\). This effect appeared more pronounced during the light period \(Figure 5C; Table 1; )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(Genotype x Cycle interaction: )] TJ ET BT 156.305 688.548 Td /F5 9.8 Tf [(p )] TJ ET BT 164.437 688.548 Td /F1 9.8 Tf [(< 0.01; simple main effects: )] TJ ET BT 286.097 688.548 Td /F5 9.8 Tf [(p)] TJ ET BT 291.518 688.548 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET BT 26.250 669.143 Td /F5 9.8 Tf [(Alternations:)] TJ ET BT 80.441 669.143 Td /F1 9.8 Tf [( As mentioned above, perseverative responding R6/2 behavior was also apparent in the spatial alternation )] TJ ET BT 26.250 657.238 Td /F1 9.8 Tf [(performance; however genotypic differences were not as robust at older ages when compared to the deficits detected in )] TJ ET BT 26.250 645.333 Td /F1 9.8 Tf [(repeated behavior. R6/2 animals alternated less than WT mice between active corners, where water could be obtained, )] TJ ET BT 26.250 633.429 Td /F1 9.8 Tf [(although the differences were not significant at 9 weeks or at 10 weeks during the dark phase. Whereas WT mice alternated to )] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(the same extent during the light and dark cycle, R6/2 mice showed a further decrease in alternation during the light cycle phase )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(at 8 weeks of age \(Figure 5D; Table 1; Genotype x Age x Cycle interaction: p < 0.05; Simple Main Effects: )] TJ ET BT 484.988 609.619 Td /F5 9.8 Tf [(p)] TJ ET BT 490.409 609.619 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET BT 26.250 590.214 Td /F5 9.8 Tf [(Collected Rewards:)] TJ ET BT 110.773 590.214 Td /F1 9.8 Tf [( Despite the behavioral differences reported above R6/2 and WT mice had comparable levels of access to )] TJ ET BT 26.250 578.310 Td /F1 9.8 Tf [(water reinforcement earned through the experimental contingencies. The proportion of entries to an active corner in which mice )] TJ ET BT 26.250 566.405 Td /F1 9.8 Tf [(nosepoke and, likely, accessed the water reinforcement was similar between groups \(Figure 5E, Table 1\). There was a marginal )] TJ ET BT 26.250 554.500 Td /F1 9.8 Tf [(non significant increase in collected rewards with repeated testing \(Age main effect)] TJ ET BT 383.363 554.500 Td /F5 9.8 Tf [(: p)] TJ ET BT 394.205 554.500 Td /F1 9.8 Tf [( < 0.1\).)] TJ ET q 26.250 517.147 555.000 27.473 re W n 0.271 0.267 0.267 rg BT 26.250 533.630 Td /F1 9.8 Tf [(Table 1. Summary of the main effects and interaction following statistical analysis for the behavioral measures for Experiment 1 )] TJ ET BT 26.250 519.894 Td /F1 9.8 Tf [(\(R6/2\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 486.239 555.000 23.408 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 509.647 m 581.250 509.647 l 580.500 508.897 l 27.000 508.897 l f 581.250 509.647 m 581.250 486.239 l 580.500 486.239 l 580.500 508.897 l f 26.250 509.647 m 26.250 486.239 l 27.000 486.239 l 27.000 508.897 l f 0.271 0.267 0.267 rg BT 33.000 495.571 Td /F1 9.0 Tf [(Note: *: p<.05, **: p<.01, #: p<.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 356.214 555.000 130.025 re f 0.965 0.965 0.965 rg 27.000 436.370 124.091 49.119 re f 0.267 0.267 0.267 rg 26.625 485.114 124.466 0.750 re f 26.625 473.209 0.750 12.655 re f 26.625 460.930 0.750 12.280 re f 26.625 448.650 0.750 12.280 re f 26.625 435.995 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 475.965 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 151.091 473.209 429.409 12.280 re f 0.267 0.267 0.267 rg 151.091 485.114 85.941 0.750 re f 237.032 485.114 64.374 0.750 re f 301.407 485.114 71.640 0.750 re f 373.047 485.114 64.374 0.750 re f 437.421 485.114 71.640 0.750 re f 509.061 485.114 71.814 0.750 re f 580.125 472.834 0.750 13.030 re f 0.271 0.267 0.267 rg BT 155.591 475.965 Td /F4 9.8 Tf [(R6/2)] TJ ET 0.965 0.965 0.965 rg 151.091 460.930 85.941 12.280 re f 0.267 0.267 0.267 rg 151.091 472.834 85.941 0.750 re f 0.271 0.267 0.267 rg BT 155.591 463.686 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 237.032 460.930 64.374 12.280 re f 0.267 0.267 0.267 rg 237.032 472.834 64.374 0.750 re f 0.271 0.267 0.267 rg BT 241.532 463.686 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 301.407 460.930 71.640 12.280 re f 0.267 0.267 0.267 rg 301.407 472.834 71.640 0.750 re f 0.271 0.267 0.267 rg BT 305.907 463.686 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 373.047 460.930 64.374 12.280 re f 0.267 0.267 0.267 rg 373.047 472.834 64.374 0.750 re f 0.271 0.267 0.267 rg BT 377.547 463.686 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 437.421 460.930 71.640 12.280 re f 0.267 0.267 0.267 rg 437.421 472.834 71.640 0.750 re f 0.271 0.267 0.267 rg BT 441.921 463.686 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 509.061 460.930 71.439 12.280 re f 0.267 0.267 0.267 rg 509.061 472.834 71.814 0.750 re f 580.125 460.555 0.750 13.030 re f 0.271 0.267 0.267 rg BT 513.561 463.686 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 151.091 448.650 85.941 12.280 re f 0.267 0.267 0.267 rg 151.091 460.555 85.941 0.750 re f 0.271 0.267 0.267 rg BT 155.591 451.406 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 237.032 448.650 64.374 12.280 re f 0.267 0.267 0.267 rg 237.032 460.555 64.374 0.750 re f 0.271 0.267 0.267 rg BT 241.532 451.406 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 301.407 448.650 71.640 12.280 re f 0.267 0.267 0.267 rg 301.407 460.555 71.640 0.750 re f 0.271 0.267 0.267 rg BT 305.907 451.406 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 373.047 448.650 64.374 12.280 re f 0.267 0.267 0.267 rg 373.047 460.555 64.374 0.750 re f 0.965 0.965 0.965 rg 437.421 448.650 71.640 12.280 re f 0.267 0.267 0.267 rg 437.421 460.555 71.640 0.750 re f 0.965 0.965 0.965 rg 509.061 448.650 71.439 12.280 re f 0.267 0.267 0.267 rg 509.061 460.555 71.814 0.750 re f 580.125 448.275 0.750 13.030 re f 0.965 0.965 0.965 rg 151.091 436.370 85.941 12.280 re f 0.267 0.267 0.267 rg 151.091 448.275 85.941 0.750 re f 0.271 0.267 0.267 rg BT 155.591 439.126 Td /F4 9.8 Tf [(F\(1,38\))] TJ ET 0.965 0.965 0.965 rg 237.032 436.370 64.374 12.280 re f 0.267 0.267 0.267 rg 237.032 448.275 64.374 0.750 re f 0.271 0.267 0.267 rg BT 241.532 439.126 Td /F4 9.8 Tf [(F\(3,56\))] TJ ET 0.965 0.965 0.965 rg 301.407 436.370 71.640 12.280 re f 0.267 0.267 0.267 rg 301.407 448.275 71.640 0.750 re f 0.271 0.267 0.267 rg BT 305.907 439.126 Td /F4 9.8 Tf [(F\(1,38\))] TJ ET 0.965 0.965 0.965 rg 373.047 436.370 64.374 12.280 re f 0.267 0.267 0.267 rg 373.047 448.275 64.374 0.750 re f 0.271 0.267 0.267 rg BT 377.547 439.126 Td /F4 9.8 Tf [(F\(3,56\))] TJ ET 0.965 0.965 0.965 rg 437.421 436.370 71.640 12.280 re f 0.267 0.267 0.267 rg 437.421 448.275 71.640 0.750 re f 0.271 0.267 0.267 rg BT 441.921 439.126 Td /F4 9.8 Tf [(F\(1,38\))] TJ ET 0.965 0.965 0.965 rg 509.061 436.370 71.439 12.280 re f 0.267 0.267 0.267 rg 509.061 448.275 71.814 0.750 re f 580.125 435.995 0.750 13.030 re f 0.271 0.267 0.267 rg BT 513.561 439.126 Td /F4 9.8 Tf [(F\(6,66\))] TJ ET 0.267 0.267 0.267 rg 26.625 435.995 124.841 0.750 re f 26.625 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 426.140 Td /F4 9.8 Tf [(Corner Entries)] TJ ET 0.267 0.267 0.267 rg 150.716 435.995 86.691 0.750 re f 150.716 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 426.140 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 236.657 435.995 65.124 0.750 re f 236.657 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 426.140 Td /F1 9.8 Tf [(4.13*)] TJ ET 0.267 0.267 0.267 rg 301.032 435.995 72.390 0.750 re f 301.032 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 426.140 Td /F1 9.8 Tf [(96.72#)] TJ ET 0.267 0.267 0.267 rg 372.672 435.995 65.124 0.750 re f 372.672 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 426.140 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 435.995 72.390 0.750 re f 437.046 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 426.140 Td /F1 9.8 Tf [(4.19*)] TJ ET 0.267 0.267 0.267 rg 508.686 435.995 72.189 0.750 re f 508.686 420.114 0.750 16.631 re f 580.125 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 426.140 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 420.114 124.841 0.750 re f 26.625 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 410.259 Td /F4 9.8 Tf [(Licking)] TJ ET 0.267 0.267 0.267 rg 150.716 420.114 86.691 0.750 re f 150.716 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 410.259 Td /F1 9.8 Tf [(17.46#)] TJ ET 0.267 0.267 0.267 rg 236.657 420.114 65.124 0.750 re f 236.657 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 410.259 Td /F1 9.8 Tf [(3.67*)] TJ ET 0.267 0.267 0.267 rg 301.032 420.114 72.390 0.750 re f 301.032 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 410.259 Td /F1 9.8 Tf [(149.36#)] TJ ET 0.267 0.267 0.267 rg 372.672 420.114 65.124 0.750 re f 372.672 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 410.259 Td /F1 9.8 Tf [(5.26**)] TJ ET 0.267 0.267 0.267 rg 437.046 420.114 72.390 0.750 re f 437.046 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 410.259 Td /F1 9.8 Tf [(140.32#)] TJ ET 0.267 0.267 0.267 rg 508.686 420.114 72.189 0.750 re f 508.686 404.233 0.750 16.631 re f 580.125 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 410.259 Td /F1 9.8 Tf [(4.14**)] TJ ET 0.267 0.267 0.267 rg 26.625 404.233 124.841 0.750 re f 26.625 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 394.378 Td /F4 9.8 Tf [(Repeats)] TJ ET 0.267 0.267 0.267 rg 150.716 404.233 86.691 0.750 re f 150.716 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 394.378 Td /F1 9.8 Tf [(83.97#)] TJ ET 0.267 0.267 0.267 rg 236.657 404.233 65.124 0.750 re f 236.657 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 301.032 404.233 72.390 0.750 re f 301.032 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 372.672 404.233 65.124 0.750 re f 372.672 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 404.233 72.390 0.750 re f 437.046 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 394.378 Td /F1 9.8 Tf [(21.80#)] TJ ET 0.267 0.267 0.267 rg 508.686 404.233 72.189 0.750 re f 508.686 388.351 0.750 16.631 re f 580.125 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 388.351 124.841 0.750 re f 26.625 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 378.496 Td /F4 9.8 Tf [(Alternations)] TJ ET 0.267 0.267 0.267 rg 150.716 388.351 86.691 0.750 re f 150.716 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 378.496 Td /F1 9.8 Tf [(33.25#)] TJ ET 0.267 0.267 0.267 rg 236.657 388.351 65.124 0.750 re f 236.657 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 378.496 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 301.032 388.351 72.390 0.750 re f 301.032 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 378.496 Td /F1 9.8 Tf [(5.66*)] TJ ET 0.267 0.267 0.267 rg 372.672 388.351 65.124 0.750 re f 372.672 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 378.496 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 388.351 72.390 0.750 re f 437.046 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 378.496 Td /F1 9.8 Tf [(7.30*)] TJ ET 0.267 0.267 0.267 rg 508.686 388.351 72.189 0.750 re f 508.686 372.470 0.750 16.631 re f 580.125 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 378.496 Td /F1 9.8 Tf [(2.39*)] TJ ET 0.267 0.267 0.267 rg 26.625 372.470 124.841 0.750 re f 26.625 356.589 124.841 0.750 re f 26.625 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 362.615 Td /F4 9.8 Tf [(Collected Rewards)] TJ ET 0.267 0.267 0.267 rg 150.716 372.470 86.691 0.750 re f 150.716 356.589 86.691 0.750 re f 150.716 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 236.657 372.470 65.124 0.750 re f 236.657 356.589 65.124 0.750 re f 236.657 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 301.032 372.470 72.390 0.750 re f 301.032 356.589 72.390 0.750 re f 301.032 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 372.672 372.470 65.124 0.750 re f 372.672 356.589 65.124 0.750 re f 372.672 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 372.470 72.390 0.750 re f 437.046 356.589 72.390 0.750 re f 437.046 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 508.686 372.470 72.189 0.750 re f 508.686 356.589 72.189 0.750 re f 508.686 356.589 0.750 16.631 re f 580.125 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.965 0.965 0.965 rg 26.250 -154.536 555.000 465.750 re f 0.267 0.267 0.267 rg 26.250 311.214 m 581.250 311.214 l 581.250 310.464 l 26.250 310.464 l f q 321.000 0 0 450.000 35.250 -148.536 cm /I7 Do Q q 35.250 -154.536 537.000 0.000 re W n Q Q q 15.000 -154.536 577.500 931.536 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(frequently than the WT controls during the dark phase, although the effect was not significant at 10 weeks of age. Surprisingly )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(the WT licked as frequently during the light as during the dark cycle \(Figure 5B; Table 1; Genotype x Age x Cycle interaction: )] TJ ET BT 565.444 755.571 Td /F5 9.8 Tf [(p )] TJ ET BT 26.250 743.667 Td /F1 9.8 Tf [(< 0.01; simple main effects: )] TJ ET BT 147.911 743.667 Td /F5 9.8 Tf [(p)] TJ ET BT 153.332 743.667 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET BT 26.250 724.262 Td /F5 9.8 Tf [(Repeats:)] TJ ET BT 65.269 724.262 Td /F1 9.8 Tf [( R6/2 mice re-entered visited corners more frequently than the WT controls, at all ages, which reflects the )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(perseverative responding R6/2 mouse behavior, which was \(although to a lesser extent\) also manifested in terms of their )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(alternation performance \(see below\). This effect appeared more pronounced during the light period \(Figure 5C; Table 1; )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(Genotype x Cycle interaction: )] TJ ET BT 156.305 688.548 Td /F5 9.8 Tf [(p )] TJ ET BT 164.437 688.548 Td /F1 9.8 Tf [(< 0.01; simple main effects: )] TJ ET BT 286.097 688.548 Td /F5 9.8 Tf [(p)] TJ ET BT 291.518 688.548 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET BT 26.250 669.143 Td /F5 9.8 Tf [(Alternations:)] TJ ET BT 80.441 669.143 Td /F1 9.8 Tf [( As mentioned above, perseverative responding R6/2 behavior was also apparent in the spatial alternation )] TJ ET BT 26.250 657.238 Td /F1 9.8 Tf [(performance; however genotypic differences were not as robust at older ages when compared to the deficits detected in )] TJ ET BT 26.250 645.333 Td /F1 9.8 Tf [(repeated behavior. R6/2 animals alternated less than WT mice between active corners, where water could be obtained, )] TJ ET BT 26.250 633.429 Td /F1 9.8 Tf [(although the differences were not significant at 9 weeks or at 10 weeks during the dark phase. Whereas WT mice alternated to )] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(the same extent during the light and dark cycle, R6/2 mice showed a further decrease in alternation during the light cycle phase )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(at 8 weeks of age \(Figure 5D; Table 1; Genotype x Age x Cycle interaction: p < 0.05; Simple Main Effects: )] TJ ET BT 484.988 609.619 Td /F5 9.8 Tf [(p)] TJ ET BT 490.409 609.619 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET BT 26.250 590.214 Td /F5 9.8 Tf [(Collected Rewards:)] TJ ET BT 110.773 590.214 Td /F1 9.8 Tf [( Despite the behavioral differences reported above R6/2 and WT mice had comparable levels of access to )] TJ ET BT 26.250 578.310 Td /F1 9.8 Tf [(water reinforcement earned through the experimental contingencies. The proportion of entries to an active corner in which mice )] TJ ET BT 26.250 566.405 Td /F1 9.8 Tf [(nosepoke and, likely, accessed the water reinforcement was similar between groups \(Figure 5E, Table 1\). There was a marginal )] TJ ET BT 26.250 554.500 Td /F1 9.8 Tf [(non significant increase in collected rewards with repeated testing \(Age main effect)] TJ ET BT 383.363 554.500 Td /F5 9.8 Tf [(: p)] TJ ET BT 394.205 554.500 Td /F1 9.8 Tf [( < 0.1\).)] TJ ET q 26.250 517.147 555.000 27.473 re W n 0.271 0.267 0.267 rg BT 26.250 533.630 Td /F1 9.8 Tf [(Table 1. Summary of the main effects and interaction following statistical analysis for the behavioral measures for Experiment 1 )] TJ ET BT 26.250 519.894 Td /F1 9.8 Tf [(\(R6/2\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 486.239 555.000 23.408 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 509.647 m 581.250 509.647 l 580.500 508.897 l 27.000 508.897 l f 581.250 509.647 m 581.250 486.239 l 580.500 486.239 l 580.500 508.897 l f 26.250 509.647 m 26.250 486.239 l 27.000 486.239 l 27.000 508.897 l f 0.271 0.267 0.267 rg BT 33.000 495.571 Td /F1 9.0 Tf [(Note: *: p<.05, **: p<.01, #: p<.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 356.214 555.000 130.025 re f 0.965 0.965 0.965 rg 27.000 436.370 124.091 49.119 re f 0.267 0.267 0.267 rg 26.625 485.114 124.466 0.750 re f 26.625 473.209 0.750 12.655 re f 26.625 460.930 0.750 12.280 re f 26.625 448.650 0.750 12.280 re f 26.625 435.995 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 475.965 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 151.091 473.209 429.409 12.280 re f 0.267 0.267 0.267 rg 151.091 485.114 85.941 0.750 re f 237.032 485.114 64.374 0.750 re f 301.407 485.114 71.640 0.750 re f 373.047 485.114 64.374 0.750 re f 437.421 485.114 71.640 0.750 re f 509.061 485.114 71.814 0.750 re f 580.125 472.834 0.750 13.030 re f 0.271 0.267 0.267 rg BT 155.591 475.965 Td /F4 9.8 Tf [(R6/2)] TJ ET 0.965 0.965 0.965 rg 151.091 460.930 85.941 12.280 re f 0.267 0.267 0.267 rg 151.091 472.834 85.941 0.750 re f 0.271 0.267 0.267 rg BT 155.591 463.686 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 237.032 460.930 64.374 12.280 re f 0.267 0.267 0.267 rg 237.032 472.834 64.374 0.750 re f 0.271 0.267 0.267 rg BT 241.532 463.686 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 301.407 460.930 71.640 12.280 re f 0.267 0.267 0.267 rg 301.407 472.834 71.640 0.750 re f 0.271 0.267 0.267 rg BT 305.907 463.686 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 373.047 460.930 64.374 12.280 re f 0.267 0.267 0.267 rg 373.047 472.834 64.374 0.750 re f 0.271 0.267 0.267 rg BT 377.547 463.686 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 437.421 460.930 71.640 12.280 re f 0.267 0.267 0.267 rg 437.421 472.834 71.640 0.750 re f 0.271 0.267 0.267 rg BT 441.921 463.686 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 509.061 460.930 71.439 12.280 re f 0.267 0.267 0.267 rg 509.061 472.834 71.814 0.750 re f 580.125 460.555 0.750 13.030 re f 0.271 0.267 0.267 rg BT 513.561 463.686 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 151.091 448.650 85.941 12.280 re f 0.267 0.267 0.267 rg 151.091 460.555 85.941 0.750 re f 0.271 0.267 0.267 rg BT 155.591 451.406 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 237.032 448.650 64.374 12.280 re f 0.267 0.267 0.267 rg 237.032 460.555 64.374 0.750 re f 0.271 0.267 0.267 rg BT 241.532 451.406 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 301.407 448.650 71.640 12.280 re f 0.267 0.267 0.267 rg 301.407 460.555 71.640 0.750 re f 0.271 0.267 0.267 rg BT 305.907 451.406 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 373.047 448.650 64.374 12.280 re f 0.267 0.267 0.267 rg 373.047 460.555 64.374 0.750 re f 0.965 0.965 0.965 rg 437.421 448.650 71.640 12.280 re f 0.267 0.267 0.267 rg 437.421 460.555 71.640 0.750 re f 0.965 0.965 0.965 rg 509.061 448.650 71.439 12.280 re f 0.267 0.267 0.267 rg 509.061 460.555 71.814 0.750 re f 580.125 448.275 0.750 13.030 re f 0.965 0.965 0.965 rg 151.091 436.370 85.941 12.280 re f 0.267 0.267 0.267 rg 151.091 448.275 85.941 0.750 re f 0.271 0.267 0.267 rg BT 155.591 439.126 Td /F4 9.8 Tf [(F\(1,38\))] TJ ET 0.965 0.965 0.965 rg 237.032 436.370 64.374 12.280 re f 0.267 0.267 0.267 rg 237.032 448.275 64.374 0.750 re f 0.271 0.267 0.267 rg BT 241.532 439.126 Td /F4 9.8 Tf [(F\(3,56\))] TJ ET 0.965 0.965 0.965 rg 301.407 436.370 71.640 12.280 re f 0.267 0.267 0.267 rg 301.407 448.275 71.640 0.750 re f 0.271 0.267 0.267 rg BT 305.907 439.126 Td /F4 9.8 Tf [(F\(1,38\))] TJ ET 0.965 0.965 0.965 rg 373.047 436.370 64.374 12.280 re f 0.267 0.267 0.267 rg 373.047 448.275 64.374 0.750 re f 0.271 0.267 0.267 rg BT 377.547 439.126 Td /F4 9.8 Tf [(F\(3,56\))] TJ ET 0.965 0.965 0.965 rg 437.421 436.370 71.640 12.280 re f 0.267 0.267 0.267 rg 437.421 448.275 71.640 0.750 re f 0.271 0.267 0.267 rg BT 441.921 439.126 Td /F4 9.8 Tf [(F\(1,38\))] TJ ET 0.965 0.965 0.965 rg 509.061 436.370 71.439 12.280 re f 0.267 0.267 0.267 rg 509.061 448.275 71.814 0.750 re f 580.125 435.995 0.750 13.030 re f 0.271 0.267 0.267 rg BT 513.561 439.126 Td /F4 9.8 Tf [(F\(6,66\))] TJ ET 0.267 0.267 0.267 rg 26.625 435.995 124.841 0.750 re f 26.625 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 426.140 Td /F4 9.8 Tf [(Corner Entries)] TJ ET 0.267 0.267 0.267 rg 150.716 435.995 86.691 0.750 re f 150.716 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 426.140 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 236.657 435.995 65.124 0.750 re f 236.657 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 426.140 Td /F1 9.8 Tf [(4.13*)] TJ ET 0.267 0.267 0.267 rg 301.032 435.995 72.390 0.750 re f 301.032 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 426.140 Td /F1 9.8 Tf [(96.72#)] TJ ET 0.267 0.267 0.267 rg 372.672 435.995 65.124 0.750 re f 372.672 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 426.140 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 435.995 72.390 0.750 re f 437.046 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 426.140 Td /F1 9.8 Tf [(4.19*)] TJ ET 0.267 0.267 0.267 rg 508.686 435.995 72.189 0.750 re f 508.686 420.114 0.750 16.631 re f 580.125 420.114 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 426.140 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 420.114 124.841 0.750 re f 26.625 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 410.259 Td /F4 9.8 Tf [(Licking)] TJ ET 0.267 0.267 0.267 rg 150.716 420.114 86.691 0.750 re f 150.716 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 410.259 Td /F1 9.8 Tf [(17.46#)] TJ ET 0.267 0.267 0.267 rg 236.657 420.114 65.124 0.750 re f 236.657 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 410.259 Td /F1 9.8 Tf [(3.67*)] TJ ET 0.267 0.267 0.267 rg 301.032 420.114 72.390 0.750 re f 301.032 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 410.259 Td /F1 9.8 Tf [(149.36#)] TJ ET 0.267 0.267 0.267 rg 372.672 420.114 65.124 0.750 re f 372.672 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 410.259 Td /F1 9.8 Tf [(5.26**)] TJ ET 0.267 0.267 0.267 rg 437.046 420.114 72.390 0.750 re f 437.046 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 410.259 Td /F1 9.8 Tf [(140.32#)] TJ ET 0.267 0.267 0.267 rg 508.686 420.114 72.189 0.750 re f 508.686 404.233 0.750 16.631 re f 580.125 404.233 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 410.259 Td /F1 9.8 Tf [(4.14**)] TJ ET 0.267 0.267 0.267 rg 26.625 404.233 124.841 0.750 re f 26.625 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 394.378 Td /F4 9.8 Tf [(Repeats)] TJ ET 0.267 0.267 0.267 rg 150.716 404.233 86.691 0.750 re f 150.716 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 394.378 Td /F1 9.8 Tf [(83.97#)] TJ ET 0.267 0.267 0.267 rg 236.657 404.233 65.124 0.750 re f 236.657 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 301.032 404.233 72.390 0.750 re f 301.032 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 372.672 404.233 65.124 0.750 re f 372.672 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 404.233 72.390 0.750 re f 437.046 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 394.378 Td /F1 9.8 Tf [(21.80#)] TJ ET 0.267 0.267 0.267 rg 508.686 404.233 72.189 0.750 re f 508.686 388.351 0.750 16.631 re f 580.125 388.351 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 394.378 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 388.351 124.841 0.750 re f 26.625 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 378.496 Td /F4 9.8 Tf [(Alternations)] TJ ET 0.267 0.267 0.267 rg 150.716 388.351 86.691 0.750 re f 150.716 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 378.496 Td /F1 9.8 Tf [(33.25#)] TJ ET 0.267 0.267 0.267 rg 236.657 388.351 65.124 0.750 re f 236.657 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 378.496 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 301.032 388.351 72.390 0.750 re f 301.032 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 378.496 Td /F1 9.8 Tf [(5.66*)] TJ ET 0.267 0.267 0.267 rg 372.672 388.351 65.124 0.750 re f 372.672 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 378.496 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 388.351 72.390 0.750 re f 437.046 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 378.496 Td /F1 9.8 Tf [(7.30*)] TJ ET 0.267 0.267 0.267 rg 508.686 388.351 72.189 0.750 re f 508.686 372.470 0.750 16.631 re f 580.125 372.470 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 378.496 Td /F1 9.8 Tf [(2.39*)] TJ ET 0.267 0.267 0.267 rg 26.625 372.470 124.841 0.750 re f 26.625 356.589 124.841 0.750 re f 26.625 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 362.615 Td /F4 9.8 Tf [(Collected Rewards)] TJ ET 0.267 0.267 0.267 rg 150.716 372.470 86.691 0.750 re f 150.716 356.589 86.691 0.750 re f 150.716 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 155.966 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 236.657 372.470 65.124 0.750 re f 236.657 356.589 65.124 0.750 re f 236.657 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 241.907 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 301.032 372.470 72.390 0.750 re f 301.032 356.589 72.390 0.750 re f 301.032 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.282 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 372.672 372.470 65.124 0.750 re f 372.672 356.589 65.124 0.750 re f 372.672 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 377.922 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 437.046 372.470 72.390 0.750 re f 437.046 356.589 72.390 0.750 re f 437.046 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 442.296 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 508.686 372.470 72.189 0.750 re f 508.686 356.589 72.189 0.750 re f 508.686 356.589 0.750 16.631 re f 580.125 356.589 0.750 16.631 re f 0.271 0.267 0.267 rg BT 513.936 362.615 Td /F1 9.8 Tf [(NS)] TJ ET 0.965 0.965 0.965 rg 26.250 -154.536 555.000 465.750 re f 0.267 0.267 0.267 rg 26.250 311.214 m 581.250 311.214 l 581.250 310.464 l 26.250 310.464 l f q 321.000 0 0 450.000 35.250 -148.536 cm /I7 Do Q q 35.250 -154.536 537.000 0.000 re W n Q Q q 321.000 0 0 450.000 35.250 -148.536 cm /I7 Do Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(6)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Huntington Disease)] TJ ET Q endstream endobj 335 0 obj << /Type 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E&is@+9$Do"x7k7я@Q=/O~?%@1z_$"*8:ɢKc }Շ( ?!y3M~#+O+|#v_ů)/ipn??S2xoǬ?/S2xoQ2o__'v_=+ =&MK5%w?@ӽt 1E=+g ]+ y$~J~'ͧA_zxAR=߉iPWޟD!^=,4.k9$K~%vfσNPO;GD?o3|V1GM2C@ԝ)A@ Hh8I_Z_:? إmwuo@!V+S K'E:/"?oJa%#҄z~ko7_9+w!Im_[)_ u9"oZJ ^,ũ? %-# qI#޸߃-Ǡ(_ Mh{C_Q=/O~%@> }jA5Cǟ$x) ;qOoirWbWIqOoj?_ Tҿ*O4v4zÙ<''1k qm?&f(`m?0%0x&=+/3> [xkL}.?i!y_bөF>Q&'lRXּ=y,v^z=^;UX|eY%^Iɯo؎a."V0y_L?rҸkG7hZl/)~8DžtGZׯ63NܠpO^ا㭗48aGshpzP?i?r~:? xJK%dOqs\mC\t^K|GMh?ࣿdJ_W|a/q^|L xcnkڍ1c0yX\D *׵~@|/8h)|0k<=F+3!ڎeK-F`B eAǻ(jEP?g ]+M8c~`Wr; #V^[j:~gis ˕!Ee8 4OO"% Ian/ؗwڟǞ./_S۽؞āmaF] 2v?s<|&v ^:!moȍ7f2|FF tR\pԃ7ftAe-1̷.S9 zc?Y!?(J%W/j>f-1 A:P#,Gk|Sb/nѝdd I[cw!Im_[)_ uN<$FIo,`[kf62#onx>Wc{@wv;];NHbRIxcN7-IW~U-f>6[uOiw:Cq4b@`BǢڿS,cYT/͉ЀYE% ҸσMtO1몼[FP3Q3RKxJIs.Ǩho !'Wi"#,1߶QC^{j|Z^+ĂTƹHv W'q^8jZ|HSpiڀ> }jA5IujYO-͔Ģd`#CTIRxY,zPqOsirWbWN-r>j,O9___| Z]iOl-/bϓ$VѤHpFAtR+dfׄ|iHJ9~/4K-~fZZAfq*1Ijj#8XPE5qBWw{=Op~?hm A4[yⶹh<-خH2ԏG|AIu]Udž-L2ҀF(qiuI4ǹH!Tpda+sJtX&5)&w 3Aii?5VOTiΝ2Y"2*6kҚ56GPaE`|;X6lb $_2Eٍ{eG$h{ ^(#@Ţ(EQEQEQEQE&Fڊ(M6 (b(ERb(KERQE-(PIE0(Fh M\QEwJ^Q@ KEf(4Q@Q@ endstream endobj 338 0 obj << /Type /Annot /Subtype /Link /A 339 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 -148.5360 356.2500 301.4640 ] >> endobj 339 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Balci_fig5.jpg) >> endobj 340 0 obj << /Type /Annot /Subtype /Link /A 341 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 -148.5360 356.2500 301.4640 ] >> endobj 341 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Balci_fig5.jpg) >> endobj 342 0 obj << /Type /Page /Parent 3 0 R /Annots [ 344 0 R 346 0 R 348 0 R 351 0 R 353 0 R 355 0 R 357 0 R 359 0 R 361 0 R ] /Contents 343 0 R >> endobj 343 0 obj << /Length 34800 >> stream q 15.000 -85.568 577.500 862.568 re W n 0.965 0.965 0.965 rg 26.250 699.136 555.000 77.864 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 699.136 m 581.250 699.136 l 581.250 699.886 l 26.250 699.886 l f q 35.250 710.386 537.000 66.614 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 5: R6/2 behavioral measures as a function of genotype, age, and light cycle.)] TJ ET BT 35.250 748.106 Td /F1 9.8 Tf [(*: significant genotype differences within each light phase, at each age; **: significant genotype effect for each age, )] TJ ET BT 35.250 734.370 Td /F1 9.8 Tf [(independent of cycle; #: significant differences due to light cycle for each genotype and age; ##: significant differences due )] TJ ET BT 35.250 720.634 Td /F1 9.8 Tf [(to light cycle for each age independently of genotype.)] TJ ET Q q 26.250 677.900 555.000 13.736 re W n 0.271 0.267 0.267 rg BT 26.250 680.647 Td /F1 9.8 Tf [(Table 2. Summary of statistical results for the computer vision measures for Experiment 1 \(R6/2\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 646.993 555.000 23.407 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 670.400 m 581.250 670.400 l 580.500 669.650 l 27.000 669.650 l f 581.250 670.400 m 581.250 646.993 l 580.500 646.993 l 580.500 669.650 l f 26.250 670.400 m 26.250 646.993 l 27.000 646.993 l 27.000 669.650 l f 0.271 0.267 0.267 rg BT 33.000 656.324 Td /F1 9.0 Tf [(Notes: *: p< 0.05, **: p< 0.01, #: p< 0.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 548.730 555.000 98.263 re f 0.965 0.965 0.965 rg 27.000 597.124 148.925 49.119 re f 0.267 0.267 0.267 rg 26.625 645.868 149.300 0.750 re f 26.625 633.963 0.750 12.655 re f 26.625 621.683 0.750 12.280 re f 26.625 609.403 0.750 12.280 re f 26.625 596.749 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 636.719 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 175.925 633.963 404.575 12.280 re f 0.267 0.267 0.267 rg 175.925 645.868 88.836 0.750 re f 264.761 645.868 57.620 0.750 re f 322.381 645.868 74.053 0.750 re f 396.434 645.868 57.620 0.750 re f 454.054 645.868 57.620 0.750 re f 511.673 645.868 69.202 0.750 re f 580.125 633.588 0.750 13.030 re f 0.271 0.267 0.267 rg BT 180.425 636.719 Td /F4 9.8 Tf [(R6/2)] TJ ET 0.965 0.965 0.965 rg 175.925 621.683 88.836 12.280 re f 0.267 0.267 0.267 rg 175.925 633.588 88.836 0.750 re f 0.271 0.267 0.267 rg BT 180.425 624.439 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 264.761 621.683 57.620 12.280 re f 0.267 0.267 0.267 rg 264.761 633.588 57.620 0.750 re f 0.271 0.267 0.267 rg BT 269.261 624.439 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 322.381 621.683 74.053 12.280 re f 0.267 0.267 0.267 rg 322.381 633.588 74.053 0.750 re f 0.271 0.267 0.267 rg BT 326.881 624.439 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 396.434 621.683 57.620 12.280 re f 0.267 0.267 0.267 rg 396.434 633.588 57.620 0.750 re f 0.271 0.267 0.267 rg BT 400.934 624.439 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 454.054 621.683 57.620 12.280 re f 0.267 0.267 0.267 rg 454.054 633.588 57.620 0.750 re f 0.271 0.267 0.267 rg BT 458.554 624.439 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 511.673 621.683 68.827 12.280 re f 0.267 0.267 0.267 rg 511.673 633.588 69.202 0.750 re f 580.125 621.308 0.750 13.030 re f 0.271 0.267 0.267 rg BT 516.173 624.439 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 175.925 609.403 88.836 12.280 re f 0.267 0.267 0.267 rg 175.925 621.308 88.836 0.750 re f 0.271 0.267 0.267 rg BT 180.425 612.159 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 264.761 609.403 57.620 12.280 re f 0.267 0.267 0.267 rg 264.761 621.308 57.620 0.750 re f 0.271 0.267 0.267 rg BT 269.261 612.159 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 322.381 609.403 74.053 12.280 re f 0.267 0.267 0.267 rg 322.381 621.308 74.053 0.750 re f 0.271 0.267 0.267 rg BT 326.881 612.159 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 396.434 609.403 57.620 12.280 re f 0.267 0.267 0.267 rg 396.434 621.308 57.620 0.750 re f 0.965 0.965 0.965 rg 454.054 609.403 57.620 12.280 re f 0.267 0.267 0.267 rg 454.054 621.308 57.620 0.750 re f 0.965 0.965 0.965 rg 511.673 609.403 68.827 12.280 re f 0.267 0.267 0.267 rg 511.673 621.308 69.202 0.750 re f 580.125 609.028 0.750 13.030 re f 0.965 0.965 0.965 rg 175.925 597.124 88.836 12.280 re f 0.267 0.267 0.267 rg 175.925 609.028 88.836 0.750 re f 0.271 0.267 0.267 rg BT 180.425 599.880 Td /F4 9.8 Tf [(F\(1,2\))] TJ ET 0.965 0.965 0.965 rg 264.761 597.124 57.620 12.280 re f 0.267 0.267 0.267 rg 264.761 609.028 57.620 0.750 re f 0.271 0.267 0.267 rg BT 269.261 599.880 Td /F4 9.8 Tf [(F\(3,6\))] TJ ET 0.965 0.965 0.965 rg 322.381 597.124 74.053 12.280 re f 0.267 0.267 0.267 rg 322.381 609.028 74.053 0.750 re f 0.271 0.267 0.267 rg BT 326.881 599.880 Td /F4 9.8 Tf [(F\(1,2\))] TJ ET 0.965 0.965 0.965 rg 396.434 597.124 57.620 12.280 re f 0.267 0.267 0.267 rg 396.434 609.028 57.620 0.750 re f 0.271 0.267 0.267 rg BT 400.934 599.880 Td /F4 9.8 Tf [(F\(3,6\))] TJ ET 0.965 0.965 0.965 rg 454.054 597.124 57.620 12.280 re f 0.267 0.267 0.267 rg 454.054 609.028 57.620 0.750 re f 0.271 0.267 0.267 rg BT 458.554 599.880 Td /F4 9.8 Tf [(F\(1,2\))] TJ ET 0.965 0.965 0.965 rg 511.673 597.124 68.827 12.280 re f 0.267 0.267 0.267 rg 511.673 609.028 69.202 0.750 re f 580.125 596.749 0.750 13.030 re f 0.271 0.267 0.267 rg BT 516.173 599.880 Td /F4 9.8 Tf [(F\(6,6\))] TJ ET 0.267 0.267 0.267 rg 26.625 596.749 149.675 0.750 re f 26.625 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 586.894 Td /F4 9.8 Tf [(Locomotion)] TJ ET 0.267 0.267 0.267 rg 175.550 596.749 89.586 0.750 re f 175.550 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 180.800 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 264.386 596.749 58.370 0.750 re f 264.386 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 269.636 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 322.006 596.749 74.803 0.750 re f 322.006 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 327.256 586.894 Td /F1 9.8 Tf [(136.16#)] TJ ET 0.267 0.267 0.267 rg 396.059 596.749 58.370 0.750 re f 396.059 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 401.309 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 453.679 596.749 58.370 0.750 re f 453.679 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 458.929 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 511.298 596.749 69.577 0.750 re f 511.298 580.867 0.750 16.631 re f 580.125 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 516.548 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 580.867 149.675 0.750 re f 26.625 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 571.012 Td /F4 9.8 Tf [(Time Immobile)] TJ ET 0.267 0.267 0.267 rg 175.550 580.867 89.586 0.750 re f 175.550 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 180.800 571.012 Td /F1 9.8 Tf [(205.80#)] TJ ET 0.267 0.267 0.267 rg 264.386 580.867 58.370 0.750 re f 264.386 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 269.636 571.012 Td /F1 9.8 Tf [(6.91*)] TJ ET 0.267 0.267 0.267 rg 322.006 580.867 74.803 0.750 re f 322.006 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 327.256 571.012 Td /F1 9.8 Tf [(28.4*)] TJ ET 0.267 0.267 0.267 rg 396.059 580.867 58.370 0.750 re f 396.059 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 401.309 571.012 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 453.679 580.867 58.370 0.750 re f 453.679 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 458.929 571.012 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 511.298 580.867 69.577 0.750 re f 511.298 564.986 0.750 16.631 re f 580.125 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 516.548 571.012 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 564.986 149.675 0.750 re f 26.625 549.105 149.675 0.750 re f 26.625 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 555.131 Td /F4 9.8 Tf [(Rearing/Climbing)] TJ ET 0.267 0.267 0.267 rg 175.550 564.986 89.586 0.750 re f 175.550 549.105 89.586 0.750 re f 175.550 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 180.800 555.131 Td /F1 9.8 Tf [(45.53*)] TJ ET 0.267 0.267 0.267 rg 264.386 564.986 58.370 0.750 re f 264.386 549.105 58.370 0.750 re f 264.386 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 269.636 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 322.006 564.986 74.803 0.750 re f 322.006 549.105 74.803 0.750 re f 322.006 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 327.256 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 396.059 564.986 58.370 0.750 re f 396.059 549.105 58.370 0.750 re f 396.059 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 401.309 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 453.679 564.986 58.370 0.750 re f 453.679 549.105 58.370 0.750 re f 453.679 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 458.929 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 511.298 564.986 69.577 0.750 re f 511.298 549.105 69.577 0.750 re f 511.298 549.105 0.750 16.631 re f 580.125 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 516.548 555.131 Td /F1 9.8 Tf [(NS)] TJ ET BT 26.250 494.206 Td /F1 9.8 Tf [(Consistent with earlier findings )] TJ ET 0.267 0.267 0.267 rg BT 161.171 495.713 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 165.989 498.094 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 168.399 495.713 Td /F4 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 178.036 494.206 Td /F1 9.8 Tf [(, these computer vision-based measures overall suggested lower locomotor activity and )] TJ ET BT 26.250 482.301 Td /F1 9.8 Tf [(decreased motor function of R6/2 mice compared to WT mice. Details of these results are summarized below and in Table 2.)] TJ ET BT 26.250 462.897 Td /F5 9.8 Tf [(Locomotion)] TJ ET BT 76.648 462.897 Td /F1 9.8 Tf [(. This group measure, obtained from the computer vision system, showed an overall increase in the activity of all )] TJ ET BT 26.250 450.992 Td /F1 9.8 Tf [(mice during the night independently of genotype \(Figure 6A; Table 2, Cycle main effect: )] TJ ET BT 405.603 450.992 Td /F5 9.8 Tf [(p)] TJ ET BT 411.024 450.992 Td /F1 9.8 Tf [( < 0.001\). R6/2 mice showed a slight )] TJ ET BT 26.250 439.087 Td /F1 9.8 Tf [(but not significant decrease in locomotion.)] TJ ET BT 26.250 419.682 Td /F5 9.8 Tf [(Time Immobile)] TJ ET BT 90.171 419.682 Td /F1 9.8 Tf [(. From the computer vision it was also clear that R6/2 mice were more immobile over the whole testing period. As )] TJ ET BT 26.250 407.778 Td /F1 9.8 Tf [(expected both groups were less immobile during the dark phase, while all animals became slightly more immobile with repeated )] TJ ET BT 26.250 395.873 Td /F1 9.8 Tf [(testing \(Figure 6B; Table 2, Genotype, Age and Cycle main effects: )] TJ ET BT 318.350 395.873 Td /F5 9.8 Tf [(p)] TJ ET BT 323.771 395.873 Td /F1 9.8 Tf [( < 0.001\).)] TJ ET BT 26.250 376.468 Td /F5 9.8 Tf [(Rearing/Climbing)] TJ ET BT 101.013 376.468 Td /F1 9.8 Tf [(. R6/2 mice reared and climbed less \(Figure 6C; Table 2, Genotype main effect: )] TJ ET BT 446.738 376.468 Td /F5 9.8 Tf [(p)] TJ ET BT 452.159 376.468 Td /F1 9.8 Tf [( < 0.05\). Although rearing )] TJ ET BT 26.250 364.563 Td /F1 9.8 Tf [(seemed higher during the night there were no significant effects of either cycle or age.)] TJ ET 0.965 0.965 0.965 rg 26.250 -85.568 555.000 440.250 re f 0.267 0.267 0.267 rg 26.250 354.682 m 581.250 354.682 l 581.250 353.932 l 26.250 353.932 l f q 450.000 0 0 424.500 35.250 -79.568 cm /I8 Do Q q 35.250 -85.568 537.000 0.000 re W n Q Q q 15.000 -85.568 577.500 862.568 re W n 0.965 0.965 0.965 rg 26.250 699.136 555.000 77.864 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 699.136 m 581.250 699.136 l 581.250 699.886 l 26.250 699.886 l f q 35.250 710.386 537.000 66.614 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 5: R6/2 behavioral measures as a function of genotype, age, and light cycle.)] TJ ET BT 35.250 748.106 Td /F1 9.8 Tf [(*: significant genotype differences within each light phase, at each age; **: significant genotype effect for each age, )] TJ ET BT 35.250 734.370 Td /F1 9.8 Tf [(independent of cycle; #: significant differences due to light cycle for each genotype and age; ##: significant differences due )] TJ ET BT 35.250 720.634 Td /F1 9.8 Tf [(to light cycle for each age independently of genotype.)] TJ ET Q q 26.250 677.900 555.000 13.736 re W n 0.271 0.267 0.267 rg BT 26.250 680.647 Td /F1 9.8 Tf [(Table 2. Summary of statistical results for the computer vision measures for Experiment 1 \(R6/2\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 646.993 555.000 23.407 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 670.400 m 581.250 670.400 l 580.500 669.650 l 27.000 669.650 l f 581.250 670.400 m 581.250 646.993 l 580.500 646.993 l 580.500 669.650 l f 26.250 670.400 m 26.250 646.993 l 27.000 646.993 l 27.000 669.650 l f 0.271 0.267 0.267 rg BT 33.000 656.324 Td /F1 9.0 Tf [(Notes: *: p< 0.05, **: p< 0.01, #: p< 0.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 548.730 555.000 98.263 re f 0.965 0.965 0.965 rg 27.000 597.124 148.925 49.119 re f 0.267 0.267 0.267 rg 26.625 645.868 149.300 0.750 re f 26.625 633.963 0.750 12.655 re f 26.625 621.683 0.750 12.280 re f 26.625 609.403 0.750 12.280 re f 26.625 596.749 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 636.719 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 175.925 633.963 404.575 12.280 re f 0.267 0.267 0.267 rg 175.925 645.868 88.836 0.750 re f 264.761 645.868 57.620 0.750 re f 322.381 645.868 74.053 0.750 re f 396.434 645.868 57.620 0.750 re f 454.054 645.868 57.620 0.750 re f 511.673 645.868 69.202 0.750 re f 580.125 633.588 0.750 13.030 re f 0.271 0.267 0.267 rg BT 180.425 636.719 Td /F4 9.8 Tf [(R6/2)] TJ ET 0.965 0.965 0.965 rg 175.925 621.683 88.836 12.280 re f 0.267 0.267 0.267 rg 175.925 633.588 88.836 0.750 re f 0.271 0.267 0.267 rg BT 180.425 624.439 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 264.761 621.683 57.620 12.280 re f 0.267 0.267 0.267 rg 264.761 633.588 57.620 0.750 re f 0.271 0.267 0.267 rg BT 269.261 624.439 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 322.381 621.683 74.053 12.280 re f 0.267 0.267 0.267 rg 322.381 633.588 74.053 0.750 re f 0.271 0.267 0.267 rg BT 326.881 624.439 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 396.434 621.683 57.620 12.280 re f 0.267 0.267 0.267 rg 396.434 633.588 57.620 0.750 re f 0.271 0.267 0.267 rg BT 400.934 624.439 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 454.054 621.683 57.620 12.280 re f 0.267 0.267 0.267 rg 454.054 633.588 57.620 0.750 re f 0.271 0.267 0.267 rg BT 458.554 624.439 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 511.673 621.683 68.827 12.280 re f 0.267 0.267 0.267 rg 511.673 633.588 69.202 0.750 re f 580.125 621.308 0.750 13.030 re f 0.271 0.267 0.267 rg BT 516.173 624.439 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 175.925 609.403 88.836 12.280 re f 0.267 0.267 0.267 rg 175.925 621.308 88.836 0.750 re f 0.271 0.267 0.267 rg BT 180.425 612.159 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 264.761 609.403 57.620 12.280 re f 0.267 0.267 0.267 rg 264.761 621.308 57.620 0.750 re f 0.271 0.267 0.267 rg BT 269.261 612.159 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 322.381 609.403 74.053 12.280 re f 0.267 0.267 0.267 rg 322.381 621.308 74.053 0.750 re f 0.271 0.267 0.267 rg BT 326.881 612.159 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 396.434 609.403 57.620 12.280 re f 0.267 0.267 0.267 rg 396.434 621.308 57.620 0.750 re f 0.965 0.965 0.965 rg 454.054 609.403 57.620 12.280 re f 0.267 0.267 0.267 rg 454.054 621.308 57.620 0.750 re f 0.965 0.965 0.965 rg 511.673 609.403 68.827 12.280 re f 0.267 0.267 0.267 rg 511.673 621.308 69.202 0.750 re f 580.125 609.028 0.750 13.030 re f 0.965 0.965 0.965 rg 175.925 597.124 88.836 12.280 re f 0.267 0.267 0.267 rg 175.925 609.028 88.836 0.750 re f 0.271 0.267 0.267 rg BT 180.425 599.880 Td /F4 9.8 Tf [(F\(1,2\))] TJ ET 0.965 0.965 0.965 rg 264.761 597.124 57.620 12.280 re f 0.267 0.267 0.267 rg 264.761 609.028 57.620 0.750 re f 0.271 0.267 0.267 rg BT 269.261 599.880 Td /F4 9.8 Tf [(F\(3,6\))] TJ ET 0.965 0.965 0.965 rg 322.381 597.124 74.053 12.280 re f 0.267 0.267 0.267 rg 322.381 609.028 74.053 0.750 re f 0.271 0.267 0.267 rg BT 326.881 599.880 Td /F4 9.8 Tf [(F\(1,2\))] TJ ET 0.965 0.965 0.965 rg 396.434 597.124 57.620 12.280 re f 0.267 0.267 0.267 rg 396.434 609.028 57.620 0.750 re f 0.271 0.267 0.267 rg BT 400.934 599.880 Td /F4 9.8 Tf [(F\(3,6\))] TJ ET 0.965 0.965 0.965 rg 454.054 597.124 57.620 12.280 re f 0.267 0.267 0.267 rg 454.054 609.028 57.620 0.750 re f 0.271 0.267 0.267 rg BT 458.554 599.880 Td /F4 9.8 Tf [(F\(1,2\))] TJ ET 0.965 0.965 0.965 rg 511.673 597.124 68.827 12.280 re f 0.267 0.267 0.267 rg 511.673 609.028 69.202 0.750 re f 580.125 596.749 0.750 13.030 re f 0.271 0.267 0.267 rg BT 516.173 599.880 Td /F4 9.8 Tf [(F\(6,6\))] TJ ET 0.267 0.267 0.267 rg 26.625 596.749 149.675 0.750 re f 26.625 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 586.894 Td /F4 9.8 Tf [(Locomotion)] TJ ET 0.267 0.267 0.267 rg 175.550 596.749 89.586 0.750 re f 175.550 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 180.800 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 264.386 596.749 58.370 0.750 re f 264.386 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 269.636 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 322.006 596.749 74.803 0.750 re f 322.006 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 327.256 586.894 Td /F1 9.8 Tf [(136.16#)] TJ ET 0.267 0.267 0.267 rg 396.059 596.749 58.370 0.750 re f 396.059 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 401.309 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 453.679 596.749 58.370 0.750 re f 453.679 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 458.929 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 511.298 596.749 69.577 0.750 re f 511.298 580.867 0.750 16.631 re f 580.125 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 516.548 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 580.867 149.675 0.750 re f 26.625 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 571.012 Td /F4 9.8 Tf [(Time Immobile)] TJ ET 0.267 0.267 0.267 rg 175.550 580.867 89.586 0.750 re f 175.550 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 180.800 571.012 Td /F1 9.8 Tf [(205.80#)] TJ ET 0.267 0.267 0.267 rg 264.386 580.867 58.370 0.750 re f 264.386 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 269.636 571.012 Td /F1 9.8 Tf [(6.91*)] TJ ET 0.267 0.267 0.267 rg 322.006 580.867 74.803 0.750 re f 322.006 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 327.256 571.012 Td /F1 9.8 Tf [(28.4*)] TJ ET 0.267 0.267 0.267 rg 396.059 580.867 58.370 0.750 re f 396.059 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 401.309 571.012 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 453.679 580.867 58.370 0.750 re f 453.679 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 458.929 571.012 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 511.298 580.867 69.577 0.750 re f 511.298 564.986 0.750 16.631 re f 580.125 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 516.548 571.012 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 564.986 149.675 0.750 re f 26.625 549.105 149.675 0.750 re f 26.625 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 555.131 Td /F4 9.8 Tf [(Rearing/Climbing)] TJ ET 0.267 0.267 0.267 rg 175.550 564.986 89.586 0.750 re f 175.550 549.105 89.586 0.750 re f 175.550 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 180.800 555.131 Td /F1 9.8 Tf [(45.53*)] TJ ET 0.267 0.267 0.267 rg 264.386 564.986 58.370 0.750 re f 264.386 549.105 58.370 0.750 re f 264.386 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 269.636 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 322.006 564.986 74.803 0.750 re f 322.006 549.105 74.803 0.750 re f 322.006 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 327.256 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 396.059 564.986 58.370 0.750 re f 396.059 549.105 58.370 0.750 re f 396.059 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 401.309 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 453.679 564.986 58.370 0.750 re f 453.679 549.105 58.370 0.750 re f 453.679 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 458.929 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 511.298 564.986 69.577 0.750 re f 511.298 549.105 69.577 0.750 re f 511.298 549.105 0.750 16.631 re f 580.125 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 516.548 555.131 Td /F1 9.8 Tf [(NS)] TJ ET BT 26.250 494.206 Td /F1 9.8 Tf [(Consistent with earlier findings )] TJ ET 0.267 0.267 0.267 rg BT 161.171 495.713 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 165.989 498.094 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 168.399 495.713 Td /F4 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 178.036 494.206 Td /F1 9.8 Tf [(, these computer vision-based measures overall suggested lower locomotor activity and )] TJ ET BT 26.250 482.301 Td /F1 9.8 Tf [(decreased motor function of R6/2 mice compared to WT mice. Details of these results are summarized below and in Table 2.)] TJ ET BT 26.250 462.897 Td /F5 9.8 Tf [(Locomotion)] TJ ET BT 76.648 462.897 Td /F1 9.8 Tf [(. This group measure, obtained from the computer vision system, showed an overall increase in the activity of all )] TJ ET BT 26.250 450.992 Td /F1 9.8 Tf [(mice during the night independently of genotype \(Figure 6A; Table 2, Cycle main effect: )] TJ ET BT 405.603 450.992 Td /F5 9.8 Tf [(p)] TJ ET BT 411.024 450.992 Td /F1 9.8 Tf [( < 0.001\). R6/2 mice showed a slight )] TJ ET BT 26.250 439.087 Td /F1 9.8 Tf [(but not significant decrease in locomotion.)] TJ ET BT 26.250 419.682 Td /F5 9.8 Tf [(Time Immobile)] TJ ET BT 90.171 419.682 Td /F1 9.8 Tf [(. From the computer vision it was also clear that R6/2 mice were more immobile over the whole testing period. As )] TJ ET BT 26.250 407.778 Td /F1 9.8 Tf [(expected both groups were less immobile during the dark phase, while all animals became slightly more immobile with repeated )] TJ ET BT 26.250 395.873 Td /F1 9.8 Tf [(testing \(Figure 6B; Table 2, Genotype, Age and Cycle main effects: )] TJ ET BT 318.350 395.873 Td /F5 9.8 Tf [(p)] TJ ET BT 323.771 395.873 Td /F1 9.8 Tf [( < 0.001\).)] TJ ET BT 26.250 376.468 Td /F5 9.8 Tf [(Rearing/Climbing)] TJ ET BT 101.013 376.468 Td /F1 9.8 Tf [(. R6/2 mice reared and climbed less \(Figure 6C; Table 2, Genotype main effect: )] TJ ET BT 446.738 376.468 Td /F5 9.8 Tf [(p)] TJ ET BT 452.159 376.468 Td /F1 9.8 Tf [( < 0.05\). Although rearing )] TJ ET BT 26.250 364.563 Td /F1 9.8 Tf [(seemed higher during the night there were no significant effects of either cycle or age.)] TJ ET 0.965 0.965 0.965 rg 26.250 -85.568 555.000 440.250 re f 0.267 0.267 0.267 rg 26.250 354.682 m 581.250 354.682 l 581.250 353.932 l 26.250 353.932 l f q 450.000 0 0 424.500 35.250 -79.568 cm /I8 Do Q q 35.250 -85.568 537.000 0.000 re W n Q Q q 15.000 -85.568 577.500 862.568 re W n 0.965 0.965 0.965 rg 26.250 699.136 555.000 77.864 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 699.136 m 581.250 699.136 l 581.250 699.886 l 26.250 699.886 l f q 35.250 710.386 537.000 66.614 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 5: R6/2 behavioral measures as a function of genotype, age, and light cycle.)] TJ ET BT 35.250 748.106 Td /F1 9.8 Tf [(*: significant genotype differences within each light phase, at each age; **: significant genotype effect for each age, )] TJ ET BT 35.250 734.370 Td /F1 9.8 Tf [(independent of cycle; #: significant differences due to light cycle for each genotype and age; ##: significant differences due )] TJ ET BT 35.250 720.634 Td /F1 9.8 Tf [(to light cycle for each age independently of genotype.)] TJ ET Q q 26.250 677.900 555.000 13.736 re W n 0.271 0.267 0.267 rg BT 26.250 680.647 Td /F1 9.8 Tf [(Table 2. Summary of statistical results for the computer vision measures for Experiment 1 \(R6/2\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 646.993 555.000 23.407 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 670.400 m 581.250 670.400 l 580.500 669.650 l 27.000 669.650 l f 581.250 670.400 m 581.250 646.993 l 580.500 646.993 l 580.500 669.650 l f 26.250 670.400 m 26.250 646.993 l 27.000 646.993 l 27.000 669.650 l f 0.271 0.267 0.267 rg BT 33.000 656.324 Td /F1 9.0 Tf [(Notes: *: p< 0.05, **: p< 0.01, #: p< 0.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 548.730 555.000 98.263 re f 0.965 0.965 0.965 rg 27.000 597.124 148.925 49.119 re f 0.267 0.267 0.267 rg 26.625 645.868 149.300 0.750 re f 26.625 633.963 0.750 12.655 re f 26.625 621.683 0.750 12.280 re f 26.625 609.403 0.750 12.280 re f 26.625 596.749 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 636.719 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 175.925 633.963 404.575 12.280 re f 0.267 0.267 0.267 rg 175.925 645.868 88.836 0.750 re f 264.761 645.868 57.620 0.750 re f 322.381 645.868 74.053 0.750 re f 396.434 645.868 57.620 0.750 re f 454.054 645.868 57.620 0.750 re f 511.673 645.868 69.202 0.750 re f 580.125 633.588 0.750 13.030 re f 0.271 0.267 0.267 rg BT 180.425 636.719 Td /F4 9.8 Tf [(R6/2)] TJ ET 0.965 0.965 0.965 rg 175.925 621.683 88.836 12.280 re f 0.267 0.267 0.267 rg 175.925 633.588 88.836 0.750 re f 0.271 0.267 0.267 rg BT 180.425 624.439 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 264.761 621.683 57.620 12.280 re f 0.267 0.267 0.267 rg 264.761 633.588 57.620 0.750 re f 0.271 0.267 0.267 rg BT 269.261 624.439 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 322.381 621.683 74.053 12.280 re f 0.267 0.267 0.267 rg 322.381 633.588 74.053 0.750 re f 0.271 0.267 0.267 rg BT 326.881 624.439 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 396.434 621.683 57.620 12.280 re f 0.267 0.267 0.267 rg 396.434 633.588 57.620 0.750 re f 0.271 0.267 0.267 rg BT 400.934 624.439 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 454.054 621.683 57.620 12.280 re f 0.267 0.267 0.267 rg 454.054 633.588 57.620 0.750 re f 0.271 0.267 0.267 rg BT 458.554 624.439 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 511.673 621.683 68.827 12.280 re f 0.267 0.267 0.267 rg 511.673 633.588 69.202 0.750 re f 580.125 621.308 0.750 13.030 re f 0.271 0.267 0.267 rg BT 516.173 624.439 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 175.925 609.403 88.836 12.280 re f 0.267 0.267 0.267 rg 175.925 621.308 88.836 0.750 re f 0.271 0.267 0.267 rg BT 180.425 612.159 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 264.761 609.403 57.620 12.280 re f 0.267 0.267 0.267 rg 264.761 621.308 57.620 0.750 re f 0.271 0.267 0.267 rg BT 269.261 612.159 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 322.381 609.403 74.053 12.280 re f 0.267 0.267 0.267 rg 322.381 621.308 74.053 0.750 re f 0.271 0.267 0.267 rg BT 326.881 612.159 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 396.434 609.403 57.620 12.280 re f 0.267 0.267 0.267 rg 396.434 621.308 57.620 0.750 re f 0.965 0.965 0.965 rg 454.054 609.403 57.620 12.280 re f 0.267 0.267 0.267 rg 454.054 621.308 57.620 0.750 re f 0.965 0.965 0.965 rg 511.673 609.403 68.827 12.280 re f 0.267 0.267 0.267 rg 511.673 621.308 69.202 0.750 re f 580.125 609.028 0.750 13.030 re f 0.965 0.965 0.965 rg 175.925 597.124 88.836 12.280 re f 0.267 0.267 0.267 rg 175.925 609.028 88.836 0.750 re f 0.271 0.267 0.267 rg BT 180.425 599.880 Td /F4 9.8 Tf [(F\(1,2\))] TJ ET 0.965 0.965 0.965 rg 264.761 597.124 57.620 12.280 re f 0.267 0.267 0.267 rg 264.761 609.028 57.620 0.750 re f 0.271 0.267 0.267 rg BT 269.261 599.880 Td /F4 9.8 Tf [(F\(3,6\))] TJ ET 0.965 0.965 0.965 rg 322.381 597.124 74.053 12.280 re f 0.267 0.267 0.267 rg 322.381 609.028 74.053 0.750 re f 0.271 0.267 0.267 rg BT 326.881 599.880 Td /F4 9.8 Tf [(F\(1,2\))] TJ ET 0.965 0.965 0.965 rg 396.434 597.124 57.620 12.280 re f 0.267 0.267 0.267 rg 396.434 609.028 57.620 0.750 re f 0.271 0.267 0.267 rg BT 400.934 599.880 Td /F4 9.8 Tf [(F\(3,6\))] TJ ET 0.965 0.965 0.965 rg 454.054 597.124 57.620 12.280 re f 0.267 0.267 0.267 rg 454.054 609.028 57.620 0.750 re f 0.271 0.267 0.267 rg BT 458.554 599.880 Td /F4 9.8 Tf [(F\(1,2\))] TJ ET 0.965 0.965 0.965 rg 511.673 597.124 68.827 12.280 re f 0.267 0.267 0.267 rg 511.673 609.028 69.202 0.750 re f 580.125 596.749 0.750 13.030 re f 0.271 0.267 0.267 rg BT 516.173 599.880 Td /F4 9.8 Tf [(F\(6,6\))] TJ ET 0.267 0.267 0.267 rg 26.625 596.749 149.675 0.750 re f 26.625 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 586.894 Td /F4 9.8 Tf [(Locomotion)] TJ ET 0.267 0.267 0.267 rg 175.550 596.749 89.586 0.750 re f 175.550 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 180.800 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 264.386 596.749 58.370 0.750 re f 264.386 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 269.636 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 322.006 596.749 74.803 0.750 re f 322.006 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 327.256 586.894 Td /F1 9.8 Tf [(136.16#)] TJ ET 0.267 0.267 0.267 rg 396.059 596.749 58.370 0.750 re f 396.059 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 401.309 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 453.679 596.749 58.370 0.750 re f 453.679 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 458.929 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 511.298 596.749 69.577 0.750 re f 511.298 580.867 0.750 16.631 re f 580.125 580.867 0.750 16.631 re f 0.271 0.267 0.267 rg BT 516.548 586.894 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 580.867 149.675 0.750 re f 26.625 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 571.012 Td /F4 9.8 Tf [(Time Immobile)] TJ ET 0.267 0.267 0.267 rg 175.550 580.867 89.586 0.750 re f 175.550 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 180.800 571.012 Td /F1 9.8 Tf [(205.80#)] TJ ET 0.267 0.267 0.267 rg 264.386 580.867 58.370 0.750 re f 264.386 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 269.636 571.012 Td /F1 9.8 Tf [(6.91*)] TJ ET 0.267 0.267 0.267 rg 322.006 580.867 74.803 0.750 re f 322.006 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 327.256 571.012 Td /F1 9.8 Tf [(28.4*)] TJ ET 0.267 0.267 0.267 rg 396.059 580.867 58.370 0.750 re f 396.059 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 401.309 571.012 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 453.679 580.867 58.370 0.750 re f 453.679 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 458.929 571.012 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 511.298 580.867 69.577 0.750 re f 511.298 564.986 0.750 16.631 re f 580.125 564.986 0.750 16.631 re f 0.271 0.267 0.267 rg BT 516.548 571.012 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 564.986 149.675 0.750 re f 26.625 549.105 149.675 0.750 re f 26.625 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 555.131 Td /F4 9.8 Tf [(Rearing/Climbing)] TJ ET 0.267 0.267 0.267 rg 175.550 564.986 89.586 0.750 re f 175.550 549.105 89.586 0.750 re f 175.550 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 180.800 555.131 Td /F1 9.8 Tf [(45.53*)] TJ ET 0.267 0.267 0.267 rg 264.386 564.986 58.370 0.750 re f 264.386 549.105 58.370 0.750 re f 264.386 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 269.636 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 322.006 564.986 74.803 0.750 re f 322.006 549.105 74.803 0.750 re f 322.006 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 327.256 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 396.059 564.986 58.370 0.750 re f 396.059 549.105 58.370 0.750 re f 396.059 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 401.309 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 453.679 564.986 58.370 0.750 re f 453.679 549.105 58.370 0.750 re f 453.679 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 458.929 555.131 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 511.298 564.986 69.577 0.750 re f 511.298 549.105 69.577 0.750 re f 511.298 549.105 0.750 16.631 re f 580.125 549.105 0.750 16.631 re f 0.271 0.267 0.267 rg BT 516.548 555.131 Td /F1 9.8 Tf [(NS)] TJ ET BT 26.250 494.206 Td /F1 9.8 Tf [(Consistent with earlier findings )] TJ ET 0.267 0.267 0.267 rg BT 161.171 495.713 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 165.989 498.094 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 168.399 495.713 Td /F4 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 178.036 494.206 Td /F1 9.8 Tf [(, these computer vision-based measures overall suggested lower locomotor activity and )] TJ ET BT 26.250 482.301 Td /F1 9.8 Tf [(decreased motor function of R6/2 mice compared to WT mice. Details of these results are summarized below and in Table 2.)] TJ ET BT 26.250 462.897 Td /F5 9.8 Tf [(Locomotion)] TJ ET BT 76.648 462.897 Td /F1 9.8 Tf [(. This group measure, obtained from the computer vision system, showed an overall increase in the activity of all )] TJ ET BT 26.250 450.992 Td /F1 9.8 Tf [(mice during the night independently of genotype \(Figure 6A; Table 2, Cycle main effect: )] TJ ET BT 405.603 450.992 Td /F5 9.8 Tf [(p)] TJ ET BT 411.024 450.992 Td /F1 9.8 Tf [( < 0.001\). R6/2 mice showed a slight )] TJ ET BT 26.250 439.087 Td /F1 9.8 Tf [(but not significant decrease in locomotion.)] TJ ET BT 26.250 419.682 Td /F5 9.8 Tf [(Time Immobile)] TJ ET BT 90.171 419.682 Td /F1 9.8 Tf [(. From the computer vision it was also clear that R6/2 mice were more immobile over the whole testing period. As )] TJ ET BT 26.250 407.778 Td /F1 9.8 Tf [(expected both groups were less immobile during the dark phase, while all animals became slightly more immobile with repeated )] TJ ET BT 26.250 395.873 Td /F1 9.8 Tf [(testing \(Figure 6B; Table 2, Genotype, Age and Cycle main effects: )] TJ ET BT 318.350 395.873 Td /F5 9.8 Tf [(p)] TJ ET BT 323.771 395.873 Td /F1 9.8 Tf [( < 0.001\).)] TJ ET BT 26.250 376.468 Td /F5 9.8 Tf [(Rearing/Climbing)] TJ ET BT 101.013 376.468 Td /F1 9.8 Tf [(. R6/2 mice reared and climbed less \(Figure 6C; Table 2, Genotype main effect: )] TJ ET BT 446.738 376.468 Td /F5 9.8 Tf [(p)] TJ ET BT 452.159 376.468 Td /F1 9.8 Tf [( < 0.05\). Although rearing )] TJ ET BT 26.250 364.563 Td /F1 9.8 Tf [(seemed higher during the night there were no significant effects of either cycle or age.)] TJ ET 0.965 0.965 0.965 rg 26.250 -85.568 555.000 440.250 re f 0.267 0.267 0.267 rg 26.250 354.682 m 581.250 354.682 l 581.250 353.932 l 26.250 353.932 l f q 450.000 0 0 424.500 35.250 -79.568 cm /I8 Do Q q 35.250 -85.568 537.000 0.000 re W n Q Q q 450.000 0 0 424.500 35.250 -79.568 cm /I8 Do Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(7)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Huntington Disease)] TJ ET Q endstream endobj 344 0 obj << /Type /Annot /Subtype /Link /A 345 0 R /Border [0 0 0] /H /I /Rect [ 161.1705 494.9117 165.9892 503.7300 ] >> endobj 345 0 obj << /Type /Action >> endobj 346 0 obj << /Type /Annot /Subtype /Link /A 347 0 R /Border [0 0 0] /H /I /Rect [ 168.3985 494.9117 178.0358 503.7300 ] >> endobj 347 0 obj << /Type /Action >> endobj 348 0 obj << /Type /Annot /Subtype /Link /A 349 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 -79.5675 485.2500 344.9325 ] >> endobj 349 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Balci_fig6.jpg) >> endobj 350 0 obj << /Type /XObject /Subtype /Image /Width 600 /Height 566 /ColorSpace /DeviceRGB /Filter /DCTDecode /BitsPerComponent 8 /Length 57724>> stream JFIF;CREATOR: gd-jpeg v1.0 (using IJG JPEG v62), quality = 90 C     C   6X" }!1AQa"q2#BR$3br %&'()*456789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz w!1AQaq"2B #3Rbr $4%&'()*56789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz ?S(((((((((((((((((((((((((k:ŗGQ \LR5I'.e*RnK-$Cс(Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@Q@*[G/<9h<{gg W o:&,_L`1]7+C?Tg/9oczWٳ|cL|$h4?K W`S{أsw| _i՟[cHt|Cu6q xO_vߴOWo#Tݍ{; dl~Mo"ڬ?OtUJI`mI f*H |S~5_VxKW:fq1X?3`* |EOڃfdzm5@iMsC?#[{OPdٟhggZw<K⵲v[ڡY bFpOWk? >)yh_>]&|pλ|}o5? f/hGZzHyݥb}p? 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dWnSB\V PQE V\N"nf8A^mvu_^hwEUP6$קb#'Z''_!Q_^ oGnZ\}GUscoAX&?dL'|faƵkt躗/ŸQ7?ƻ+ ¿B0=h( 2l]Ioot^gg+\) J-Q@gjcۄf1Ѧ =E泺)m t8Ы)U^(ӵ8Ӭ$B-$nޤznh(r{Pa W/W֟L/^ǁ18rժ@þ-4W_ kK| Z??h/k=𾍧.!n/8yVpB捱LWiI/Pdr5A(w@ G[ėQBYCߪS-}AȠHTjK zI?$ C\Jеo~eR-2i0MkHQr+ B3FIt|c߂5Z k=8ؾ=OxZ$ +χ~~<;q閑}I)Uv'CR1I_RR)xNලдS>*_JmL`r!z߈V:ֳ_Iih=G+?v`͊AIL6ƾ!G;8@o-a2I!bq'5w)صzYH͔R C`d\V؈/L=-Z/yW(((j7_?gxV;E6*XǦ-/aR/_xcP0c?vҙAqmq*E# KFrf=)<ޢ͢V>[=G◍5C v7l?C7 *׆ VB#s IH Zki7V^ޕ7W1D$Wj`2 +F* ER(Ǿ ]=sPѵ7L]$N'6Z)8)r<~ xdg\x+[[&mXdXmVbRK`~1_xc 'Y4Zk&/k%s4 pIB'j1zSODUm\|YszG_z -|S+YCm mk?$/+J9 G4ҙ/.ۦ sÜu=F ,a@=uGQE ((~&Ayu3i)l &1_|=GԴ',l3Fe4Q\E<r#CiHдwV|=7㍿_\hj,ɣ-e]}eBpq -TD Ԟ#hhʈU|d8uFъKbl-!Ϝ?i~7/5=;6MJkImVt.F.<15xG7Gk;lWFY\)"Ao~}aRjYk}ep9cG" +/|;ewO\]_^5s~sd']&s剭c$wm6i#"nZ_7V)ʘFܬ€޳xM[i|lXbI $zxP$QE ( (6!W>!Zi*^R4mȣr+Z6ҷ_kc kڕ׈䵶tP[&GJC"eQ}GMq6yoxnˤ"|Đ*P8+1ҕ"PWԛ-QEyW%'|-o Z|^"+O#;WOMY/[i7IJnl|a~i.HcSQm}US5Lͫ SIg^(EI]|Ϙ GϋSɼ,;x,e`z.jA_Z]}[DOuQ@Š((((((((((((((((((((((((((((((((((((((( endstream endobj 351 0 obj << /Type /Annot /Subtype /Link /A 352 0 R /Border [0 0 0] /H /I /Rect [ 161.1705 494.9117 165.9892 503.7300 ] >> endobj 352 0 obj << /Type /Action >> endobj 353 0 obj << /Type /Annot /Subtype /Link /A 354 0 R /Border [0 0 0] /H /I /Rect [ 168.3985 494.9117 178.0358 503.7300 ] >> endobj 354 0 obj << /Type /Action >> endobj 355 0 obj << /Type /Annot /Subtype /Link /A 356 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 -79.5675 485.2500 344.9325 ] >> endobj 356 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Balci_fig6.jpg) >> endobj 357 0 obj << /Type /Annot /Subtype /Link /A 358 0 R /Border [0 0 0] /H /I /Rect [ 161.1705 494.9117 165.9892 503.7300 ] >> endobj 358 0 obj << /Type /Action >> endobj 359 0 obj << /Type /Annot /Subtype /Link /A 360 0 R /Border [0 0 0] /H /I /Rect [ 168.3985 494.9117 178.0358 503.7300 ] >> endobj 360 0 obj << /Type /Action >> endobj 361 0 obj << /Type /Annot /Subtype /Link /A 362 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 -79.5675 485.2500 344.9325 ] >> endobj 362 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Balci_fig6.jpg) >> endobj 363 0 obj << /Type /Page /Parent 3 0 R /Annots [ 365 0 R 368 0 R 370 0 R ] /Contents 364 0 R >> endobj 364 0 obj << /Length 50398 >> stream q 15.000 -334.570 577.500 1111.570 re W n 0.965 0.965 0.965 rg 26.250 700.968 555.000 76.032 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 700.968 m 581.250 700.968 l 581.250 701.718 l 26.250 701.718 l f q 35.250 712.218 537.000 64.782 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 6: Behavioral measures derived from the computer vision of Experiment 1 \(R6/2\) as a function of genotype, )] TJ ET BT 35.250 755.571 Td /F4 9.8 Tf [(age, and light cycle.)] TJ ET BT 35.250 736.201 Td /F1 9.8 Tf [(***: significant genotype effect, independent of cycle or age; ##: significant differences due to light cycle for each age )] TJ ET BT 35.250 722.465 Td /F1 9.8 Tf [(independently of genotype.)] TJ ET Q BT 26.250 683.944 Td /F4 9.8 Tf [(Experiment 2. Analysis of BACHD mice:)] TJ ET BT 26.250 664.539 Td /F5 9.8 Tf [(Corner Entries:)] TJ ET BT 91.809 664.539 Td /F1 9.8 Tf [( BACHD mice exhibited a different phenotype than R6/2 mice in terms of the degree of corner visits: R6/2 mice )] TJ ET BT 26.250 652.635 Td /F1 9.8 Tf [(did not differ from their WT controls in terms of the frequency of corner visits however, BACHD mice entered the corners )] TJ ET BT 26.250 640.730 Td /F1 9.8 Tf [(significantly less than WT mice at all ages and cycle phases with the exception of the dark phase at 16 weeks of age \(Figure )] TJ ET BT 26.250 628.825 Td /F1 9.8 Tf [(7A; Table 3; Genotype x Age x Cycle interaction: )] TJ ET BT 239.229 628.825 Td /F5 9.8 Tf [(p)] TJ ET BT 244.650 628.825 Td /F1 9.8 Tf [(< 001; simple main effects: )] TJ ET BT 363.600 628.825 Td /F5 9.8 Tf [(p)] TJ ET BT 369.021 628.825 Td /F1 9.8 Tf [(s < 0.05\). Both groups were more active during )] TJ ET BT 26.250 616.920 Td /F1 9.8 Tf [(the dark period \(Cycle main effect: )] TJ ET BT 177.434 616.920 Td /F5 9.8 Tf [(p)] TJ ET BT 182.855 616.920 Td /F1 9.8 Tf [(< 0.001\).)] TJ ET BT 26.250 597.516 Td /F5 9.8 Tf [(Licking:)] TJ ET BT 59.303 597.516 Td /F1 9.8 Tf [( Similar to the phenotype of R6/2 mice, BACHD mice exhibited increased thirst, which was not detected at the older )] TJ ET BT 26.250 585.611 Td /F1 9.8 Tf [(ages \(i.e., 10 weeks of age for R6/2 mice and 52 weeks of age for BACHD mice\). At earlier ages, the BACHD mice licked more )] TJ ET BT 26.250 573.706 Td /F1 9.8 Tf [(than did WT mice irrespective of cycle phase, but this difference waned as testing progressed \(Figure 7B; Table 3; Genotype x )] TJ ET BT 26.250 561.801 Td /F1 9.8 Tf [(Age x Cycle interaction: )] TJ ET BT 131.375 561.801 Td /F5 9.8 Tf [(p)] TJ ET BT 136.796 561.801 Td /F1 9.8 Tf [( < 0.01; simple main effects: )] TJ ET BT 261.166 561.801 Td /F5 9.8 Tf [(p)] TJ ET BT 266.587 561.801 Td /F1 9.8 Tf [(s < 0.05\). As expected, both groups licked more during the night period )] TJ ET BT 26.250 549.897 Td /F1 9.8 Tf [(\(with the exemption of WT mice at 36 weeks of age\) and, unexpectedly, licked less as testing progressed \(Age main effect: )] TJ ET BT 557.859 549.897 Td /F5 9.8 Tf [(p)] TJ ET BT 563.280 549.897 Td /F1 9.8 Tf [( < )] TJ ET BT 26.250 537.992 Td /F1 9.8 Tf [(0.001\).)] TJ ET BT 26.250 518.587 Td /F5 9.8 Tf [(Repeat visits:)] TJ ET BT 84.769 518.587 Td /F1 9.8 Tf [( Perseverative responding was apparent only during early test ages for BACHD mice. BACHD mice re-entered )] TJ ET BT 26.250 506.682 Td /F1 9.8 Tf [(previously visited corners more frequently than did WT controls at 16 and 24 weeks of age but less at 52 weeks of age \(Figure )] TJ ET BT 26.250 494.778 Td /F1 9.8 Tf [(7C; Table 3; Genotype x Age interaction: )] TJ ET BT 205.094 494.778 Td /F5 9.8 Tf [(p)] TJ ET BT 210.515 494.778 Td /F1 9.8 Tf [( < 0.001; simple main effects: )] TJ ET BT 340.307 494.778 Td /F5 9.8 Tf [(p)] TJ ET BT 345.728 494.778 Td /F1 9.8 Tf [(s < 0.05\). Both groups made more repeats when they )] TJ ET BT 26.250 482.873 Td /F1 9.8 Tf [(were active during the dark phase and made fewer repeats as testing continued \(Age and Cycle main effect: )] TJ ET BT 495.020 482.873 Td /F5 9.8 Tf [(p)] TJ ET BT 500.441 482.873 Td /F1 9.8 Tf [(s < 0.001\).)] TJ ET BT 26.250 463.468 Td /F5 9.8 Tf [(Alternation. )] TJ ET BT 78.276 463.468 Td /F1 9.8 Tf [(Similar age-dependent patterns of genotypic differences were also observed with spatial alternation performance. )] TJ ET BT 26.250 451.563 Td /F1 9.8 Tf [(BACHD mice alternated less than WT during both cycle phases at 16 weeks of age but the effect was not apparent at later )] TJ ET BT 26.250 439.659 Td /F1 9.8 Tf [(ages. BACHD mice alternated more during the light phase at all ages, while this difference was only observed in WT controls at )] TJ ET BT 26.250 427.754 Td /F1 9.8 Tf [(16 weeks of age. \(Figure 7D; Table 3; Genotype x Age x Cycle interaction: )] TJ ET BT 349.774 427.754 Td /F5 9.8 Tf [(p)] TJ ET BT 355.195 427.754 Td /F1 9.8 Tf [( < 0.001\).)] TJ ET BT 26.250 408.349 Td /F5 9.8 Tf [(Rewards collected: )] TJ ET BT 111.319 408.349 Td /F1 9.8 Tf [(Interestingly, BACHD mice exhibited higher efficiency of reward collection than WT animals, a characteristic )] TJ ET BT 26.250 396.444 Td /F1 9.8 Tf [(that became more pronounced with age. For instance, starting at 24 weeks of age BACHD mice were more likely than WT mice )] TJ ET BT 26.250 384.540 Td /F1 9.8 Tf [(to nosepoke in active corners. Both groups, surprisingly, were slightly but consistently less likely to collect rewards during the )] TJ ET BT 26.250 372.635 Td /F1 9.8 Tf [(dark phase, particularly the BACHD mice \(Figure 7E; Table 3; Genotype x Age and Genotype x Cycle interactions: )] TJ ET BT 521.540 372.635 Td /F5 9.8 Tf [(p)] TJ ET BT 526.961 372.635 Td /F1 9.8 Tf [(s < 0.05; )] TJ ET BT 26.250 360.730 Td /F1 9.8 Tf [(simple main effect: )] TJ ET BT 110.237 360.730 Td /F5 9.8 Tf [(p)] TJ ET BT 115.658 360.730 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET q 26.250 337.113 555.000 13.736 re W n 0.271 0.267 0.267 rg BT 26.250 339.860 Td /F1 9.8 Tf [(Table 3. Summary of the statistical analyses for individual behavioral measures for Experiment 2 \(BACHD\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 306.205 555.000 23.408 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 329.613 m 581.250 329.613 l 580.500 328.863 l 27.000 328.863 l f 581.250 329.613 m 581.250 306.205 l 580.500 306.205 l 580.500 328.863 l f 26.250 329.613 m 26.250 306.205 l 27.000 306.205 l 27.000 328.863 l f 0.271 0.267 0.267 rg BT 33.000 315.537 Td /F1 9.0 Tf [(Notes: *: p< 0.05, **: p< 0.01, #: p< 0.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 176.180 555.000 130.025 re f 0.965 0.965 0.965 rg 27.000 256.336 120.160 49.119 re f 0.267 0.267 0.267 rg 26.625 305.080 120.535 0.750 re f 26.625 293.176 0.750 12.655 re f 26.625 280.896 0.750 12.280 re f 26.625 268.616 0.750 12.280 re f 26.625 255.961 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 295.932 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 147.160 293.176 433.340 12.280 re f 0.267 0.267 0.267 rg 147.160 305.080 83.753 0.750 re f 230.913 305.080 71.149 0.750 re f 302.062 305.080 69.816 0.750 re f 371.878 305.080 71.149 0.750 re f 443.026 305.080 62.736 0.750 re f 505.762 305.080 75.113 0.750 re f 580.125 292.801 0.750 13.030 re f 0.271 0.267 0.267 rg BT 151.660 295.932 Td /F4 9.8 Tf [(BACHD)] TJ ET 0.965 0.965 0.965 rg 147.160 280.896 83.753 12.280 re f 0.267 0.267 0.267 rg 147.160 292.801 83.753 0.750 re f 0.271 0.267 0.267 rg BT 151.660 283.652 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 230.913 280.896 71.149 12.280 re f 0.267 0.267 0.267 rg 230.913 292.801 71.149 0.750 re f 0.271 0.267 0.267 rg BT 235.413 283.652 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 302.062 280.896 69.816 12.280 re f 0.267 0.267 0.267 rg 302.062 292.801 69.816 0.750 re f 0.271 0.267 0.267 rg BT 306.562 283.652 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 371.878 280.896 71.149 12.280 re f 0.267 0.267 0.267 rg 371.878 292.801 71.149 0.750 re f 0.271 0.267 0.267 rg BT 376.378 283.652 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 443.026 280.896 62.736 12.280 re f 0.267 0.267 0.267 rg 443.026 292.801 62.736 0.750 re f 0.271 0.267 0.267 rg BT 447.526 283.652 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 505.762 280.896 74.738 12.280 re f 0.267 0.267 0.267 rg 505.762 292.801 75.113 0.750 re f 580.125 280.521 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.262 283.652 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 147.160 268.616 83.753 12.280 re f 0.267 0.267 0.267 rg 147.160 280.521 83.753 0.750 re f 0.271 0.267 0.267 rg BT 151.660 271.372 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 230.913 268.616 71.149 12.280 re f 0.267 0.267 0.267 rg 230.913 280.521 71.149 0.750 re f 0.271 0.267 0.267 rg BT 235.413 271.372 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 302.062 268.616 69.816 12.280 re f 0.267 0.267 0.267 rg 302.062 280.521 69.816 0.750 re f 0.271 0.267 0.267 rg BT 306.562 271.372 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 371.878 268.616 71.149 12.280 re f 0.267 0.267 0.267 rg 371.878 280.521 71.149 0.750 re f 0.965 0.965 0.965 rg 443.026 268.616 62.736 12.280 re f 0.267 0.267 0.267 rg 443.026 280.521 62.736 0.750 re f 0.965 0.965 0.965 rg 505.762 268.616 74.738 12.280 re f 0.267 0.267 0.267 rg 505.762 280.521 75.113 0.750 re f 580.125 268.241 0.750 13.030 re f 0.965 0.965 0.965 rg 147.160 256.336 83.753 12.280 re f 0.267 0.267 0.267 rg 147.160 268.241 83.753 0.750 re f 0.271 0.267 0.267 rg BT 151.660 259.092 Td /F4 9.8 Tf [(F\(1,57\))] TJ ET 0.965 0.965 0.965 rg 230.913 256.336 71.149 12.280 re f 0.267 0.267 0.267 rg 230.913 268.241 71.149 0.750 re f 0.271 0.267 0.267 rg BT 235.413 259.092 Td /F4 9.8 Tf [(F\(3,160\))] TJ ET 0.965 0.965 0.965 rg 302.062 256.336 69.816 12.280 re f 0.267 0.267 0.267 rg 302.062 268.241 69.816 0.750 re f 0.271 0.267 0.267 rg BT 306.562 259.092 Td /F4 9.8 Tf [(F\(1,57\))] TJ ET 0.965 0.965 0.965 rg 371.878 256.336 71.149 12.280 re f 0.267 0.267 0.267 rg 371.878 268.241 71.149 0.750 re f 0.271 0.267 0.267 rg BT 376.378 259.092 Td /F4 9.8 Tf [(F\(3,160\))] TJ ET 0.965 0.965 0.965 rg 443.026 256.336 62.736 12.280 re f 0.267 0.267 0.267 rg 443.026 268.241 62.736 0.750 re f 0.271 0.267 0.267 rg BT 447.526 259.092 Td /F4 9.8 Tf [(F\(1,57\))] TJ ET 0.965 0.965 0.965 rg 505.762 256.336 74.738 12.280 re f 0.267 0.267 0.267 rg 505.762 268.241 75.113 0.750 re f 580.125 255.961 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.262 259.092 Td /F4 9.8 Tf [(F\(6,160\))] TJ ET 0.267 0.267 0.267 rg 26.625 255.961 120.910 0.750 re f 26.625 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 246.106 Td /F4 9.8 Tf [(Corner Entries)] TJ ET 0.267 0.267 0.267 rg 146.785 255.961 84.503 0.750 re f 146.785 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 246.106 Td /F1 9.8 Tf [(42.55#)] TJ ET 0.267 0.267 0.267 rg 230.538 255.961 71.899 0.750 re f 230.538 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 246.106 Td /F1 9.8 Tf [(47.93#)] TJ ET 0.267 0.267 0.267 rg 301.687 255.961 70.566 0.750 re f 301.687 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 246.106 Td /F1 9.8 Tf [(480.78#)] TJ ET 0.267 0.267 0.267 rg 371.503 255.961 71.899 0.750 re f 371.503 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 246.106 Td /F1 9.8 Tf [(7.05#)] TJ ET 0.267 0.267 0.267 rg 442.651 255.961 63.486 0.750 re f 442.651 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 246.106 Td /F1 9.8 Tf [(6.13*)] TJ ET 0.267 0.267 0.267 rg 505.387 255.961 75.488 0.750 re f 505.387 240.080 0.750 16.631 re f 580.125 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 246.106 Td /F1 9.8 Tf [(12.19#)] TJ ET 0.267 0.267 0.267 rg 26.625 240.080 120.910 0.750 re f 26.625 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 230.225 Td /F4 9.8 Tf [(Licking)] TJ ET 0.267 0.267 0.267 rg 146.785 240.080 84.503 0.750 re f 146.785 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 230.225 Td /F1 9.8 Tf [(17.24#)] TJ ET 0.267 0.267 0.267 rg 230.538 240.080 71.899 0.750 re f 230.538 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 230.225 Td /F1 9.8 Tf [(51.31#)] TJ ET 0.267 0.267 0.267 rg 301.687 240.080 70.566 0.750 re f 301.687 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 230.225 Td /F1 9.8 Tf [(168.08#)] TJ ET 0.267 0.267 0.267 rg 371.503 240.080 71.899 0.750 re f 371.503 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 230.225 Td /F1 9.8 Tf [(3.50*)] TJ ET 0.267 0.267 0.267 rg 442.651 240.080 63.486 0.750 re f 442.651 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 230.225 Td /F1 9.8 Tf [(12.62#)] TJ ET 0.267 0.267 0.267 rg 505.387 240.080 75.488 0.750 re f 505.387 224.199 0.750 16.631 re f 580.125 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 230.225 Td /F1 9.8 Tf [(3.39**)] TJ ET 0.267 0.267 0.267 rg 26.625 224.199 120.910 0.750 re f 26.625 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 214.344 Td /F4 9.8 Tf [(Repeats)] TJ ET 0.267 0.267 0.267 rg 146.785 224.199 84.503 0.750 re f 146.785 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 214.344 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 230.538 224.199 71.899 0.750 re f 230.538 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 214.344 Td /F1 9.8 Tf [(52.81#)] TJ ET 0.267 0.267 0.267 rg 301.687 224.199 70.566 0.750 re f 301.687 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 214.344 Td /F1 9.8 Tf [(158.69#)] TJ ET 0.267 0.267 0.267 rg 371.503 224.199 71.899 0.750 re f 371.503 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 214.344 Td /F1 9.8 Tf [(12.04#)] TJ ET 0.267 0.267 0.267 rg 442.651 224.199 63.486 0.750 re f 442.651 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 214.344 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.387 224.199 75.488 0.750 re f 505.387 208.318 0.750 16.631 re f 580.125 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 214.344 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 208.318 120.910 0.750 re f 26.625 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 198.463 Td /F4 9.8 Tf [(Alternations)] TJ ET 0.267 0.267 0.267 rg 146.785 208.318 84.503 0.750 re f 146.785 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 198.463 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 230.538 208.318 71.899 0.750 re f 230.538 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 198.463 Td /F1 9.8 Tf [(6.70#)] TJ ET 0.267 0.267 0.267 rg 301.687 208.318 70.566 0.750 re f 301.687 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 198.463 Td /F1 9.8 Tf [(80.24#)] TJ ET 0.267 0.267 0.267 rg 371.503 208.318 71.899 0.750 re f 371.503 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 198.463 Td /F1 9.8 Tf [(5.07**)] TJ ET 0.267 0.267 0.267 rg 442.651 208.318 63.486 0.750 re f 442.651 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 198.463 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.387 208.318 75.488 0.750 re f 505.387 192.436 0.750 16.631 re f 580.125 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 198.463 Td /F1 9.8 Tf [(9.01#)] TJ ET 0.267 0.267 0.267 rg 26.625 192.436 120.910 0.750 re f 26.625 176.555 120.910 0.750 re f 26.625 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 182.581 Td /F4 9.8 Tf [(Collected Rewards)] TJ ET 0.267 0.267 0.267 rg 146.785 192.436 84.503 0.750 re f 146.785 176.555 84.503 0.750 re f 146.785 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 182.581 Td /F1 9.8 Tf [(43.59#)] TJ ET 0.267 0.267 0.267 rg 230.538 192.436 71.899 0.750 re f 230.538 176.555 71.899 0.750 re f 230.538 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 182.581 Td /F1 9.8 Tf [(32.99#)] TJ ET 0.267 0.267 0.267 rg 301.687 192.436 70.566 0.750 re f 301.687 176.555 70.566 0.750 re f 301.687 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 182.581 Td /F1 9.8 Tf [(31.71#)] TJ ET 0.267 0.267 0.267 rg 371.503 192.436 71.899 0.750 re f 371.503 176.555 71.899 0.750 re f 371.503 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 182.581 Td /F1 9.8 Tf [(17.75#)] TJ ET 0.267 0.267 0.267 rg 442.651 192.436 63.486 0.750 re f 442.651 176.555 63.486 0.750 re f 442.651 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 182.581 Td /F1 9.8 Tf [(4.57*)] TJ ET 0.267 0.267 0.267 rg 505.387 192.436 75.488 0.750 re f 505.387 176.555 75.488 0.750 re f 505.387 176.555 0.750 16.631 re f 580.125 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 182.581 Td /F1 9.8 Tf [(NS)] TJ ET 0.965 0.965 0.965 rg 26.250 -334.570 555.000 465.750 re f 0.267 0.267 0.267 rg 26.250 131.180 m 581.250 131.180 l 581.250 130.430 l 26.250 130.430 l f q 278.250 0 0 450.000 35.250 -328.570 cm /I9 Do Q q 35.250 -334.570 537.000 0.000 re W n Q Q q 15.000 -334.570 577.500 1111.570 re W n 0.965 0.965 0.965 rg 26.250 700.968 555.000 76.032 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 700.968 m 581.250 700.968 l 581.250 701.718 l 26.250 701.718 l f q 35.250 712.218 537.000 64.782 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 6: Behavioral measures derived from the computer vision of Experiment 1 \(R6/2\) as a function of genotype, )] TJ ET BT 35.250 755.571 Td /F4 9.8 Tf [(age, and light cycle.)] TJ ET BT 35.250 736.201 Td /F1 9.8 Tf [(***: significant genotype effect, independent of cycle or age; ##: significant differences due to light cycle for each age )] TJ ET BT 35.250 722.465 Td /F1 9.8 Tf [(independently of genotype.)] TJ ET Q BT 26.250 683.944 Td /F4 9.8 Tf [(Experiment 2. Analysis of BACHD mice:)] TJ ET BT 26.250 664.539 Td /F5 9.8 Tf [(Corner Entries:)] TJ ET BT 91.809 664.539 Td /F1 9.8 Tf [( BACHD mice exhibited a different phenotype than R6/2 mice in terms of the degree of corner visits: R6/2 mice )] TJ ET BT 26.250 652.635 Td /F1 9.8 Tf [(did not differ from their WT controls in terms of the frequency of corner visits however, BACHD mice entered the corners )] TJ ET BT 26.250 640.730 Td /F1 9.8 Tf [(significantly less than WT mice at all ages and cycle phases with the exception of the dark phase at 16 weeks of age \(Figure )] TJ ET BT 26.250 628.825 Td /F1 9.8 Tf [(7A; Table 3; Genotype x Age x Cycle interaction: )] TJ ET BT 239.229 628.825 Td /F5 9.8 Tf [(p)] TJ ET BT 244.650 628.825 Td /F1 9.8 Tf [(< 001; simple main effects: )] TJ ET BT 363.600 628.825 Td /F5 9.8 Tf [(p)] TJ ET BT 369.021 628.825 Td /F1 9.8 Tf [(s < 0.05\). Both groups were more active during )] TJ ET BT 26.250 616.920 Td /F1 9.8 Tf [(the dark period \(Cycle main effect: )] TJ ET BT 177.434 616.920 Td /F5 9.8 Tf [(p)] TJ ET BT 182.855 616.920 Td /F1 9.8 Tf [(< 0.001\).)] TJ ET BT 26.250 597.516 Td /F5 9.8 Tf [(Licking:)] TJ ET BT 59.303 597.516 Td /F1 9.8 Tf [( Similar to the phenotype of R6/2 mice, BACHD mice exhibited increased thirst, which was not detected at the older )] TJ ET BT 26.250 585.611 Td /F1 9.8 Tf [(ages \(i.e., 10 weeks of age for R6/2 mice and 52 weeks of age for BACHD mice\). At earlier ages, the BACHD mice licked more )] TJ ET BT 26.250 573.706 Td /F1 9.8 Tf [(than did WT mice irrespective of cycle phase, but this difference waned as testing progressed \(Figure 7B; Table 3; Genotype x )] TJ ET BT 26.250 561.801 Td /F1 9.8 Tf [(Age x Cycle interaction: )] TJ ET BT 131.375 561.801 Td /F5 9.8 Tf [(p)] TJ ET BT 136.796 561.801 Td /F1 9.8 Tf [( < 0.01; simple main effects: )] TJ ET BT 261.166 561.801 Td /F5 9.8 Tf [(p)] TJ ET BT 266.587 561.801 Td /F1 9.8 Tf [(s < 0.05\). As expected, both groups licked more during the night period )] TJ ET BT 26.250 549.897 Td /F1 9.8 Tf [(\(with the exemption of WT mice at 36 weeks of age\) and, unexpectedly, licked less as testing progressed \(Age main effect: )] TJ ET BT 557.859 549.897 Td /F5 9.8 Tf [(p)] TJ ET BT 563.280 549.897 Td /F1 9.8 Tf [( < )] TJ ET BT 26.250 537.992 Td /F1 9.8 Tf [(0.001\).)] TJ ET BT 26.250 518.587 Td /F5 9.8 Tf [(Repeat visits:)] TJ ET BT 84.769 518.587 Td /F1 9.8 Tf [( Perseverative responding was apparent only during early test ages for BACHD mice. BACHD mice re-entered )] TJ ET BT 26.250 506.682 Td /F1 9.8 Tf [(previously visited corners more frequently than did WT controls at 16 and 24 weeks of age but less at 52 weeks of age \(Figure )] TJ ET BT 26.250 494.778 Td /F1 9.8 Tf [(7C; Table 3; Genotype x Age interaction: )] TJ ET BT 205.094 494.778 Td /F5 9.8 Tf [(p)] TJ ET BT 210.515 494.778 Td /F1 9.8 Tf [( < 0.001; simple main effects: )] TJ ET BT 340.307 494.778 Td /F5 9.8 Tf [(p)] TJ ET BT 345.728 494.778 Td /F1 9.8 Tf [(s < 0.05\). Both groups made more repeats when they )] TJ ET BT 26.250 482.873 Td /F1 9.8 Tf [(were active during the dark phase and made fewer repeats as testing continued \(Age and Cycle main effect: )] TJ ET BT 495.020 482.873 Td /F5 9.8 Tf [(p)] TJ ET BT 500.441 482.873 Td /F1 9.8 Tf [(s < 0.001\).)] TJ ET BT 26.250 463.468 Td /F5 9.8 Tf [(Alternation. )] TJ ET BT 78.276 463.468 Td /F1 9.8 Tf [(Similar age-dependent patterns of genotypic differences were also observed with spatial alternation performance. )] TJ ET BT 26.250 451.563 Td /F1 9.8 Tf [(BACHD mice alternated less than WT during both cycle phases at 16 weeks of age but the effect was not apparent at later )] TJ ET BT 26.250 439.659 Td /F1 9.8 Tf [(ages. BACHD mice alternated more during the light phase at all ages, while this difference was only observed in WT controls at )] TJ ET BT 26.250 427.754 Td /F1 9.8 Tf [(16 weeks of age. \(Figure 7D; Table 3; Genotype x Age x Cycle interaction: )] TJ ET BT 349.774 427.754 Td /F5 9.8 Tf [(p)] TJ ET BT 355.195 427.754 Td /F1 9.8 Tf [( < 0.001\).)] TJ ET BT 26.250 408.349 Td /F5 9.8 Tf [(Rewards collected: )] TJ ET BT 111.319 408.349 Td /F1 9.8 Tf [(Interestingly, BACHD mice exhibited higher efficiency of reward collection than WT animals, a characteristic )] TJ ET BT 26.250 396.444 Td /F1 9.8 Tf [(that became more pronounced with age. For instance, starting at 24 weeks of age BACHD mice were more likely than WT mice )] TJ ET BT 26.250 384.540 Td /F1 9.8 Tf [(to nosepoke in active corners. Both groups, surprisingly, were slightly but consistently less likely to collect rewards during the )] TJ ET BT 26.250 372.635 Td /F1 9.8 Tf [(dark phase, particularly the BACHD mice \(Figure 7E; Table 3; Genotype x Age and Genotype x Cycle interactions: )] TJ ET BT 521.540 372.635 Td /F5 9.8 Tf [(p)] TJ ET BT 526.961 372.635 Td /F1 9.8 Tf [(s < 0.05; )] TJ ET BT 26.250 360.730 Td /F1 9.8 Tf [(simple main effect: )] TJ ET BT 110.237 360.730 Td /F5 9.8 Tf [(p)] TJ ET BT 115.658 360.730 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET q 26.250 337.113 555.000 13.736 re W n 0.271 0.267 0.267 rg BT 26.250 339.860 Td /F1 9.8 Tf [(Table 3. Summary of the statistical analyses for individual behavioral measures for Experiment 2 \(BACHD\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 306.205 555.000 23.408 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 329.613 m 581.250 329.613 l 580.500 328.863 l 27.000 328.863 l f 581.250 329.613 m 581.250 306.205 l 580.500 306.205 l 580.500 328.863 l f 26.250 329.613 m 26.250 306.205 l 27.000 306.205 l 27.000 328.863 l f 0.271 0.267 0.267 rg BT 33.000 315.537 Td /F1 9.0 Tf [(Notes: *: p< 0.05, **: p< 0.01, #: p< 0.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 176.180 555.000 130.025 re f 0.965 0.965 0.965 rg 27.000 256.336 120.160 49.119 re f 0.267 0.267 0.267 rg 26.625 305.080 120.535 0.750 re f 26.625 293.176 0.750 12.655 re f 26.625 280.896 0.750 12.280 re f 26.625 268.616 0.750 12.280 re f 26.625 255.961 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 295.932 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 147.160 293.176 433.340 12.280 re f 0.267 0.267 0.267 rg 147.160 305.080 83.753 0.750 re f 230.913 305.080 71.149 0.750 re f 302.062 305.080 69.816 0.750 re f 371.878 305.080 71.149 0.750 re f 443.026 305.080 62.736 0.750 re f 505.762 305.080 75.113 0.750 re f 580.125 292.801 0.750 13.030 re f 0.271 0.267 0.267 rg BT 151.660 295.932 Td /F4 9.8 Tf [(BACHD)] TJ ET 0.965 0.965 0.965 rg 147.160 280.896 83.753 12.280 re f 0.267 0.267 0.267 rg 147.160 292.801 83.753 0.750 re f 0.271 0.267 0.267 rg BT 151.660 283.652 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 230.913 280.896 71.149 12.280 re f 0.267 0.267 0.267 rg 230.913 292.801 71.149 0.750 re f 0.271 0.267 0.267 rg BT 235.413 283.652 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 302.062 280.896 69.816 12.280 re f 0.267 0.267 0.267 rg 302.062 292.801 69.816 0.750 re f 0.271 0.267 0.267 rg BT 306.562 283.652 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 371.878 280.896 71.149 12.280 re f 0.267 0.267 0.267 rg 371.878 292.801 71.149 0.750 re f 0.271 0.267 0.267 rg BT 376.378 283.652 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 443.026 280.896 62.736 12.280 re f 0.267 0.267 0.267 rg 443.026 292.801 62.736 0.750 re f 0.271 0.267 0.267 rg BT 447.526 283.652 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 505.762 280.896 74.738 12.280 re f 0.267 0.267 0.267 rg 505.762 292.801 75.113 0.750 re f 580.125 280.521 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.262 283.652 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 147.160 268.616 83.753 12.280 re f 0.267 0.267 0.267 rg 147.160 280.521 83.753 0.750 re f 0.271 0.267 0.267 rg BT 151.660 271.372 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 230.913 268.616 71.149 12.280 re f 0.267 0.267 0.267 rg 230.913 280.521 71.149 0.750 re f 0.271 0.267 0.267 rg BT 235.413 271.372 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 302.062 268.616 69.816 12.280 re f 0.267 0.267 0.267 rg 302.062 280.521 69.816 0.750 re f 0.271 0.267 0.267 rg BT 306.562 271.372 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 371.878 268.616 71.149 12.280 re f 0.267 0.267 0.267 rg 371.878 280.521 71.149 0.750 re f 0.965 0.965 0.965 rg 443.026 268.616 62.736 12.280 re f 0.267 0.267 0.267 rg 443.026 280.521 62.736 0.750 re f 0.965 0.965 0.965 rg 505.762 268.616 74.738 12.280 re f 0.267 0.267 0.267 rg 505.762 280.521 75.113 0.750 re f 580.125 268.241 0.750 13.030 re f 0.965 0.965 0.965 rg 147.160 256.336 83.753 12.280 re f 0.267 0.267 0.267 rg 147.160 268.241 83.753 0.750 re f 0.271 0.267 0.267 rg BT 151.660 259.092 Td /F4 9.8 Tf [(F\(1,57\))] TJ ET 0.965 0.965 0.965 rg 230.913 256.336 71.149 12.280 re f 0.267 0.267 0.267 rg 230.913 268.241 71.149 0.750 re f 0.271 0.267 0.267 rg BT 235.413 259.092 Td /F4 9.8 Tf [(F\(3,160\))] TJ ET 0.965 0.965 0.965 rg 302.062 256.336 69.816 12.280 re f 0.267 0.267 0.267 rg 302.062 268.241 69.816 0.750 re f 0.271 0.267 0.267 rg BT 306.562 259.092 Td /F4 9.8 Tf [(F\(1,57\))] TJ ET 0.965 0.965 0.965 rg 371.878 256.336 71.149 12.280 re f 0.267 0.267 0.267 rg 371.878 268.241 71.149 0.750 re f 0.271 0.267 0.267 rg BT 376.378 259.092 Td /F4 9.8 Tf [(F\(3,160\))] TJ ET 0.965 0.965 0.965 rg 443.026 256.336 62.736 12.280 re f 0.267 0.267 0.267 rg 443.026 268.241 62.736 0.750 re f 0.271 0.267 0.267 rg BT 447.526 259.092 Td /F4 9.8 Tf [(F\(1,57\))] TJ ET 0.965 0.965 0.965 rg 505.762 256.336 74.738 12.280 re f 0.267 0.267 0.267 rg 505.762 268.241 75.113 0.750 re f 580.125 255.961 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.262 259.092 Td /F4 9.8 Tf [(F\(6,160\))] TJ ET 0.267 0.267 0.267 rg 26.625 255.961 120.910 0.750 re f 26.625 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 246.106 Td /F4 9.8 Tf [(Corner Entries)] TJ ET 0.267 0.267 0.267 rg 146.785 255.961 84.503 0.750 re f 146.785 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 246.106 Td /F1 9.8 Tf [(42.55#)] TJ ET 0.267 0.267 0.267 rg 230.538 255.961 71.899 0.750 re f 230.538 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 246.106 Td /F1 9.8 Tf [(47.93#)] TJ ET 0.267 0.267 0.267 rg 301.687 255.961 70.566 0.750 re f 301.687 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 246.106 Td /F1 9.8 Tf [(480.78#)] TJ ET 0.267 0.267 0.267 rg 371.503 255.961 71.899 0.750 re f 371.503 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 246.106 Td /F1 9.8 Tf [(7.05#)] TJ ET 0.267 0.267 0.267 rg 442.651 255.961 63.486 0.750 re f 442.651 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 246.106 Td /F1 9.8 Tf [(6.13*)] TJ ET 0.267 0.267 0.267 rg 505.387 255.961 75.488 0.750 re f 505.387 240.080 0.750 16.631 re f 580.125 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 246.106 Td /F1 9.8 Tf [(12.19#)] TJ ET 0.267 0.267 0.267 rg 26.625 240.080 120.910 0.750 re f 26.625 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 230.225 Td /F4 9.8 Tf [(Licking)] TJ ET 0.267 0.267 0.267 rg 146.785 240.080 84.503 0.750 re f 146.785 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 230.225 Td /F1 9.8 Tf [(17.24#)] TJ ET 0.267 0.267 0.267 rg 230.538 240.080 71.899 0.750 re f 230.538 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 230.225 Td /F1 9.8 Tf [(51.31#)] TJ ET 0.267 0.267 0.267 rg 301.687 240.080 70.566 0.750 re f 301.687 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 230.225 Td /F1 9.8 Tf [(168.08#)] TJ ET 0.267 0.267 0.267 rg 371.503 240.080 71.899 0.750 re f 371.503 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 230.225 Td /F1 9.8 Tf [(3.50*)] TJ ET 0.267 0.267 0.267 rg 442.651 240.080 63.486 0.750 re f 442.651 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 230.225 Td /F1 9.8 Tf [(12.62#)] TJ ET 0.267 0.267 0.267 rg 505.387 240.080 75.488 0.750 re f 505.387 224.199 0.750 16.631 re f 580.125 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 230.225 Td /F1 9.8 Tf [(3.39**)] TJ ET 0.267 0.267 0.267 rg 26.625 224.199 120.910 0.750 re f 26.625 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 214.344 Td /F4 9.8 Tf [(Repeats)] TJ ET 0.267 0.267 0.267 rg 146.785 224.199 84.503 0.750 re f 146.785 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 214.344 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 230.538 224.199 71.899 0.750 re f 230.538 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 214.344 Td /F1 9.8 Tf [(52.81#)] TJ ET 0.267 0.267 0.267 rg 301.687 224.199 70.566 0.750 re f 301.687 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 214.344 Td /F1 9.8 Tf [(158.69#)] TJ ET 0.267 0.267 0.267 rg 371.503 224.199 71.899 0.750 re f 371.503 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 214.344 Td /F1 9.8 Tf [(12.04#)] TJ ET 0.267 0.267 0.267 rg 442.651 224.199 63.486 0.750 re f 442.651 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 214.344 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.387 224.199 75.488 0.750 re f 505.387 208.318 0.750 16.631 re f 580.125 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 214.344 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 208.318 120.910 0.750 re f 26.625 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 198.463 Td /F4 9.8 Tf [(Alternations)] TJ ET 0.267 0.267 0.267 rg 146.785 208.318 84.503 0.750 re f 146.785 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 198.463 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 230.538 208.318 71.899 0.750 re f 230.538 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 198.463 Td /F1 9.8 Tf [(6.70#)] TJ ET 0.267 0.267 0.267 rg 301.687 208.318 70.566 0.750 re f 301.687 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 198.463 Td /F1 9.8 Tf [(80.24#)] TJ ET 0.267 0.267 0.267 rg 371.503 208.318 71.899 0.750 re f 371.503 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 198.463 Td /F1 9.8 Tf [(5.07**)] TJ ET 0.267 0.267 0.267 rg 442.651 208.318 63.486 0.750 re f 442.651 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 198.463 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.387 208.318 75.488 0.750 re f 505.387 192.436 0.750 16.631 re f 580.125 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 198.463 Td /F1 9.8 Tf [(9.01#)] TJ ET 0.267 0.267 0.267 rg 26.625 192.436 120.910 0.750 re f 26.625 176.555 120.910 0.750 re f 26.625 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 182.581 Td /F4 9.8 Tf [(Collected Rewards)] TJ ET 0.267 0.267 0.267 rg 146.785 192.436 84.503 0.750 re f 146.785 176.555 84.503 0.750 re f 146.785 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 182.581 Td /F1 9.8 Tf [(43.59#)] TJ ET 0.267 0.267 0.267 rg 230.538 192.436 71.899 0.750 re f 230.538 176.555 71.899 0.750 re f 230.538 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 182.581 Td /F1 9.8 Tf [(32.99#)] TJ ET 0.267 0.267 0.267 rg 301.687 192.436 70.566 0.750 re f 301.687 176.555 70.566 0.750 re f 301.687 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 182.581 Td /F1 9.8 Tf [(31.71#)] TJ ET 0.267 0.267 0.267 rg 371.503 192.436 71.899 0.750 re f 371.503 176.555 71.899 0.750 re f 371.503 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 182.581 Td /F1 9.8 Tf [(17.75#)] TJ ET 0.267 0.267 0.267 rg 442.651 192.436 63.486 0.750 re f 442.651 176.555 63.486 0.750 re f 442.651 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 182.581 Td /F1 9.8 Tf [(4.57*)] TJ ET 0.267 0.267 0.267 rg 505.387 192.436 75.488 0.750 re f 505.387 176.555 75.488 0.750 re f 505.387 176.555 0.750 16.631 re f 580.125 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 182.581 Td /F1 9.8 Tf [(NS)] TJ ET 0.965 0.965 0.965 rg 26.250 -334.570 555.000 465.750 re f 0.267 0.267 0.267 rg 26.250 131.180 m 581.250 131.180 l 581.250 130.430 l 26.250 130.430 l f q 278.250 0 0 450.000 35.250 -328.570 cm /I9 Do Q q 35.250 -334.570 537.000 0.000 re W n Q Q q 15.000 -334.570 577.500 1111.570 re W n 0.965 0.965 0.965 rg 26.250 700.968 555.000 76.032 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 700.968 m 581.250 700.968 l 581.250 701.718 l 26.250 701.718 l f q 35.250 712.218 537.000 64.782 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 6: Behavioral measures derived from the computer vision of Experiment 1 \(R6/2\) as a function of genotype, )] TJ ET BT 35.250 755.571 Td /F4 9.8 Tf [(age, and light cycle.)] TJ ET BT 35.250 736.201 Td /F1 9.8 Tf [(***: significant genotype effect, independent of cycle or age; ##: significant differences due to light cycle for each age )] TJ ET BT 35.250 722.465 Td /F1 9.8 Tf [(independently of genotype.)] TJ ET Q BT 26.250 683.944 Td /F4 9.8 Tf [(Experiment 2. Analysis of BACHD mice:)] TJ ET BT 26.250 664.539 Td /F5 9.8 Tf [(Corner Entries:)] TJ ET BT 91.809 664.539 Td /F1 9.8 Tf [( BACHD mice exhibited a different phenotype than R6/2 mice in terms of the degree of corner visits: R6/2 mice )] TJ ET BT 26.250 652.635 Td /F1 9.8 Tf [(did not differ from their WT controls in terms of the frequency of corner visits however, BACHD mice entered the corners )] TJ ET BT 26.250 640.730 Td /F1 9.8 Tf [(significantly less than WT mice at all ages and cycle phases with the exception of the dark phase at 16 weeks of age \(Figure )] TJ ET BT 26.250 628.825 Td /F1 9.8 Tf [(7A; Table 3; Genotype x Age x Cycle interaction: )] TJ ET BT 239.229 628.825 Td /F5 9.8 Tf [(p)] TJ ET BT 244.650 628.825 Td /F1 9.8 Tf [(< 001; simple main effects: )] TJ ET BT 363.600 628.825 Td /F5 9.8 Tf [(p)] TJ ET BT 369.021 628.825 Td /F1 9.8 Tf [(s < 0.05\). Both groups were more active during )] TJ ET BT 26.250 616.920 Td /F1 9.8 Tf [(the dark period \(Cycle main effect: )] TJ ET BT 177.434 616.920 Td /F5 9.8 Tf [(p)] TJ ET BT 182.855 616.920 Td /F1 9.8 Tf [(< 0.001\).)] TJ ET BT 26.250 597.516 Td /F5 9.8 Tf [(Licking:)] TJ ET BT 59.303 597.516 Td /F1 9.8 Tf [( Similar to the phenotype of R6/2 mice, BACHD mice exhibited increased thirst, which was not detected at the older )] TJ ET BT 26.250 585.611 Td /F1 9.8 Tf [(ages \(i.e., 10 weeks of age for R6/2 mice and 52 weeks of age for BACHD mice\). At earlier ages, the BACHD mice licked more )] TJ ET BT 26.250 573.706 Td /F1 9.8 Tf [(than did WT mice irrespective of cycle phase, but this difference waned as testing progressed \(Figure 7B; Table 3; Genotype x )] TJ ET BT 26.250 561.801 Td /F1 9.8 Tf [(Age x Cycle interaction: )] TJ ET BT 131.375 561.801 Td /F5 9.8 Tf [(p)] TJ ET BT 136.796 561.801 Td /F1 9.8 Tf [( < 0.01; simple main effects: )] TJ ET BT 261.166 561.801 Td /F5 9.8 Tf [(p)] TJ ET BT 266.587 561.801 Td /F1 9.8 Tf [(s < 0.05\). As expected, both groups licked more during the night period )] TJ ET BT 26.250 549.897 Td /F1 9.8 Tf [(\(with the exemption of WT mice at 36 weeks of age\) and, unexpectedly, licked less as testing progressed \(Age main effect: )] TJ ET BT 557.859 549.897 Td /F5 9.8 Tf [(p)] TJ ET BT 563.280 549.897 Td /F1 9.8 Tf [( < )] TJ ET BT 26.250 537.992 Td /F1 9.8 Tf [(0.001\).)] TJ ET BT 26.250 518.587 Td /F5 9.8 Tf [(Repeat visits:)] TJ ET BT 84.769 518.587 Td /F1 9.8 Tf [( Perseverative responding was apparent only during early test ages for BACHD mice. BACHD mice re-entered )] TJ ET BT 26.250 506.682 Td /F1 9.8 Tf [(previously visited corners more frequently than did WT controls at 16 and 24 weeks of age but less at 52 weeks of age \(Figure )] TJ ET BT 26.250 494.778 Td /F1 9.8 Tf [(7C; Table 3; Genotype x Age interaction: )] TJ ET BT 205.094 494.778 Td /F5 9.8 Tf [(p)] TJ ET BT 210.515 494.778 Td /F1 9.8 Tf [( < 0.001; simple main effects: )] TJ ET BT 340.307 494.778 Td /F5 9.8 Tf [(p)] TJ ET BT 345.728 494.778 Td /F1 9.8 Tf [(s < 0.05\). Both groups made more repeats when they )] TJ ET BT 26.250 482.873 Td /F1 9.8 Tf [(were active during the dark phase and made fewer repeats as testing continued \(Age and Cycle main effect: )] TJ ET BT 495.020 482.873 Td /F5 9.8 Tf [(p)] TJ ET BT 500.441 482.873 Td /F1 9.8 Tf [(s < 0.001\).)] TJ ET BT 26.250 463.468 Td /F5 9.8 Tf [(Alternation. )] TJ ET BT 78.276 463.468 Td /F1 9.8 Tf [(Similar age-dependent patterns of genotypic differences were also observed with spatial alternation performance. )] TJ ET BT 26.250 451.563 Td /F1 9.8 Tf [(BACHD mice alternated less than WT during both cycle phases at 16 weeks of age but the effect was not apparent at later )] TJ ET BT 26.250 439.659 Td /F1 9.8 Tf [(ages. BACHD mice alternated more during the light phase at all ages, while this difference was only observed in WT controls at )] TJ ET BT 26.250 427.754 Td /F1 9.8 Tf [(16 weeks of age. \(Figure 7D; Table 3; Genotype x Age x Cycle interaction: )] TJ ET BT 349.774 427.754 Td /F5 9.8 Tf [(p)] TJ ET BT 355.195 427.754 Td /F1 9.8 Tf [( < 0.001\).)] TJ ET BT 26.250 408.349 Td /F5 9.8 Tf [(Rewards collected: )] TJ ET BT 111.319 408.349 Td /F1 9.8 Tf [(Interestingly, BACHD mice exhibited higher efficiency of reward collection than WT animals, a characteristic )] TJ ET BT 26.250 396.444 Td /F1 9.8 Tf [(that became more pronounced with age. For instance, starting at 24 weeks of age BACHD mice were more likely than WT mice )] TJ ET BT 26.250 384.540 Td /F1 9.8 Tf [(to nosepoke in active corners. Both groups, surprisingly, were slightly but consistently less likely to collect rewards during the )] TJ ET BT 26.250 372.635 Td /F1 9.8 Tf [(dark phase, particularly the BACHD mice \(Figure 7E; Table 3; Genotype x Age and Genotype x Cycle interactions: )] TJ ET BT 521.540 372.635 Td /F5 9.8 Tf [(p)] TJ ET BT 526.961 372.635 Td /F1 9.8 Tf [(s < 0.05; )] TJ ET BT 26.250 360.730 Td /F1 9.8 Tf [(simple main effect: )] TJ ET BT 110.237 360.730 Td /F5 9.8 Tf [(p)] TJ ET BT 115.658 360.730 Td /F1 9.8 Tf [(s < 0.05\).)] TJ ET q 26.250 337.113 555.000 13.736 re W n 0.271 0.267 0.267 rg BT 26.250 339.860 Td /F1 9.8 Tf [(Table 3. Summary of the statistical analyses for individual behavioral measures for Experiment 2 \(BACHD\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 306.205 555.000 23.408 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 329.613 m 581.250 329.613 l 580.500 328.863 l 27.000 328.863 l f 581.250 329.613 m 581.250 306.205 l 580.500 306.205 l 580.500 328.863 l f 26.250 329.613 m 26.250 306.205 l 27.000 306.205 l 27.000 328.863 l f 0.271 0.267 0.267 rg BT 33.000 315.537 Td /F1 9.0 Tf [(Notes: *: p< 0.05, **: p< 0.01, #: p< 0.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 176.180 555.000 130.025 re f 0.965 0.965 0.965 rg 27.000 256.336 120.160 49.119 re f 0.267 0.267 0.267 rg 26.625 305.080 120.535 0.750 re f 26.625 293.176 0.750 12.655 re f 26.625 280.896 0.750 12.280 re f 26.625 268.616 0.750 12.280 re f 26.625 255.961 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 295.932 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 147.160 293.176 433.340 12.280 re f 0.267 0.267 0.267 rg 147.160 305.080 83.753 0.750 re f 230.913 305.080 71.149 0.750 re f 302.062 305.080 69.816 0.750 re f 371.878 305.080 71.149 0.750 re f 443.026 305.080 62.736 0.750 re f 505.762 305.080 75.113 0.750 re f 580.125 292.801 0.750 13.030 re f 0.271 0.267 0.267 rg BT 151.660 295.932 Td /F4 9.8 Tf [(BACHD)] TJ ET 0.965 0.965 0.965 rg 147.160 280.896 83.753 12.280 re f 0.267 0.267 0.267 rg 147.160 292.801 83.753 0.750 re f 0.271 0.267 0.267 rg BT 151.660 283.652 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 230.913 280.896 71.149 12.280 re f 0.267 0.267 0.267 rg 230.913 292.801 71.149 0.750 re f 0.271 0.267 0.267 rg BT 235.413 283.652 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 302.062 280.896 69.816 12.280 re f 0.267 0.267 0.267 rg 302.062 292.801 69.816 0.750 re f 0.271 0.267 0.267 rg BT 306.562 283.652 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 371.878 280.896 71.149 12.280 re f 0.267 0.267 0.267 rg 371.878 292.801 71.149 0.750 re f 0.271 0.267 0.267 rg BT 376.378 283.652 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 443.026 280.896 62.736 12.280 re f 0.267 0.267 0.267 rg 443.026 292.801 62.736 0.750 re f 0.271 0.267 0.267 rg BT 447.526 283.652 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 505.762 280.896 74.738 12.280 re f 0.267 0.267 0.267 rg 505.762 292.801 75.113 0.750 re f 580.125 280.521 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.262 283.652 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 147.160 268.616 83.753 12.280 re f 0.267 0.267 0.267 rg 147.160 280.521 83.753 0.750 re f 0.271 0.267 0.267 rg BT 151.660 271.372 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 230.913 268.616 71.149 12.280 re f 0.267 0.267 0.267 rg 230.913 280.521 71.149 0.750 re f 0.271 0.267 0.267 rg BT 235.413 271.372 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 302.062 268.616 69.816 12.280 re f 0.267 0.267 0.267 rg 302.062 280.521 69.816 0.750 re f 0.271 0.267 0.267 rg BT 306.562 271.372 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 371.878 268.616 71.149 12.280 re f 0.267 0.267 0.267 rg 371.878 280.521 71.149 0.750 re f 0.965 0.965 0.965 rg 443.026 268.616 62.736 12.280 re f 0.267 0.267 0.267 rg 443.026 280.521 62.736 0.750 re f 0.965 0.965 0.965 rg 505.762 268.616 74.738 12.280 re f 0.267 0.267 0.267 rg 505.762 280.521 75.113 0.750 re f 580.125 268.241 0.750 13.030 re f 0.965 0.965 0.965 rg 147.160 256.336 83.753 12.280 re f 0.267 0.267 0.267 rg 147.160 268.241 83.753 0.750 re f 0.271 0.267 0.267 rg BT 151.660 259.092 Td /F4 9.8 Tf [(F\(1,57\))] TJ ET 0.965 0.965 0.965 rg 230.913 256.336 71.149 12.280 re f 0.267 0.267 0.267 rg 230.913 268.241 71.149 0.750 re f 0.271 0.267 0.267 rg BT 235.413 259.092 Td /F4 9.8 Tf [(F\(3,160\))] TJ ET 0.965 0.965 0.965 rg 302.062 256.336 69.816 12.280 re f 0.267 0.267 0.267 rg 302.062 268.241 69.816 0.750 re f 0.271 0.267 0.267 rg BT 306.562 259.092 Td /F4 9.8 Tf [(F\(1,57\))] TJ ET 0.965 0.965 0.965 rg 371.878 256.336 71.149 12.280 re f 0.267 0.267 0.267 rg 371.878 268.241 71.149 0.750 re f 0.271 0.267 0.267 rg BT 376.378 259.092 Td /F4 9.8 Tf [(F\(3,160\))] TJ ET 0.965 0.965 0.965 rg 443.026 256.336 62.736 12.280 re f 0.267 0.267 0.267 rg 443.026 268.241 62.736 0.750 re f 0.271 0.267 0.267 rg BT 447.526 259.092 Td /F4 9.8 Tf [(F\(1,57\))] TJ ET 0.965 0.965 0.965 rg 505.762 256.336 74.738 12.280 re f 0.267 0.267 0.267 rg 505.762 268.241 75.113 0.750 re f 580.125 255.961 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.262 259.092 Td /F4 9.8 Tf [(F\(6,160\))] TJ ET 0.267 0.267 0.267 rg 26.625 255.961 120.910 0.750 re f 26.625 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 246.106 Td /F4 9.8 Tf [(Corner Entries)] TJ ET 0.267 0.267 0.267 rg 146.785 255.961 84.503 0.750 re f 146.785 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 246.106 Td /F1 9.8 Tf [(42.55#)] TJ ET 0.267 0.267 0.267 rg 230.538 255.961 71.899 0.750 re f 230.538 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 246.106 Td /F1 9.8 Tf [(47.93#)] TJ ET 0.267 0.267 0.267 rg 301.687 255.961 70.566 0.750 re f 301.687 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 246.106 Td /F1 9.8 Tf [(480.78#)] TJ ET 0.267 0.267 0.267 rg 371.503 255.961 71.899 0.750 re f 371.503 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 246.106 Td /F1 9.8 Tf [(7.05#)] TJ ET 0.267 0.267 0.267 rg 442.651 255.961 63.486 0.750 re f 442.651 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 246.106 Td /F1 9.8 Tf [(6.13*)] TJ ET 0.267 0.267 0.267 rg 505.387 255.961 75.488 0.750 re f 505.387 240.080 0.750 16.631 re f 580.125 240.080 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 246.106 Td /F1 9.8 Tf [(12.19#)] TJ ET 0.267 0.267 0.267 rg 26.625 240.080 120.910 0.750 re f 26.625 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 230.225 Td /F4 9.8 Tf [(Licking)] TJ ET 0.267 0.267 0.267 rg 146.785 240.080 84.503 0.750 re f 146.785 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 230.225 Td /F1 9.8 Tf [(17.24#)] TJ ET 0.267 0.267 0.267 rg 230.538 240.080 71.899 0.750 re f 230.538 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 230.225 Td /F1 9.8 Tf [(51.31#)] TJ ET 0.267 0.267 0.267 rg 301.687 240.080 70.566 0.750 re f 301.687 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 230.225 Td /F1 9.8 Tf [(168.08#)] TJ ET 0.267 0.267 0.267 rg 371.503 240.080 71.899 0.750 re f 371.503 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 230.225 Td /F1 9.8 Tf [(3.50*)] TJ ET 0.267 0.267 0.267 rg 442.651 240.080 63.486 0.750 re f 442.651 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 230.225 Td /F1 9.8 Tf [(12.62#)] TJ ET 0.267 0.267 0.267 rg 505.387 240.080 75.488 0.750 re f 505.387 224.199 0.750 16.631 re f 580.125 224.199 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 230.225 Td /F1 9.8 Tf [(3.39**)] TJ ET 0.267 0.267 0.267 rg 26.625 224.199 120.910 0.750 re f 26.625 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 214.344 Td /F4 9.8 Tf [(Repeats)] TJ ET 0.267 0.267 0.267 rg 146.785 224.199 84.503 0.750 re f 146.785 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 214.344 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 230.538 224.199 71.899 0.750 re f 230.538 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 214.344 Td /F1 9.8 Tf [(52.81#)] TJ ET 0.267 0.267 0.267 rg 301.687 224.199 70.566 0.750 re f 301.687 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 214.344 Td /F1 9.8 Tf [(158.69#)] TJ ET 0.267 0.267 0.267 rg 371.503 224.199 71.899 0.750 re f 371.503 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 214.344 Td /F1 9.8 Tf [(12.04#)] TJ ET 0.267 0.267 0.267 rg 442.651 224.199 63.486 0.750 re f 442.651 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 214.344 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.387 224.199 75.488 0.750 re f 505.387 208.318 0.750 16.631 re f 580.125 208.318 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 214.344 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 208.318 120.910 0.750 re f 26.625 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 198.463 Td /F4 9.8 Tf [(Alternations)] TJ ET 0.267 0.267 0.267 rg 146.785 208.318 84.503 0.750 re f 146.785 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 198.463 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 230.538 208.318 71.899 0.750 re f 230.538 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 198.463 Td /F1 9.8 Tf [(6.70#)] TJ ET 0.267 0.267 0.267 rg 301.687 208.318 70.566 0.750 re f 301.687 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 198.463 Td /F1 9.8 Tf [(80.24#)] TJ ET 0.267 0.267 0.267 rg 371.503 208.318 71.899 0.750 re f 371.503 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 198.463 Td /F1 9.8 Tf [(5.07**)] TJ ET 0.267 0.267 0.267 rg 442.651 208.318 63.486 0.750 re f 442.651 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 198.463 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.387 208.318 75.488 0.750 re f 505.387 192.436 0.750 16.631 re f 580.125 192.436 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 198.463 Td /F1 9.8 Tf [(9.01#)] TJ ET 0.267 0.267 0.267 rg 26.625 192.436 120.910 0.750 re f 26.625 176.555 120.910 0.750 re f 26.625 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 182.581 Td /F4 9.8 Tf [(Collected Rewards)] TJ ET 0.267 0.267 0.267 rg 146.785 192.436 84.503 0.750 re f 146.785 176.555 84.503 0.750 re f 146.785 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 152.035 182.581 Td /F1 9.8 Tf [(43.59#)] TJ ET 0.267 0.267 0.267 rg 230.538 192.436 71.899 0.750 re f 230.538 176.555 71.899 0.750 re f 230.538 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 235.788 182.581 Td /F1 9.8 Tf [(32.99#)] TJ ET 0.267 0.267 0.267 rg 301.687 192.436 70.566 0.750 re f 301.687 176.555 70.566 0.750 re f 301.687 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 306.937 182.581 Td /F1 9.8 Tf [(31.71#)] TJ ET 0.267 0.267 0.267 rg 371.503 192.436 71.899 0.750 re f 371.503 176.555 71.899 0.750 re f 371.503 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 376.753 182.581 Td /F1 9.8 Tf [(17.75#)] TJ ET 0.267 0.267 0.267 rg 442.651 192.436 63.486 0.750 re f 442.651 176.555 63.486 0.750 re f 442.651 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 447.901 182.581 Td /F1 9.8 Tf [(4.57*)] TJ ET 0.267 0.267 0.267 rg 505.387 192.436 75.488 0.750 re f 505.387 176.555 75.488 0.750 re f 505.387 176.555 0.750 16.631 re f 580.125 176.555 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.637 182.581 Td /F1 9.8 Tf [(NS)] TJ ET 0.965 0.965 0.965 rg 26.250 -334.570 555.000 465.750 re f 0.267 0.267 0.267 rg 26.250 131.180 m 581.250 131.180 l 581.250 130.430 l 26.250 130.430 l f q 278.250 0 0 450.000 35.250 -328.570 cm /I9 Do Q q 35.250 -334.570 537.000 0.000 re W n Q Q q 278.250 0 0 450.000 35.250 -328.570 cm /I9 Do Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(8)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Huntington Disease)] TJ ET Q endstream endobj 365 0 obj << /Type /Annot /Subtype /Link /A 366 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 -328.5698 313.5000 121.4302 ] >> endobj 366 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Figure-BACHD.jpg) >> endobj 367 0 obj << /Type /XObject /Subtype /Image /Width 371 /Height 600 /ColorSpace /DeviceRGB /Filter /DCTDecode /BitsPerComponent 8 /Length 53989>> stream JFIF;CREATOR: gd-jpeg v1.0 (using IJG JPEG v62), quality = 90 C     C   Xs" }!1AQa"q2#BR$3br %&'()*456789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz w!1AQaq"2B #3Rbr $4%&'()*56789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz ?Q4qK@ Fh▀E%⒀ EP3Gq@qK&h(qGfIQ-&hq@%/q@84f( ((8I@h-'f8 њ( 3Gq@qE((h? ? ^hh? ? ^hɠ(ϵ/44~~'GK~~'GK~~'KQ~~'GK'4~~'GK m蚷5by ≥cfGcOgڹO}n44:(e"Ƭ.pUo^(?6jZ&y$5Aw ONk~f j݂Agg4xTW$`(|yF| X$*IF?F;ְBh}cv)Bjzj'<7j:}zŒ*, .99Eu?* mwQ%߄t{:Q7Qʻ剉Bڹ =uͮ _\jW/-pp9$+u~ζ~ 'h3"eak F1r/>ܵZiyPH+f_;cq|I,/t5KE"Rv?=;Wgm3~_H:ھE$z cho{?zxD֧e}j֩t2:@988ǝ/k/i&MO실zx s= il`%z*7|?HfcxzmZ[|=RO8.2V#Aį|#BѬuxbo\[k5.kxcEHE 1Aq@~о _o}^ I4KW&F]W >*[O STW (wrci^ 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.[yqSV%`R~ #8v݉5'~y_kXڴPB_,sj5@2::7ŤWqu`+̾;^-/WرZv--Ϯ=G!p!*@gfs7hʺ:pH+~LKХƍqi(/loL$b?4K #`GWfRK>㵊)P,;wltux|A^jڬ7pm=2]|0\AzqQEwo,FC*tn`O=h=hqPwot3G+D$TpCR2ji(o2]i-O$ؙu2. 9 H}KZeZ&q%܍+>pv™oKXQz\Cmy̓?wn>|.3&N^9gXՎ{hF!7D ,2"Q/Ww0_NDPb2w`)Pi]*˹22ȡS+HV(cƽzs\b-f4|y;@lqJi.==hz2}袀hފ(ތ)h9^i1@MzQ@OzQ@OK&MPϽEϽPϽ/4Q'4sE/@ EϽ-%ϽѓE>sEsG>Q@>sKI@4sEsG>sG>uz9Ɗ((((?Z((((((((((((((Ɗ(((((( endstream endobj 368 0 obj << /Type /Annot /Subtype /Link /A 369 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 -328.5698 313.5000 121.4302 ] >> endobj 369 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Figure-BACHD.jpg) >> endobj 370 0 obj << /Type /Annot /Subtype /Link /A 371 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 -328.5698 313.5000 121.4302 ] >> endobj 371 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Figure-BACHD.jpg) >> endobj 372 0 obj << /Type /Page /Parent 3 0 R /Annots [ 374 0 R 377 0 R 379 0 R ] /Contents 373 0 R >> endobj 373 0 obj << /Length 36530 >> stream q 15.000 -174.113 577.500 951.113 re W n 0.965 0.965 0.965 rg 26.250 685.400 555.000 91.600 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 685.400 m 581.250 685.400 l 581.250 686.150 l 26.250 686.150 l f q 35.250 696.650 537.000 80.350 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 7: BACHD behavioral measures as a function of genotype, age, and light cycle.)] TJ ET BT 35.250 748.106 Td /F1 9.8 Tf [(*: significant genotype effect for each age within each cycle; **: significant genotype effect for each age, independent of )] TJ ET BT 35.250 734.370 Td /F1 9.8 Tf [(cycle; #: significant differences due to light cycle for each genotype and age; ##: significant differences due to light cycle for )] TJ ET BT 35.250 720.634 Td /F1 9.8 Tf [(each age independently of genotype; ###: significant differences due to light cycle independently of genotype and age; )] TJ ET BT 35.250 706.897 Td /F1 9.8 Tf [(####: significant differences due to light cycle and genotype independently of age.)] TJ ET Q q 26.250 664.164 555.000 13.736 re W n 0.271 0.267 0.267 rg BT 26.250 666.911 Td /F1 9.8 Tf [(Table 4. Summary of the statistical analyses for the computer vision derived measures for Experiment 2 \(BACHD\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 633.256 555.000 23.407 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 656.664 m 581.250 656.664 l 580.500 655.914 l 27.000 655.914 l f 581.250 656.664 m 581.250 633.256 l 580.500 633.256 l 580.500 655.914 l f 26.250 656.664 m 26.250 633.256 l 27.000 633.256 l 27.000 655.914 l f 0.271 0.267 0.267 rg BT 33.000 642.588 Td /F1 9.0 Tf [(Notes: *: p< 0.05, **: p< 0.01, #: p< 0.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 534.994 555.000 98.263 re f 0.965 0.965 0.965 rg 27.000 583.387 124.415 49.119 re f 0.267 0.267 0.267 rg 26.625 632.131 124.790 0.750 re f 26.625 620.227 0.750 12.655 re f 26.625 607.947 0.750 12.280 re f 26.625 595.667 0.750 12.280 re f 26.625 583.012 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 622.983 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 151.415 620.227 429.085 12.280 re f 0.267 0.267 0.267 rg 151.415 632.131 89.087 0.750 re f 240.502 632.131 66.731 0.750 re f 307.233 632.131 74.262 0.750 re f 381.495 632.131 66.731 0.750 re f 448.227 632.131 57.782 0.750 re f 506.009 632.131 74.866 0.750 re f 580.125 619.852 0.750 13.030 re f 0.271 0.267 0.267 rg BT 155.915 622.983 Td /F4 9.8 Tf [(BACHD)] TJ ET 0.965 0.965 0.965 rg 151.415 607.947 89.087 12.280 re f 0.267 0.267 0.267 rg 151.415 619.852 89.087 0.750 re f 0.271 0.267 0.267 rg BT 155.915 610.703 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 240.502 607.947 66.731 12.280 re f 0.267 0.267 0.267 rg 240.502 619.852 66.731 0.750 re f 0.271 0.267 0.267 rg BT 245.002 610.703 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 307.233 607.947 74.262 12.280 re f 0.267 0.267 0.267 rg 307.233 619.852 74.262 0.750 re f 0.271 0.267 0.267 rg BT 311.733 610.703 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 381.495 607.947 66.731 12.280 re f 0.267 0.267 0.267 rg 381.495 619.852 66.731 0.750 re f 0.271 0.267 0.267 rg BT 385.995 610.703 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 448.227 607.947 57.782 12.280 re f 0.267 0.267 0.267 rg 448.227 619.852 57.782 0.750 re f 0.271 0.267 0.267 rg BT 452.727 610.703 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 506.009 607.947 74.491 12.280 re f 0.267 0.267 0.267 rg 506.009 619.852 74.866 0.750 re f 580.125 607.572 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.509 610.703 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 151.415 595.667 89.087 12.280 re f 0.267 0.267 0.267 rg 151.415 607.572 89.087 0.750 re f 0.271 0.267 0.267 rg BT 155.915 598.423 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 240.502 595.667 66.731 12.280 re f 0.267 0.267 0.267 rg 240.502 607.572 66.731 0.750 re f 0.271 0.267 0.267 rg BT 245.002 598.423 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 307.233 595.667 74.262 12.280 re f 0.267 0.267 0.267 rg 307.233 607.572 74.262 0.750 re f 0.271 0.267 0.267 rg BT 311.733 598.423 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 381.495 595.667 66.731 12.280 re f 0.267 0.267 0.267 rg 381.495 607.572 66.731 0.750 re f 0.965 0.965 0.965 rg 448.227 595.667 57.782 12.280 re f 0.267 0.267 0.267 rg 448.227 607.572 57.782 0.750 re f 0.965 0.965 0.965 rg 506.009 595.667 74.491 12.280 re f 0.267 0.267 0.267 rg 506.009 607.572 74.866 0.750 re f 580.125 595.292 0.750 13.030 re f 0.965 0.965 0.965 rg 151.415 583.387 89.087 12.280 re f 0.267 0.267 0.267 rg 151.415 595.292 89.087 0.750 re f 0.271 0.267 0.267 rg BT 155.915 586.143 Td /F4 9.8 Tf [(F\(1,6\))] TJ ET 0.965 0.965 0.965 rg 240.502 583.387 66.731 12.280 re f 0.267 0.267 0.267 rg 240.502 595.292 66.731 0.750 re f 0.271 0.267 0.267 rg BT 245.002 586.143 Td /F4 9.8 Tf [(F\(3,18\))] TJ ET 0.965 0.965 0.965 rg 307.233 583.387 74.262 12.280 re f 0.267 0.267 0.267 rg 307.233 595.292 74.262 0.750 re f 0.271 0.267 0.267 rg BT 311.733 586.143 Td /F4 9.8 Tf [(F\(1,6\))] TJ ET 0.965 0.965 0.965 rg 381.495 583.387 66.731 12.280 re f 0.267 0.267 0.267 rg 381.495 595.292 66.731 0.750 re f 0.271 0.267 0.267 rg BT 385.995 586.143 Td /F4 9.8 Tf [(F\(3,18\))] TJ ET 0.965 0.965 0.965 rg 448.227 583.387 57.782 12.280 re f 0.267 0.267 0.267 rg 448.227 595.292 57.782 0.750 re f 0.271 0.267 0.267 rg BT 452.727 586.143 Td /F4 9.8 Tf [(F\(1,6\))] TJ ET 0.965 0.965 0.965 rg 506.009 583.387 74.491 12.280 re f 0.267 0.267 0.267 rg 506.009 595.292 74.866 0.750 re f 580.125 583.012 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.509 586.143 Td /F4 9.8 Tf [(F\(6,18\))] TJ ET 0.267 0.267 0.267 rg 26.625 583.012 125.165 0.750 re f 26.625 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 573.157 Td /F4 9.8 Tf [(Locomotion)] TJ ET 0.267 0.267 0.267 rg 151.040 583.012 89.837 0.750 re f 151.040 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 156.290 573.157 Td /F1 9.8 Tf [(149.53#)] TJ ET 0.267 0.267 0.267 rg 240.127 583.012 67.481 0.750 re f 240.127 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 245.377 573.157 Td /F1 9.8 Tf [(15.78#)] TJ ET 0.267 0.267 0.267 rg 306.858 583.012 75.012 0.750 re f 306.858 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 312.108 573.157 Td /F1 9.8 Tf [(127.02#)] TJ ET 0.267 0.267 0.267 rg 381.120 583.012 67.481 0.750 re f 381.120 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 386.370 573.157 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 447.852 583.012 58.532 0.750 re f 447.852 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 453.102 573.157 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.634 583.012 75.241 0.750 re f 505.634 567.131 0.750 16.631 re f 580.125 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.884 573.157 Td /F1 9.8 Tf [(9.79#)] TJ ET 0.267 0.267 0.267 rg 26.625 567.131 125.165 0.750 re f 26.625 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 557.276 Td /F4 9.8 Tf [(Immobility)] TJ ET 0.267 0.267 0.267 rg 151.040 567.131 89.837 0.750 re f 151.040 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 156.290 557.276 Td /F1 9.8 Tf [(58.09#)] TJ ET 0.267 0.267 0.267 rg 240.127 567.131 67.481 0.750 re f 240.127 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 245.377 557.276 Td /F1 9.8 Tf [(6.62**)] TJ ET 0.267 0.267 0.267 rg 306.858 567.131 75.012 0.750 re f 306.858 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 312.108 557.276 Td /F1 9.8 Tf [(12.88*)] TJ ET 0.267 0.267 0.267 rg 381.120 567.131 67.481 0.750 re f 381.120 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 386.370 557.276 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 447.852 567.131 58.532 0.750 re f 447.852 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 453.102 557.276 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.634 567.131 75.241 0.750 re f 505.634 551.250 0.750 16.631 re f 580.125 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.884 557.276 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 551.250 125.165 0.750 re f 26.625 535.369 125.165 0.750 re f 26.625 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 541.395 Td /F4 9.8 Tf [(Rearing Climbing)] TJ ET 0.267 0.267 0.267 rg 151.040 551.250 89.837 0.750 re f 151.040 535.369 89.837 0.750 re f 151.040 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 156.290 541.395 Td /F1 9.8 Tf [(7.22*)] TJ ET 0.267 0.267 0.267 rg 240.127 551.250 67.481 0.750 re f 240.127 535.369 67.481 0.750 re f 240.127 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 245.377 541.395 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 306.858 551.250 75.012 0.750 re f 306.858 535.369 75.012 0.750 re f 306.858 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 312.108 541.395 Td /F1 9.8 Tf [(18.82**)] TJ ET 0.267 0.267 0.267 rg 381.120 551.250 67.481 0.750 re f 381.120 535.369 67.481 0.750 re f 381.120 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 386.370 541.395 Td /F1 9.8 Tf [(4.05*)] TJ ET 0.267 0.267 0.267 rg 447.852 551.250 58.532 0.750 re f 447.852 535.369 58.532 0.750 re f 447.852 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 453.102 541.395 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.634 551.250 75.241 0.750 re f 505.634 535.369 75.241 0.750 re f 505.634 535.369 0.750 16.631 re f 580.125 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.884 541.395 Td /F1 9.8 Tf [(5.81**)] TJ ET BT 26.250 480.470 Td /F1 9.8 Tf [(Consistent with the R6/2 locomotor activity and motor function phenotype summarized above, BACHD mice exhibited lower )] TJ ET BT 26.250 468.565 Td /F1 9.8 Tf [(locomotor activity, higher immobility, and lower rearing and climbing than WT mice. Some of these differences were more )] TJ ET BT 26.250 456.660 Td /F1 9.8 Tf [(pronounced than in the R6/2 mice. Details of these results are summarized below.)] TJ ET BT 26.250 437.256 Td /F5 9.8 Tf [(Locomotion: )] TJ ET BT 82.069 437.256 Td /F1 9.8 Tf [(BACHD mice were hypoactive relative to WT control mice at all ages and during both cycle phases. Activity was )] TJ ET BT 26.250 425.351 Td /F1 9.8 Tf [(higher for both groups during the dark phase at 16, 24, and 36 weeks, as expected, but not at 52 weeks \(Figure 8A, Table 4; )] TJ ET BT 26.250 413.446 Td /F1 9.8 Tf [(Genotype x Age x Cycle: )] TJ ET BT 136.259 413.446 Td /F5 9.8 Tf [(p)] TJ ET BT 141.680 413.446 Td /F1 9.8 Tf [(< 0.001; simple main effects: )] TJ ET BT 268.762 413.446 Td /F5 9.8 Tf [(ps < )] TJ ET BT 290.173 413.446 Td /F1 9.8 Tf [(0)] TJ ET BT 295.594 413.446 Td /F5 9.8 Tf [(.)] TJ ET BT 298.304 413.446 Td /F1 9.8 Tf [(05\).)] TJ ET BT 26.250 394.041 Td /F5 9.8 Tf [(Time Immobile:)] TJ ET BT 92.882 394.041 Td /F1 9.8 Tf [( BACHD mice spent more time immobile than WT mice at all ages and light phases \(Figure 8B, Table 4, )] TJ ET BT 26.250 382.137 Td /F1 9.8 Tf [(Genotype main effect: )] TJ ET BT 124.345 382.137 Td /F5 9.8 Tf [(p)] TJ ET BT 129.766 382.137 Td /F1 9.8 Tf [( < 0.001\). Both groups spent more time immobile during the light phase and slightly more as they aged )] TJ ET BT 26.250 370.232 Td /F1 9.8 Tf [(\(Cycle and Age main effects: )] TJ ET BT 153.595 370.232 Td /F5 9.8 Tf [(p)] TJ ET BT 159.016 370.232 Td /F1 9.8 Tf [(s < 0.02\).)] TJ ET BT 26.250 350.827 Td /F5 9.8 Tf [(Rearing & Climbing:)] TJ ET BT 112.937 350.827 Td /F1 9.8 Tf [( BACHD mice reared and climbed less than WT mice at 16 and 52 weeks of age. WT reared and climbed )] TJ ET BT 26.250 338.922 Td /F1 9.8 Tf [(more during the dark at 16 and 36 ages but less at 52 weeks, whereas BACHD only climbed more at 36 weeks of age \(Figure )] TJ ET BT 26.250 327.018 Td /F1 9.8 Tf [(8C, Table 4, Genotype x Age x Cycle: )] TJ ET BT 192.078 327.018 Td /F5 9.8 Tf [(p )] TJ ET BT 200.209 327.018 Td /F1 9.8 Tf [(< 0.01; simple main effects: )] TJ ET BT 321.870 327.018 Td /F5 9.8 Tf [(p)] TJ ET BT 327.291 327.018 Td /F1 9.8 Tf [(s < .05)] TJ ET 0.965 0.965 0.965 rg 26.250 -174.113 555.000 491.250 re f 0.267 0.267 0.267 rg 26.250 317.137 m 581.250 317.137 l 581.250 316.387 l 26.250 316.387 l f q 450.000 0 0 475.500 35.250 -168.113 cm /I10 Do Q q 35.250 -174.113 537.000 0.000 re W n Q Q q 15.000 -174.113 577.500 951.113 re W n 0.965 0.965 0.965 rg 26.250 685.400 555.000 91.600 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 685.400 m 581.250 685.400 l 581.250 686.150 l 26.250 686.150 l f q 35.250 696.650 537.000 80.350 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 7: BACHD behavioral measures as a function of genotype, age, and light cycle.)] TJ ET BT 35.250 748.106 Td /F1 9.8 Tf [(*: significant genotype effect for each age within each cycle; **: significant genotype effect for each age, independent of )] TJ ET BT 35.250 734.370 Td /F1 9.8 Tf [(cycle; #: significant differences due to light cycle for each genotype and age; ##: significant differences due to light cycle for )] TJ ET BT 35.250 720.634 Td /F1 9.8 Tf [(each age independently of genotype; ###: significant differences due to light cycle independently of genotype and age; )] TJ ET BT 35.250 706.897 Td /F1 9.8 Tf [(####: significant differences due to light cycle and genotype independently of age.)] TJ ET Q q 26.250 664.164 555.000 13.736 re W n 0.271 0.267 0.267 rg BT 26.250 666.911 Td /F1 9.8 Tf [(Table 4. Summary of the statistical analyses for the computer vision derived measures for Experiment 2 \(BACHD\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 633.256 555.000 23.407 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 656.664 m 581.250 656.664 l 580.500 655.914 l 27.000 655.914 l f 581.250 656.664 m 581.250 633.256 l 580.500 633.256 l 580.500 655.914 l f 26.250 656.664 m 26.250 633.256 l 27.000 633.256 l 27.000 655.914 l f 0.271 0.267 0.267 rg BT 33.000 642.588 Td /F1 9.0 Tf [(Notes: *: p< 0.05, **: p< 0.01, #: p< 0.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 534.994 555.000 98.263 re f 0.965 0.965 0.965 rg 27.000 583.387 124.415 49.119 re f 0.267 0.267 0.267 rg 26.625 632.131 124.790 0.750 re f 26.625 620.227 0.750 12.655 re f 26.625 607.947 0.750 12.280 re f 26.625 595.667 0.750 12.280 re f 26.625 583.012 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 622.983 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 151.415 620.227 429.085 12.280 re f 0.267 0.267 0.267 rg 151.415 632.131 89.087 0.750 re f 240.502 632.131 66.731 0.750 re f 307.233 632.131 74.262 0.750 re f 381.495 632.131 66.731 0.750 re f 448.227 632.131 57.782 0.750 re f 506.009 632.131 74.866 0.750 re f 580.125 619.852 0.750 13.030 re f 0.271 0.267 0.267 rg BT 155.915 622.983 Td /F4 9.8 Tf [(BACHD)] TJ ET 0.965 0.965 0.965 rg 151.415 607.947 89.087 12.280 re f 0.267 0.267 0.267 rg 151.415 619.852 89.087 0.750 re f 0.271 0.267 0.267 rg BT 155.915 610.703 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 240.502 607.947 66.731 12.280 re f 0.267 0.267 0.267 rg 240.502 619.852 66.731 0.750 re f 0.271 0.267 0.267 rg BT 245.002 610.703 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 307.233 607.947 74.262 12.280 re f 0.267 0.267 0.267 rg 307.233 619.852 74.262 0.750 re f 0.271 0.267 0.267 rg BT 311.733 610.703 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 381.495 607.947 66.731 12.280 re f 0.267 0.267 0.267 rg 381.495 619.852 66.731 0.750 re f 0.271 0.267 0.267 rg BT 385.995 610.703 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 448.227 607.947 57.782 12.280 re f 0.267 0.267 0.267 rg 448.227 619.852 57.782 0.750 re f 0.271 0.267 0.267 rg BT 452.727 610.703 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 506.009 607.947 74.491 12.280 re f 0.267 0.267 0.267 rg 506.009 619.852 74.866 0.750 re f 580.125 607.572 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.509 610.703 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 151.415 595.667 89.087 12.280 re f 0.267 0.267 0.267 rg 151.415 607.572 89.087 0.750 re f 0.271 0.267 0.267 rg BT 155.915 598.423 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 240.502 595.667 66.731 12.280 re f 0.267 0.267 0.267 rg 240.502 607.572 66.731 0.750 re f 0.271 0.267 0.267 rg BT 245.002 598.423 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 307.233 595.667 74.262 12.280 re f 0.267 0.267 0.267 rg 307.233 607.572 74.262 0.750 re f 0.271 0.267 0.267 rg BT 311.733 598.423 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 381.495 595.667 66.731 12.280 re f 0.267 0.267 0.267 rg 381.495 607.572 66.731 0.750 re f 0.965 0.965 0.965 rg 448.227 595.667 57.782 12.280 re f 0.267 0.267 0.267 rg 448.227 607.572 57.782 0.750 re f 0.965 0.965 0.965 rg 506.009 595.667 74.491 12.280 re f 0.267 0.267 0.267 rg 506.009 607.572 74.866 0.750 re f 580.125 595.292 0.750 13.030 re f 0.965 0.965 0.965 rg 151.415 583.387 89.087 12.280 re f 0.267 0.267 0.267 rg 151.415 595.292 89.087 0.750 re f 0.271 0.267 0.267 rg BT 155.915 586.143 Td /F4 9.8 Tf [(F\(1,6\))] TJ ET 0.965 0.965 0.965 rg 240.502 583.387 66.731 12.280 re f 0.267 0.267 0.267 rg 240.502 595.292 66.731 0.750 re f 0.271 0.267 0.267 rg BT 245.002 586.143 Td /F4 9.8 Tf [(F\(3,18\))] TJ ET 0.965 0.965 0.965 rg 307.233 583.387 74.262 12.280 re f 0.267 0.267 0.267 rg 307.233 595.292 74.262 0.750 re f 0.271 0.267 0.267 rg BT 311.733 586.143 Td /F4 9.8 Tf [(F\(1,6\))] TJ ET 0.965 0.965 0.965 rg 381.495 583.387 66.731 12.280 re f 0.267 0.267 0.267 rg 381.495 595.292 66.731 0.750 re f 0.271 0.267 0.267 rg BT 385.995 586.143 Td /F4 9.8 Tf [(F\(3,18\))] TJ ET 0.965 0.965 0.965 rg 448.227 583.387 57.782 12.280 re f 0.267 0.267 0.267 rg 448.227 595.292 57.782 0.750 re f 0.271 0.267 0.267 rg BT 452.727 586.143 Td /F4 9.8 Tf [(F\(1,6\))] TJ ET 0.965 0.965 0.965 rg 506.009 583.387 74.491 12.280 re f 0.267 0.267 0.267 rg 506.009 595.292 74.866 0.750 re f 580.125 583.012 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.509 586.143 Td /F4 9.8 Tf [(F\(6,18\))] TJ ET 0.267 0.267 0.267 rg 26.625 583.012 125.165 0.750 re f 26.625 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 573.157 Td /F4 9.8 Tf [(Locomotion)] TJ ET 0.267 0.267 0.267 rg 151.040 583.012 89.837 0.750 re f 151.040 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 156.290 573.157 Td /F1 9.8 Tf [(149.53#)] TJ ET 0.267 0.267 0.267 rg 240.127 583.012 67.481 0.750 re f 240.127 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 245.377 573.157 Td /F1 9.8 Tf [(15.78#)] TJ ET 0.267 0.267 0.267 rg 306.858 583.012 75.012 0.750 re f 306.858 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 312.108 573.157 Td /F1 9.8 Tf [(127.02#)] TJ ET 0.267 0.267 0.267 rg 381.120 583.012 67.481 0.750 re f 381.120 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 386.370 573.157 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 447.852 583.012 58.532 0.750 re f 447.852 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 453.102 573.157 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.634 583.012 75.241 0.750 re f 505.634 567.131 0.750 16.631 re f 580.125 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.884 573.157 Td /F1 9.8 Tf [(9.79#)] TJ ET 0.267 0.267 0.267 rg 26.625 567.131 125.165 0.750 re f 26.625 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 557.276 Td /F4 9.8 Tf [(Immobility)] TJ ET 0.267 0.267 0.267 rg 151.040 567.131 89.837 0.750 re f 151.040 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 156.290 557.276 Td /F1 9.8 Tf [(58.09#)] TJ ET 0.267 0.267 0.267 rg 240.127 567.131 67.481 0.750 re f 240.127 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 245.377 557.276 Td /F1 9.8 Tf [(6.62**)] TJ ET 0.267 0.267 0.267 rg 306.858 567.131 75.012 0.750 re f 306.858 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 312.108 557.276 Td /F1 9.8 Tf [(12.88*)] TJ ET 0.267 0.267 0.267 rg 381.120 567.131 67.481 0.750 re f 381.120 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 386.370 557.276 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 447.852 567.131 58.532 0.750 re f 447.852 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 453.102 557.276 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.634 567.131 75.241 0.750 re f 505.634 551.250 0.750 16.631 re f 580.125 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.884 557.276 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 551.250 125.165 0.750 re f 26.625 535.369 125.165 0.750 re f 26.625 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 541.395 Td /F4 9.8 Tf [(Rearing Climbing)] TJ ET 0.267 0.267 0.267 rg 151.040 551.250 89.837 0.750 re f 151.040 535.369 89.837 0.750 re f 151.040 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 156.290 541.395 Td /F1 9.8 Tf [(7.22*)] TJ ET 0.267 0.267 0.267 rg 240.127 551.250 67.481 0.750 re f 240.127 535.369 67.481 0.750 re f 240.127 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 245.377 541.395 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 306.858 551.250 75.012 0.750 re f 306.858 535.369 75.012 0.750 re f 306.858 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 312.108 541.395 Td /F1 9.8 Tf [(18.82**)] TJ ET 0.267 0.267 0.267 rg 381.120 551.250 67.481 0.750 re f 381.120 535.369 67.481 0.750 re f 381.120 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 386.370 541.395 Td /F1 9.8 Tf [(4.05*)] TJ ET 0.267 0.267 0.267 rg 447.852 551.250 58.532 0.750 re f 447.852 535.369 58.532 0.750 re f 447.852 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 453.102 541.395 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.634 551.250 75.241 0.750 re f 505.634 535.369 75.241 0.750 re f 505.634 535.369 0.750 16.631 re f 580.125 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.884 541.395 Td /F1 9.8 Tf [(5.81**)] TJ ET BT 26.250 480.470 Td /F1 9.8 Tf [(Consistent with the R6/2 locomotor activity and motor function phenotype summarized above, BACHD mice exhibited lower )] TJ ET BT 26.250 468.565 Td /F1 9.8 Tf [(locomotor activity, higher immobility, and lower rearing and climbing than WT mice. Some of these differences were more )] TJ ET BT 26.250 456.660 Td /F1 9.8 Tf [(pronounced than in the R6/2 mice. Details of these results are summarized below.)] TJ ET BT 26.250 437.256 Td /F5 9.8 Tf [(Locomotion: )] TJ ET BT 82.069 437.256 Td /F1 9.8 Tf [(BACHD mice were hypoactive relative to WT control mice at all ages and during both cycle phases. Activity was )] TJ ET BT 26.250 425.351 Td /F1 9.8 Tf [(higher for both groups during the dark phase at 16, 24, and 36 weeks, as expected, but not at 52 weeks \(Figure 8A, Table 4; )] TJ ET BT 26.250 413.446 Td /F1 9.8 Tf [(Genotype x Age x Cycle: )] TJ ET BT 136.259 413.446 Td /F5 9.8 Tf [(p)] TJ ET BT 141.680 413.446 Td /F1 9.8 Tf [(< 0.001; simple main effects: )] TJ ET BT 268.762 413.446 Td /F5 9.8 Tf [(ps < )] TJ ET BT 290.173 413.446 Td /F1 9.8 Tf [(0)] TJ ET BT 295.594 413.446 Td /F5 9.8 Tf [(.)] TJ ET BT 298.304 413.446 Td /F1 9.8 Tf [(05\).)] TJ ET BT 26.250 394.041 Td /F5 9.8 Tf [(Time Immobile:)] TJ ET BT 92.882 394.041 Td /F1 9.8 Tf [( BACHD mice spent more time immobile than WT mice at all ages and light phases \(Figure 8B, Table 4, )] TJ ET BT 26.250 382.137 Td /F1 9.8 Tf [(Genotype main effect: )] TJ ET BT 124.345 382.137 Td /F5 9.8 Tf [(p)] TJ ET BT 129.766 382.137 Td /F1 9.8 Tf [( < 0.001\). Both groups spent more time immobile during the light phase and slightly more as they aged )] TJ ET BT 26.250 370.232 Td /F1 9.8 Tf [(\(Cycle and Age main effects: )] TJ ET BT 153.595 370.232 Td /F5 9.8 Tf [(p)] TJ ET BT 159.016 370.232 Td /F1 9.8 Tf [(s < 0.02\).)] TJ ET BT 26.250 350.827 Td /F5 9.8 Tf [(Rearing & Climbing:)] TJ ET BT 112.937 350.827 Td /F1 9.8 Tf [( BACHD mice reared and climbed less than WT mice at 16 and 52 weeks of age. WT reared and climbed )] TJ ET BT 26.250 338.922 Td /F1 9.8 Tf [(more during the dark at 16 and 36 ages but less at 52 weeks, whereas BACHD only climbed more at 36 weeks of age \(Figure )] TJ ET BT 26.250 327.018 Td /F1 9.8 Tf [(8C, Table 4, Genotype x Age x Cycle: )] TJ ET BT 192.078 327.018 Td /F5 9.8 Tf [(p )] TJ ET BT 200.209 327.018 Td /F1 9.8 Tf [(< 0.01; simple main effects: )] TJ ET BT 321.870 327.018 Td /F5 9.8 Tf [(p)] TJ ET BT 327.291 327.018 Td /F1 9.8 Tf [(s < .05)] TJ ET 0.965 0.965 0.965 rg 26.250 -174.113 555.000 491.250 re f 0.267 0.267 0.267 rg 26.250 317.137 m 581.250 317.137 l 581.250 316.387 l 26.250 316.387 l f q 450.000 0 0 475.500 35.250 -168.113 cm /I10 Do Q q 35.250 -174.113 537.000 0.000 re W n Q Q q 15.000 -174.113 577.500 951.113 re W n 0.965 0.965 0.965 rg 26.250 685.400 555.000 91.600 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 685.400 m 581.250 685.400 l 581.250 686.150 l 26.250 686.150 l f q 35.250 696.650 537.000 80.350 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 7: BACHD behavioral measures as a function of genotype, age, and light cycle.)] TJ ET BT 35.250 748.106 Td /F1 9.8 Tf [(*: significant genotype effect for each age within each cycle; **: significant genotype effect for each age, independent of )] TJ ET BT 35.250 734.370 Td /F1 9.8 Tf [(cycle; #: significant differences due to light cycle for each genotype and age; ##: significant differences due to light cycle for )] TJ ET BT 35.250 720.634 Td /F1 9.8 Tf [(each age independently of genotype; ###: significant differences due to light cycle independently of genotype and age; )] TJ ET BT 35.250 706.897 Td /F1 9.8 Tf [(####: significant differences due to light cycle and genotype independently of age.)] TJ ET Q q 26.250 664.164 555.000 13.736 re W n 0.271 0.267 0.267 rg BT 26.250 666.911 Td /F1 9.8 Tf [(Table 4. Summary of the statistical analyses for the computer vision derived measures for Experiment 2 \(BACHD\).)] TJ ET Q 0.965 0.965 0.965 rg 26.250 633.256 555.000 23.407 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 656.664 m 581.250 656.664 l 580.500 655.914 l 27.000 655.914 l f 581.250 656.664 m 581.250 633.256 l 580.500 633.256 l 580.500 655.914 l f 26.250 656.664 m 26.250 633.256 l 27.000 633.256 l 27.000 655.914 l f 0.271 0.267 0.267 rg BT 33.000 642.588 Td /F1 9.0 Tf [(Notes: *: p< 0.05, **: p< 0.01, #: p< 0.001, NS: p>.05)] TJ ET 1.000 1.000 1.000 rg 26.250 534.994 555.000 98.263 re f 0.965 0.965 0.965 rg 27.000 583.387 124.415 49.119 re f 0.267 0.267 0.267 rg 26.625 632.131 124.790 0.750 re f 26.625 620.227 0.750 12.655 re f 26.625 607.947 0.750 12.280 re f 26.625 595.667 0.750 12.280 re f 26.625 583.012 0.750 12.655 re f 0.271 0.267 0.267 rg BT 31.500 622.983 Td /F4 9.8 Tf [(Behavior)] TJ ET 0.965 0.965 0.965 rg 151.415 620.227 429.085 12.280 re f 0.267 0.267 0.267 rg 151.415 632.131 89.087 0.750 re f 240.502 632.131 66.731 0.750 re f 307.233 632.131 74.262 0.750 re f 381.495 632.131 66.731 0.750 re f 448.227 632.131 57.782 0.750 re f 506.009 632.131 74.866 0.750 re f 580.125 619.852 0.750 13.030 re f 0.271 0.267 0.267 rg BT 155.915 622.983 Td /F4 9.8 Tf [(BACHD)] TJ ET 0.965 0.965 0.965 rg 151.415 607.947 89.087 12.280 re f 0.267 0.267 0.267 rg 151.415 619.852 89.087 0.750 re f 0.271 0.267 0.267 rg BT 155.915 610.703 Td /F4 9.8 Tf [(Genotype)] TJ ET 0.965 0.965 0.965 rg 240.502 607.947 66.731 12.280 re f 0.267 0.267 0.267 rg 240.502 619.852 66.731 0.750 re f 0.271 0.267 0.267 rg BT 245.002 610.703 Td /F4 9.8 Tf [(Age)] TJ ET 0.965 0.965 0.965 rg 307.233 607.947 74.262 12.280 re f 0.267 0.267 0.267 rg 307.233 619.852 74.262 0.750 re f 0.271 0.267 0.267 rg BT 311.733 610.703 Td /F4 9.8 Tf [(Cycle)] TJ ET 0.965 0.965 0.965 rg 381.495 607.947 66.731 12.280 re f 0.267 0.267 0.267 rg 381.495 619.852 66.731 0.750 re f 0.271 0.267 0.267 rg BT 385.995 610.703 Td /F4 9.8 Tf [(G x A)] TJ ET 0.965 0.965 0.965 rg 448.227 607.947 57.782 12.280 re f 0.267 0.267 0.267 rg 448.227 619.852 57.782 0.750 re f 0.271 0.267 0.267 rg BT 452.727 610.703 Td /F4 9.8 Tf [(G x C)] TJ ET 0.965 0.965 0.965 rg 506.009 607.947 74.491 12.280 re f 0.267 0.267 0.267 rg 506.009 619.852 74.866 0.750 re f 580.125 607.572 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.509 610.703 Td /F4 9.8 Tf [(G x A x C)] TJ ET 0.965 0.965 0.965 rg 151.415 595.667 89.087 12.280 re f 0.267 0.267 0.267 rg 151.415 607.572 89.087 0.750 re f 0.271 0.267 0.267 rg BT 155.915 598.423 Td /F4 9.8 Tf [(\(G\))] TJ ET 0.965 0.965 0.965 rg 240.502 595.667 66.731 12.280 re f 0.267 0.267 0.267 rg 240.502 607.572 66.731 0.750 re f 0.271 0.267 0.267 rg BT 245.002 598.423 Td /F4 9.8 Tf [(\(A\))] TJ ET 0.965 0.965 0.965 rg 307.233 595.667 74.262 12.280 re f 0.267 0.267 0.267 rg 307.233 607.572 74.262 0.750 re f 0.271 0.267 0.267 rg BT 311.733 598.423 Td /F4 9.8 Tf [(\(C\))] TJ ET 0.965 0.965 0.965 rg 381.495 595.667 66.731 12.280 re f 0.267 0.267 0.267 rg 381.495 607.572 66.731 0.750 re f 0.965 0.965 0.965 rg 448.227 595.667 57.782 12.280 re f 0.267 0.267 0.267 rg 448.227 607.572 57.782 0.750 re f 0.965 0.965 0.965 rg 506.009 595.667 74.491 12.280 re f 0.267 0.267 0.267 rg 506.009 607.572 74.866 0.750 re f 580.125 595.292 0.750 13.030 re f 0.965 0.965 0.965 rg 151.415 583.387 89.087 12.280 re f 0.267 0.267 0.267 rg 151.415 595.292 89.087 0.750 re f 0.271 0.267 0.267 rg BT 155.915 586.143 Td /F4 9.8 Tf [(F\(1,6\))] TJ ET 0.965 0.965 0.965 rg 240.502 583.387 66.731 12.280 re f 0.267 0.267 0.267 rg 240.502 595.292 66.731 0.750 re f 0.271 0.267 0.267 rg BT 245.002 586.143 Td /F4 9.8 Tf [(F\(3,18\))] TJ ET 0.965 0.965 0.965 rg 307.233 583.387 74.262 12.280 re f 0.267 0.267 0.267 rg 307.233 595.292 74.262 0.750 re f 0.271 0.267 0.267 rg BT 311.733 586.143 Td /F4 9.8 Tf [(F\(1,6\))] TJ ET 0.965 0.965 0.965 rg 381.495 583.387 66.731 12.280 re f 0.267 0.267 0.267 rg 381.495 595.292 66.731 0.750 re f 0.271 0.267 0.267 rg BT 385.995 586.143 Td /F4 9.8 Tf [(F\(3,18\))] TJ ET 0.965 0.965 0.965 rg 448.227 583.387 57.782 12.280 re f 0.267 0.267 0.267 rg 448.227 595.292 57.782 0.750 re f 0.271 0.267 0.267 rg BT 452.727 586.143 Td /F4 9.8 Tf [(F\(1,6\))] TJ ET 0.965 0.965 0.965 rg 506.009 583.387 74.491 12.280 re f 0.267 0.267 0.267 rg 506.009 595.292 74.866 0.750 re f 580.125 583.012 0.750 13.030 re f 0.271 0.267 0.267 rg BT 510.509 586.143 Td /F4 9.8 Tf [(F\(6,18\))] TJ ET 0.267 0.267 0.267 rg 26.625 583.012 125.165 0.750 re f 26.625 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 573.157 Td /F4 9.8 Tf [(Locomotion)] TJ ET 0.267 0.267 0.267 rg 151.040 583.012 89.837 0.750 re f 151.040 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 156.290 573.157 Td /F1 9.8 Tf [(149.53#)] TJ ET 0.267 0.267 0.267 rg 240.127 583.012 67.481 0.750 re f 240.127 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 245.377 573.157 Td /F1 9.8 Tf [(15.78#)] TJ ET 0.267 0.267 0.267 rg 306.858 583.012 75.012 0.750 re f 306.858 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 312.108 573.157 Td /F1 9.8 Tf [(127.02#)] TJ ET 0.267 0.267 0.267 rg 381.120 583.012 67.481 0.750 re f 381.120 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 386.370 573.157 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 447.852 583.012 58.532 0.750 re f 447.852 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 453.102 573.157 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.634 583.012 75.241 0.750 re f 505.634 567.131 0.750 16.631 re f 580.125 567.131 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.884 573.157 Td /F1 9.8 Tf [(9.79#)] TJ ET 0.267 0.267 0.267 rg 26.625 567.131 125.165 0.750 re f 26.625 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 557.276 Td /F4 9.8 Tf [(Immobility)] TJ ET 0.267 0.267 0.267 rg 151.040 567.131 89.837 0.750 re f 151.040 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 156.290 557.276 Td /F1 9.8 Tf [(58.09#)] TJ ET 0.267 0.267 0.267 rg 240.127 567.131 67.481 0.750 re f 240.127 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 245.377 557.276 Td /F1 9.8 Tf [(6.62**)] TJ ET 0.267 0.267 0.267 rg 306.858 567.131 75.012 0.750 re f 306.858 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 312.108 557.276 Td /F1 9.8 Tf [(12.88*)] TJ ET 0.267 0.267 0.267 rg 381.120 567.131 67.481 0.750 re f 381.120 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 386.370 557.276 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 447.852 567.131 58.532 0.750 re f 447.852 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 453.102 557.276 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.634 567.131 75.241 0.750 re f 505.634 551.250 0.750 16.631 re f 580.125 551.250 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.884 557.276 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 26.625 551.250 125.165 0.750 re f 26.625 535.369 125.165 0.750 re f 26.625 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 31.875 541.395 Td /F4 9.8 Tf [(Rearing Climbing)] TJ ET 0.267 0.267 0.267 rg 151.040 551.250 89.837 0.750 re f 151.040 535.369 89.837 0.750 re f 151.040 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 156.290 541.395 Td /F1 9.8 Tf [(7.22*)] TJ ET 0.267 0.267 0.267 rg 240.127 551.250 67.481 0.750 re f 240.127 535.369 67.481 0.750 re f 240.127 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 245.377 541.395 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 306.858 551.250 75.012 0.750 re f 306.858 535.369 75.012 0.750 re f 306.858 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 312.108 541.395 Td /F1 9.8 Tf [(18.82**)] TJ ET 0.267 0.267 0.267 rg 381.120 551.250 67.481 0.750 re f 381.120 535.369 67.481 0.750 re f 381.120 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 386.370 541.395 Td /F1 9.8 Tf [(4.05*)] TJ ET 0.267 0.267 0.267 rg 447.852 551.250 58.532 0.750 re f 447.852 535.369 58.532 0.750 re f 447.852 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 453.102 541.395 Td /F1 9.8 Tf [(NS)] TJ ET 0.267 0.267 0.267 rg 505.634 551.250 75.241 0.750 re f 505.634 535.369 75.241 0.750 re f 505.634 535.369 0.750 16.631 re f 580.125 535.369 0.750 16.631 re f 0.271 0.267 0.267 rg BT 510.884 541.395 Td /F1 9.8 Tf [(5.81**)] TJ ET BT 26.250 480.470 Td /F1 9.8 Tf [(Consistent with the R6/2 locomotor activity and motor function phenotype summarized above, BACHD mice exhibited lower )] TJ ET BT 26.250 468.565 Td /F1 9.8 Tf [(locomotor activity, higher immobility, and lower rearing and climbing than WT mice. Some of these differences were more )] TJ ET BT 26.250 456.660 Td /F1 9.8 Tf [(pronounced than in the R6/2 mice. Details of these results are summarized below.)] TJ ET BT 26.250 437.256 Td /F5 9.8 Tf [(Locomotion: )] TJ ET BT 82.069 437.256 Td /F1 9.8 Tf [(BACHD mice were hypoactive relative to WT control mice at all ages and during both cycle phases. Activity was )] TJ ET BT 26.250 425.351 Td /F1 9.8 Tf [(higher for both groups during the dark phase at 16, 24, and 36 weeks, as expected, but not at 52 weeks \(Figure 8A, Table 4; )] TJ ET BT 26.250 413.446 Td /F1 9.8 Tf [(Genotype x Age x Cycle: )] TJ ET BT 136.259 413.446 Td /F5 9.8 Tf [(p)] TJ ET BT 141.680 413.446 Td /F1 9.8 Tf [(< 0.001; simple main effects: )] TJ ET BT 268.762 413.446 Td /F5 9.8 Tf [(ps < )] TJ ET BT 290.173 413.446 Td /F1 9.8 Tf [(0)] TJ ET BT 295.594 413.446 Td /F5 9.8 Tf [(.)] TJ ET BT 298.304 413.446 Td /F1 9.8 Tf [(05\).)] TJ ET BT 26.250 394.041 Td /F5 9.8 Tf [(Time Immobile:)] TJ ET BT 92.882 394.041 Td /F1 9.8 Tf [( BACHD mice spent more time immobile than WT mice at all ages and light phases \(Figure 8B, Table 4, )] TJ ET BT 26.250 382.137 Td /F1 9.8 Tf [(Genotype main effect: )] TJ ET BT 124.345 382.137 Td /F5 9.8 Tf [(p)] TJ ET BT 129.766 382.137 Td /F1 9.8 Tf [( < 0.001\). Both groups spent more time immobile during the light phase and slightly more as they aged )] TJ ET BT 26.250 370.232 Td /F1 9.8 Tf [(\(Cycle and Age main effects: )] TJ ET BT 153.595 370.232 Td /F5 9.8 Tf [(p)] TJ ET BT 159.016 370.232 Td /F1 9.8 Tf [(s < 0.02\).)] TJ ET BT 26.250 350.827 Td /F5 9.8 Tf [(Rearing & Climbing:)] TJ ET BT 112.937 350.827 Td /F1 9.8 Tf [( BACHD mice reared and climbed less than WT mice at 16 and 52 weeks of age. WT reared and climbed )] TJ ET BT 26.250 338.922 Td /F1 9.8 Tf [(more during the dark at 16 and 36 ages but less at 52 weeks, whereas BACHD only climbed more at 36 weeks of age \(Figure )] TJ ET BT 26.250 327.018 Td /F1 9.8 Tf [(8C, Table 4, Genotype x Age x Cycle: )] TJ ET BT 192.078 327.018 Td /F5 9.8 Tf [(p )] TJ ET BT 200.209 327.018 Td /F1 9.8 Tf [(< 0.01; simple main effects: )] TJ ET BT 321.870 327.018 Td /F5 9.8 Tf [(p)] TJ ET BT 327.291 327.018 Td /F1 9.8 Tf [(s < .05)] TJ ET 0.965 0.965 0.965 rg 26.250 -174.113 555.000 491.250 re f 0.267 0.267 0.267 rg 26.250 317.137 m 581.250 317.137 l 581.250 316.387 l 26.250 316.387 l f q 450.000 0 0 475.500 35.250 -168.113 cm /I10 Do Q q 35.250 -174.113 537.000 0.000 re W n Q Q q 450.000 0 0 475.500 35.250 -168.113 cm /I10 Do Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(9)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Huntington Disease)] TJ ET Q endstream endobj 374 0 obj << /Type /Annot /Subtype /Link /A 375 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 -168.1133 485.2500 307.3867 ] >> endobj 375 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Figure-BACHD-A1.jpg) >> endobj 376 0 obj << /Type /XObject /Subtype /Image /Width 600 /Height 634 /ColorSpace /DeviceRGB /Filter /DCTDecode /BitsPerComponent 8 /Length 67621>> stream JFIF;CREATOR: gd-jpeg v1.0 (using IJG JPEG v62), quality = 90 C     C   zX" }!1AQa"q2#BR$3br %&'()*456789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz w!1AQaq"2B #3Rbr $4%&'()*56789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz ?QbPތъ1@Q3@b֎bP(gJ(<њ1F(@b (?ʊ1EfQ@hb hfъ1@ I(.i3F(b ;ъ1@isI1@z3F(4Q(Fh4PhZ1F(}XY\ʹH S Ǥt_UnCKo:vi~#βmL'S^3~_QtNu{9!"AN9:P(nZ-H+BsQXII#u7Pr=>Rb_|ux#[Eh)܉n1U*@"b?j5i}3FC AW~42^F[HHYzCBpq3@(Oo |\ҵ Gzvi2n]D  8qG+x[4DŽmU,Af431.0\qF+˾1ӟ _i6~:ZHpiUL Fnk[▽Ȭ.ĖJވ*>444f Rbb3EQ1@%.(%-RRPIK1@ E.(Q1@c1@ KF1F(1@bPIK1@%.(%-b J\Q))qF(%-bPRъ1@ KF(|}X/![;vH2ڄ!pA ~' c n#G`2>0?e^|{xχ}jeH8&ym?!^ydOx9r4OP__?o\:gmKR/?|a[j7'N4+8VR%3^W>'ͩ3O/rT+kX#BG (G@ھ/pG@(# q(|t!|7<q7nged_p'eS|dӕ&xfMeaeH G+|{iDVwÚvme>_ $|̴dl9?<B9u .FtI/"%T0b;yPy"/P->!ɬZbi4fbCÿ|)м7^!7wIz_ +zw6~ EN.Hːm wA?Jx#/ƺ|'[O]m0RB o'gPÿ?W> xw*]XjBݭ'1G%O~ձOo~͇^ ]ti:^lo~|Cy?="V8ޝȧ~u,}:*0Fp(4 6+?[ʰs^U~ 6Zimg-ޛC ͕)dtnQ^5SD79>"լM"OD,0Ǻ؀2H5п/Z zi,4}n/yRcʪ.w~?~vb]>ZvF;UT[πض/L+=۞k1@ KF(Q(-((h((hFy((sE'G@ (-/z)? ? ^Q'@ E'F}h((h)s@~-f€ (ϵ'@ E&}(h.}ŒPE(hϵ-~@ xŵ ybIFѕІRA T?~ϵP7_Fݭ4rĚ>8ei# pkHXcHmE@Qjs_|=.Z<=2m#=EgiVpm+omimHFGU—>W/-k^Z֜ɨZs]bebO=k((Ÿ5iy=XhM}~P,Fv,3S]i? ? 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I':'ӡh_~ F +BvL."} X0 Q\+ н$V<%`|3"0+2@4RQ@ MPQ(4P1FhQFhQ1F(bzQڀ Rdf~:1CPl>+b伵>nWG̈́7ce^Kvv(x"m;q/?8q8h.r881潎v4 NdWn%x_$xo\܁/ٟ%22 ǡoXɪh;D…B:s JæYgM%QI3ID=6{o㲷OR/ogzE_z>(T1F(EE%((EQE襤J(((?*((aifH%cρZ((~+[mGLO6Kt~>d,zVP(((( Az((E"eP riPGz(QE-'C@;QEPEPE-% U'RRTR@ A?*ZLQKI@AE-%-%-%~4Pt}h%/E%Q@ @-hғhhi-hv4Q@@z3IڏO)JJZ( N@'RRQ@@QEPh(4Ph֊(@Ph4P: (Ҁ;h(ŽQ@ƎQAEv4)h=EQޏZ3KIڒZ-%( endstream endobj 377 0 obj << /Type /Annot /Subtype /Link /A 378 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 -168.1133 485.2500 307.3867 ] >> endobj 378 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Figure-BACHD-A1.jpg) >> endobj 379 0 obj << /Type /Annot /Subtype /Link /A 380 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 -168.1133 485.2500 307.3867 ] >> endobj 380 0 obj << /Type /Action /S /URI /URI (https://currents.plos.org/hd/files/2012/11/Figure-BACHD-A1.jpg) >> endobj 381 0 obj << /Type /Page /Parent 3 0 R /Annots [ 383 0 R 385 0 R 387 0 R 389 0 R 391 0 R 393 0 R 395 0 R 397 0 R 399 0 R 401 0 R 403 0 R 405 0 R 407 0 R 409 0 R 411 0 R 413 0 R 415 0 R 417 0 R 419 0 R 421 0 R 423 0 R 425 0 R 427 0 R 429 0 R 431 0 R 433 0 R 435 0 R 437 0 R 439 0 R 441 0 R 443 0 R 445 0 R 447 0 R 449 0 R 451 0 R 453 0 R 455 0 R 457 0 R 459 0 R 461 0 R 463 0 R 465 0 R 467 0 R 469 0 R 471 0 R 473 0 R 475 0 R 477 0 R 479 0 R 481 0 R 483 0 R 485 0 R 487 0 R 489 0 R ] /Contents 382 0 R >> endobj 382 0 obj << /Length 33113 >> stream q 15.000 26.152 577.500 750.848 re W n 0.965 0.965 0.965 rg 26.250 687.232 555.000 89.768 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 687.232 m 581.250 687.232 l 581.250 687.982 l 26.250 687.982 l f q 35.250 698.482 537.000 78.518 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 8: BACHD behavioral measures derived from the computer vision as a function of genotype, age, and light )] TJ ET BT 35.250 755.571 Td /F4 9.8 Tf [(cycle)] TJ ET BT 35.250 736.201 Td /F1 9.8 Tf [(*: significant genotype effect for each age within each cycle; ***: significant genotype effect independent of cycle and age; #: )] TJ ET BT 35.250 722.465 Td /F1 9.8 Tf [(significant differences due to light cycle for each genotype and age; ##: significant differences due to light cycle for each )] TJ ET BT 35.250 708.729 Td /F1 9.8 Tf [(age independently of genotype; ###: significant differences due to light cycle independently of genotype and age.)] TJ ET Q BT 26.250 653.010 Td /F4 12.0 Tf [(Discussion)] TJ ET BT 26.250 633.056 Td /F1 9.8 Tf [(Here we demonstrated the utility of our novel high-throughput phenotyping system, PhenoCube)] TJ ET BT 437.037 636.944 Td /F1 8.7 Tf [(TM)] TJ ET BT 449.552 633.056 Td /F1 9.8 Tf [( by characterizing two )] TJ ET BT 26.250 621.151 Td /F1 9.8 Tf [(different genetic mouse models of HD. Our observations reveal the sensitivity and efficiency of this system in capturing )] TJ ET BT 26.250 609.246 Td /F1 9.8 Tf [(genotypic differences and tracking the course of the disease in multiple behavioral domains. Specifically, mice tested on this )] TJ ET BT 26.250 597.342 Td /F1 9.8 Tf [(platform exhibited robust differences across genotypes in cognitive performance, motor behavior, and circadian patterns )] TJ ET BT 26.250 585.437 Td /F1 9.8 Tf [(\(circadian data are described separately in )] TJ ET 0.267 0.267 0.267 rg BT 212.660 586.944 Td /F4 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 222.298 585.437 Td /F1 9.8 Tf [(\). These differences were captured by a fully automated system over a \(user-)] TJ ET BT 26.250 573.532 Td /F1 9.8 Tf [(specified\) wide time window and age range using electronic behavioral actuators \(chip reader and antenna from Intellicage\) as )] TJ ET BT 26.250 561.627 Td /F1 9.8 Tf [(well as computer vision tools.)] TJ ET BT 26.250 542.223 Td /F1 9.8 Tf [(Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 89.118 543.730 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 98.755 542.223 Td /F1 9.8 Tf [( assessed spatial alternation performance of R6/2 mice from the age of 8 weeks, primarily via patrolling task )] TJ ET BT 26.250 530.318 Td /F1 9.8 Tf [(where the position of the correct corner was changed in a clock-wise manner following a visit to a correct corner and at least )] TJ ET BT 26.250 518.413 Td /F1 9.8 Tf [(one nose-poke in that corner. They observed that R6/2 mice made significantly more place errors than did WT mice. Their )] TJ ET BT 26.250 506.508 Td /F1 9.8 Tf [(findings parallel the decreased proportion of alternation and increased proportion of repeat visits detected as early as 7 weeks )] TJ ET BT 26.250 494.604 Td /F1 9.8 Tf [(of age in our study. Importantly, perseverative responding has also been observed in HD patients as assessed in different tasks )] TJ ET 0.267 0.267 0.267 rg BT 26.250 484.206 Td /F4 8.7 Tf [(26)] TJ ET 0.271 0.267 0.267 rg BT 35.887 486.587 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 38.297 484.206 Td /F4 8.7 Tf [(27)] TJ ET 0.271 0.267 0.267 rg BT 47.934 486.587 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 50.343 484.206 Td /F4 8.7 Tf [(28)] TJ ET 0.271 0.267 0.267 rg BT 59.981 486.587 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 62.390 484.206 Td /F4 8.7 Tf [(29)] TJ ET 0.271 0.267 0.267 rg BT 72.027 482.699 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 463.294 Td /F1 9.8 Tf [(Relative to the R6/2 mouse, the BACHD model is not so well-characterized in terms of associated cognitive deficits, although )] TJ ET BT 26.250 451.389 Td /F1 9.8 Tf [(they have been shown to have normal response acquisition in an operant task and )] TJ ET 0.267 0.267 0.267 rg BT 384.485 452.897 Td /F4 8.7 Tf [(30)] TJ ET 0.271 0.267 0.267 rg BT 394.122 451.389 Td /F1 9.8 Tf [( and motivational deficits in a progressive )] TJ ET BT 26.250 439.485 Td /F1 9.8 Tf [(ratio task )] TJ ET 0.267 0.267 0.267 rg BT 68.516 440.992 Td /F4 8.7 Tf [(31)] TJ ET 0.271 0.267 0.267 rg BT 78.154 443.373 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 80.563 440.992 Td /F4 8.7 Tf [(20)] TJ ET 0.271 0.267 0.267 rg BT 90.200 439.485 Td /F1 9.8 Tf [( stated that, in a procedural task, BACHD mice showed a deficit during reversal but not acquisition. The )] TJ ET BT 26.250 427.580 Td /F1 9.8 Tf [(performance of the control mice in their study, however, was rather variable being close to chance levels at the end of both )] TJ ET BT 26.250 415.675 Td /F1 9.8 Tf [(training and testing. In our unpublished studies we have seen both deficits during acquisition and reversal in a 2 choice swim )] TJ ET BT 26.250 403.770 Td /F1 9.8 Tf [(task, but the results were neither consistent nor robust, suggesting that a possible deficit could be uncovered with a task that )] TJ ET BT 26.250 391.866 Td /F1 9.8 Tf [(elicits more stable performance in both the transgenic and control groups. While the BACHD mice in this study, like the R6/2 )] TJ ET BT 26.250 379.961 Td /F1 9.8 Tf [(mice, initially did appear to show stable tendency to make more repeat visits than controls, this phenotype declined with )] TJ ET BT 26.250 368.056 Td /F1 9.8 Tf [(increasing age, with significant increases in repeat visits seen only at 16 and 24 weeks, with a reduced rate of repeat visiting )] TJ ET BT 26.250 356.151 Td /F1 9.8 Tf [(detected at 52 weeks, while the R6/2 phenotype remained stable with increasing age.)] TJ ET BT 26.250 336.747 Td /F1 9.8 Tf [(The R6/2 mice also revealed an increase in drinking behavior relative to WT controls during the dark phase between 7-9 weeks )] TJ ET BT 26.250 324.842 Td /F1 9.8 Tf [(of age, whereas there were no significant differences during the light cycle. This result is also consistent with Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 558.941 326.349 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 568.579 324.842 Td /F1 9.8 Tf [(, )] TJ ET BT 26.250 312.937 Td /F1 9.8 Tf [(who observed significant differences in the frequency of licks between 7-9 weeks of age only during the active period )] TJ ET BT 26.250 301.032 Td /F1 9.8 Tf [(\(containing the entire dark phase and the first half of the light phase\). Different from their results however, the genotypic )] TJ ET BT 26.250 289.128 Td /F1 9.8 Tf [(difference in drinking behavior disappeared at 10 weeks of age in our study essentially due to a decrease in the lick frequency )] TJ ET BT 26.250 277.223 Td /F1 9.8 Tf [(of R6/2 mice, particularly during the dark phase. Note that IntelliCage does not measure the amount of liquid consumption but )] TJ ET BT 26.250 265.318 Td /F1 9.8 Tf [(rather the number and duration of licking behavior. Thus, we used the number of licks as a proxy for amount of water consumed )] TJ ET BT 26.250 253.413 Td /F1 9.8 Tf [(by mice \(see also )] TJ ET 0.267 0.267 0.267 rg BT 104.816 254.921 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 114.453 253.413 Td /F1 9.8 Tf [(\).)] TJ ET BT 26.250 234.009 Td /F1 9.8 Tf [(There were no differences between genotypes in terms of the total number of corner visits at least during their test ages \(i.e., 7-)] TJ ET BT 26.250 222.104 Td /F1 9.8 Tf [(10 weeks of age\). This result is also consistent with Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 313.456 223.611 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 323.093 222.104 Td /F1 9.8 Tf [( who observed significant decreases in corner visit )] TJ ET BT 26.250 210.199 Td /F1 9.8 Tf [(frequency starting from 11-12 weeks of age. Efficiency in collecting rewards \(operationalized as nose poking in correct corner )] TJ ET BT 26.250 198.294 Td /F1 9.8 Tf [(visits\) was equivalent in WT and R6/2 mice. The rate of such trials suggests that irrespective of genotype, our mouse subjects )] TJ ET BT 26.250 186.390 Td /F1 9.8 Tf [(had a weak tendency to explore when in the correct corners. Overall, we had the opportunity to compare our results with )] TJ ET BT 26.250 174.485 Td /F1 9.8 Tf [(previous findings \(i.e., )] TJ ET 0.267 0.267 0.267 rg BT 123.789 175.992 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 133.426 174.485 Td /F1 9.8 Tf [(\) in the case of R6/2 mice, which revealed a high degree of correspondence between studies.)] TJ ET BT 26.250 155.080 Td /F1 9.8 Tf [(BACHD animals exhibited significantly reduced numbers of corner visits but significant increases in their tendency to make )] TJ ET BT 26.250 143.175 Td /F1 9.8 Tf [(water seeking nosepoke responses when entering a correct corner compared to WT controls. Note that this increase in )] TJ ET BT 26.250 131.271 Td /F1 9.8 Tf [(efficiency of reward collection does not necessarily indicate enhanced cognitive performance of BACHD mice but rather the )] TJ ET BT 26.250 119.366 Td /F1 9.8 Tf [(suppression of alternative behaviors such as exploration and curiosity \(see also )] TJ ET 0.267 0.267 0.267 rg BT 371.439 120.873 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 381.076 119.366 Td /F1 9.8 Tf [(\), since the BACHD animals may only visit )] TJ ET BT 26.250 107.461 Td /F1 9.8 Tf [(corners in order to retrieve water. In addition, our study using the progressive ration task, suggests that incentive motivation )] TJ ET BT 26.250 95.556 Td /F1 9.8 Tf [(\(which energizes behavior in order to obtain reward\) is deficient in the BACHD mice )] TJ ET 0.267 0.267 0.267 rg BT 389.291 97.064 Td /F4 8.7 Tf [(31)] TJ ET 0.271 0.267 0.267 rg BT 398.929 95.556 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 76.152 Td /F1 9.8 Tf [(The BACHD mice exhibited increases in drinking behavior relative to WT controls, though this phenotype disappeared at the )] TJ ET BT 26.250 64.247 Td /F1 9.8 Tf [(final age of testing \(52 weeks of age\) due to a decrease in the lick frequency of BACHD mice, particularly during the dark phase. )] TJ ET BT 26.250 52.342 Td /F1 9.8 Tf [(These findings regarding drinking behavior are consistent with previously reported results from the R6/2 mice )] TJ ET 0.267 0.267 0.267 rg BT 498.764 53.849 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 508.402 56.230 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 510.811 53.849 Td /F4 8.7 Tf [(32)] TJ ET 0.271 0.267 0.267 rg BT 520.448 52.342 Td /F1 9.8 Tf [(. Increased )] TJ ET BT 26.250 40.437 Td /F1 9.8 Tf [(thirst has also been shown to characterize the HD progression in the clinic )] TJ ET 0.267 0.267 0.267 rg BT 348.673 41.945 Td /F4 8.7 Tf [(32)] TJ ET 0.271 0.267 0.267 rg BT 358.310 40.437 Td /F1 9.8 Tf [(. Importantly, perseverative responding has also )] TJ ET Q q 15.000 26.152 577.500 750.848 re W n 0.965 0.965 0.965 rg 26.250 687.232 555.000 89.768 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 687.232 m 581.250 687.232 l 581.250 687.982 l 26.250 687.982 l f q 35.250 698.482 537.000 78.518 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 8: BACHD behavioral measures derived from the computer vision as a function of genotype, age, and light )] TJ ET BT 35.250 755.571 Td /F4 9.8 Tf [(cycle)] TJ ET BT 35.250 736.201 Td /F1 9.8 Tf [(*: significant genotype effect for each age within each cycle; ***: significant genotype effect independent of cycle and age; #: )] TJ ET BT 35.250 722.465 Td /F1 9.8 Tf [(significant differences due to light cycle for each genotype and age; ##: significant differences due to light cycle for each )] TJ ET BT 35.250 708.729 Td /F1 9.8 Tf [(age independently of genotype; ###: significant differences due to light cycle independently of genotype and age.)] TJ ET Q BT 26.250 653.010 Td /F4 12.0 Tf [(Discussion)] TJ ET BT 26.250 633.056 Td /F1 9.8 Tf [(Here we demonstrated the utility of our novel high-throughput phenotyping system, PhenoCube)] TJ ET BT 437.037 636.944 Td /F1 8.7 Tf [(TM)] TJ ET BT 449.552 633.056 Td /F1 9.8 Tf [( by characterizing two )] TJ ET BT 26.250 621.151 Td /F1 9.8 Tf [(different genetic mouse models of HD. Our observations reveal the sensitivity and efficiency of this system in capturing )] TJ ET BT 26.250 609.246 Td /F1 9.8 Tf [(genotypic differences and tracking the course of the disease in multiple behavioral domains. Specifically, mice tested on this )] TJ ET BT 26.250 597.342 Td /F1 9.8 Tf [(platform exhibited robust differences across genotypes in cognitive performance, motor behavior, and circadian patterns )] TJ ET BT 26.250 585.437 Td /F1 9.8 Tf [(\(circadian data are described separately in )] TJ ET 0.267 0.267 0.267 rg BT 212.660 586.944 Td /F4 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 222.298 585.437 Td /F1 9.8 Tf [(\). These differences were captured by a fully automated system over a \(user-)] TJ ET BT 26.250 573.532 Td /F1 9.8 Tf [(specified\) wide time window and age range using electronic behavioral actuators \(chip reader and antenna from Intellicage\) as )] TJ ET BT 26.250 561.627 Td /F1 9.8 Tf [(well as computer vision tools.)] TJ ET BT 26.250 542.223 Td /F1 9.8 Tf [(Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 89.118 543.730 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 98.755 542.223 Td /F1 9.8 Tf [( assessed spatial alternation performance of R6/2 mice from the age of 8 weeks, primarily via patrolling task )] TJ ET BT 26.250 530.318 Td /F1 9.8 Tf [(where the position of the correct corner was changed in a clock-wise manner following a visit to a correct corner and at least )] TJ ET BT 26.250 518.413 Td /F1 9.8 Tf [(one nose-poke in that corner. They observed that R6/2 mice made significantly more place errors than did WT mice. Their )] TJ ET BT 26.250 506.508 Td /F1 9.8 Tf [(findings parallel the decreased proportion of alternation and increased proportion of repeat visits detected as early as 7 weeks )] TJ ET BT 26.250 494.604 Td /F1 9.8 Tf [(of age in our study. Importantly, perseverative responding has also been observed in HD patients as assessed in different tasks )] TJ ET 0.267 0.267 0.267 rg BT 26.250 484.206 Td /F4 8.7 Tf [(26)] TJ ET 0.271 0.267 0.267 rg BT 35.887 486.587 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 38.297 484.206 Td /F4 8.7 Tf [(27)] TJ ET 0.271 0.267 0.267 rg BT 47.934 486.587 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 50.343 484.206 Td /F4 8.7 Tf [(28)] TJ ET 0.271 0.267 0.267 rg BT 59.981 486.587 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 62.390 484.206 Td /F4 8.7 Tf [(29)] TJ ET 0.271 0.267 0.267 rg BT 72.027 482.699 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 463.294 Td /F1 9.8 Tf [(Relative to the R6/2 mouse, the BACHD model is not so well-characterized in terms of associated cognitive deficits, although )] TJ ET BT 26.250 451.389 Td /F1 9.8 Tf [(they have been shown to have normal response acquisition in an operant task and )] TJ ET 0.267 0.267 0.267 rg BT 384.485 452.897 Td /F4 8.7 Tf [(30)] TJ ET 0.271 0.267 0.267 rg BT 394.122 451.389 Td /F1 9.8 Tf [( and motivational deficits in a progressive )] TJ ET BT 26.250 439.485 Td /F1 9.8 Tf [(ratio task )] TJ ET 0.267 0.267 0.267 rg BT 68.516 440.992 Td /F4 8.7 Tf [(31)] TJ ET 0.271 0.267 0.267 rg BT 78.154 443.373 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 80.563 440.992 Td /F4 8.7 Tf [(20)] TJ ET 0.271 0.267 0.267 rg BT 90.200 439.485 Td /F1 9.8 Tf [( stated that, in a procedural task, BACHD mice showed a deficit during reversal but not acquisition. The )] TJ ET BT 26.250 427.580 Td /F1 9.8 Tf [(performance of the control mice in their study, however, was rather variable being close to chance levels at the end of both )] TJ ET BT 26.250 415.675 Td /F1 9.8 Tf [(training and testing. In our unpublished studies we have seen both deficits during acquisition and reversal in a 2 choice swim )] TJ ET BT 26.250 403.770 Td /F1 9.8 Tf [(task, but the results were neither consistent nor robust, suggesting that a possible deficit could be uncovered with a task that )] TJ ET BT 26.250 391.866 Td /F1 9.8 Tf [(elicits more stable performance in both the transgenic and control groups. While the BACHD mice in this study, like the R6/2 )] TJ ET BT 26.250 379.961 Td /F1 9.8 Tf [(mice, initially did appear to show stable tendency to make more repeat visits than controls, this phenotype declined with )] TJ ET BT 26.250 368.056 Td /F1 9.8 Tf [(increasing age, with significant increases in repeat visits seen only at 16 and 24 weeks, with a reduced rate of repeat visiting )] TJ ET BT 26.250 356.151 Td /F1 9.8 Tf [(detected at 52 weeks, while the R6/2 phenotype remained stable with increasing age.)] TJ ET BT 26.250 336.747 Td /F1 9.8 Tf [(The R6/2 mice also revealed an increase in drinking behavior relative to WT controls during the dark phase between 7-9 weeks )] TJ ET BT 26.250 324.842 Td /F1 9.8 Tf [(of age, whereas there were no significant differences during the light cycle. This result is also consistent with Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 558.941 326.349 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 568.579 324.842 Td /F1 9.8 Tf [(, )] TJ ET BT 26.250 312.937 Td /F1 9.8 Tf [(who observed significant differences in the frequency of licks between 7-9 weeks of age only during the active period )] TJ ET BT 26.250 301.032 Td /F1 9.8 Tf [(\(containing the entire dark phase and the first half of the light phase\). Different from their results however, the genotypic )] TJ ET BT 26.250 289.128 Td /F1 9.8 Tf [(difference in drinking behavior disappeared at 10 weeks of age in our study essentially due to a decrease in the lick frequency )] TJ ET BT 26.250 277.223 Td /F1 9.8 Tf [(of R6/2 mice, particularly during the dark phase. Note that IntelliCage does not measure the amount of liquid consumption but )] TJ ET BT 26.250 265.318 Td /F1 9.8 Tf [(rather the number and duration of licking behavior. Thus, we used the number of licks as a proxy for amount of water consumed )] TJ ET BT 26.250 253.413 Td /F1 9.8 Tf [(by mice \(see also )] TJ ET 0.267 0.267 0.267 rg BT 104.816 254.921 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 114.453 253.413 Td /F1 9.8 Tf [(\).)] TJ ET BT 26.250 234.009 Td /F1 9.8 Tf [(There were no differences between genotypes in terms of the total number of corner visits at least during their test ages \(i.e., 7-)] TJ ET BT 26.250 222.104 Td /F1 9.8 Tf [(10 weeks of age\). This result is also consistent with Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 313.456 223.611 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 323.093 222.104 Td /F1 9.8 Tf [( who observed significant decreases in corner visit )] TJ ET BT 26.250 210.199 Td /F1 9.8 Tf [(frequency starting from 11-12 weeks of age. Efficiency in collecting rewards \(operationalized as nose poking in correct corner )] TJ ET BT 26.250 198.294 Td /F1 9.8 Tf [(visits\) was equivalent in WT and R6/2 mice. The rate of such trials suggests that irrespective of genotype, our mouse subjects )] TJ ET BT 26.250 186.390 Td /F1 9.8 Tf [(had a weak tendency to explore when in the correct corners. Overall, we had the opportunity to compare our results with )] TJ ET BT 26.250 174.485 Td /F1 9.8 Tf [(previous findings \(i.e., )] TJ ET 0.267 0.267 0.267 rg BT 123.789 175.992 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 133.426 174.485 Td /F1 9.8 Tf [(\) in the case of R6/2 mice, which revealed a high degree of correspondence between studies.)] TJ ET BT 26.250 155.080 Td /F1 9.8 Tf [(BACHD animals exhibited significantly reduced numbers of corner visits but significant increases in their tendency to make )] TJ ET BT 26.250 143.175 Td /F1 9.8 Tf [(water seeking nosepoke responses when entering a correct corner compared to WT controls. Note that this increase in )] TJ ET BT 26.250 131.271 Td /F1 9.8 Tf [(efficiency of reward collection does not necessarily indicate enhanced cognitive performance of BACHD mice but rather the )] TJ ET BT 26.250 119.366 Td /F1 9.8 Tf [(suppression of alternative behaviors such as exploration and curiosity \(see also )] TJ ET 0.267 0.267 0.267 rg BT 371.439 120.873 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 381.076 119.366 Td /F1 9.8 Tf [(\), since the BACHD animals may only visit )] TJ ET BT 26.250 107.461 Td /F1 9.8 Tf [(corners in order to retrieve water. In addition, our study using the progressive ration task, suggests that incentive motivation )] TJ ET BT 26.250 95.556 Td /F1 9.8 Tf [(\(which energizes behavior in order to obtain reward\) is deficient in the BACHD mice )] TJ ET 0.267 0.267 0.267 rg BT 389.291 97.064 Td /F4 8.7 Tf [(31)] TJ ET 0.271 0.267 0.267 rg BT 398.929 95.556 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 76.152 Td /F1 9.8 Tf [(The BACHD mice exhibited increases in drinking behavior relative to WT controls, though this phenotype disappeared at the )] TJ ET BT 26.250 64.247 Td /F1 9.8 Tf [(final age of testing \(52 weeks of age\) due to a decrease in the lick frequency of BACHD mice, particularly during the dark phase. )] TJ ET BT 26.250 52.342 Td /F1 9.8 Tf [(These findings regarding drinking behavior are consistent with previously reported results from the R6/2 mice )] TJ ET 0.267 0.267 0.267 rg BT 498.764 53.849 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 508.402 56.230 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 510.811 53.849 Td /F4 8.7 Tf [(32)] TJ ET 0.271 0.267 0.267 rg BT 520.448 52.342 Td /F1 9.8 Tf [(. Increased )] TJ ET BT 26.250 40.437 Td /F1 9.8 Tf [(thirst has also been shown to characterize the HD progression in the clinic )] TJ ET 0.267 0.267 0.267 rg BT 348.673 41.945 Td /F4 8.7 Tf [(32)] TJ ET 0.271 0.267 0.267 rg BT 358.310 40.437 Td /F1 9.8 Tf [(. Importantly, perseverative responding has also )] TJ ET Q q 15.000 26.152 577.500 750.848 re W n 0.965 0.965 0.965 rg 26.250 687.232 555.000 89.768 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 687.232 m 581.250 687.232 l 581.250 687.982 l 26.250 687.982 l f q 35.250 698.482 537.000 78.518 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F4 9.8 Tf [(Fig. 8: BACHD behavioral measures derived from the computer vision as a function of genotype, age, and light )] TJ ET BT 35.250 755.571 Td /F4 9.8 Tf [(cycle)] TJ ET BT 35.250 736.201 Td /F1 9.8 Tf [(*: significant genotype effect for each age within each cycle; ***: significant genotype effect independent of cycle and age; #: )] TJ ET BT 35.250 722.465 Td /F1 9.8 Tf [(significant differences due to light cycle for each genotype and age; ##: significant differences due to light cycle for each )] TJ ET BT 35.250 708.729 Td /F1 9.8 Tf [(age independently of genotype; ###: significant differences due to light cycle independently of genotype and age.)] TJ ET Q BT 26.250 653.010 Td /F4 12.0 Tf [(Discussion)] TJ ET BT 26.250 633.056 Td /F1 9.8 Tf [(Here we demonstrated the utility of our novel high-throughput phenotyping system, PhenoCube)] TJ ET BT 437.037 636.944 Td /F1 8.7 Tf [(TM)] TJ ET BT 449.552 633.056 Td /F1 9.8 Tf [( by characterizing two )] TJ ET BT 26.250 621.151 Td /F1 9.8 Tf [(different genetic mouse models of HD. Our observations reveal the sensitivity and efficiency of this system in capturing )] TJ ET BT 26.250 609.246 Td /F1 9.8 Tf [(genotypic differences and tracking the course of the disease in multiple behavioral domains. Specifically, mice tested on this )] TJ ET BT 26.250 597.342 Td /F1 9.8 Tf [(platform exhibited robust differences across genotypes in cognitive performance, motor behavior, and circadian patterns )] TJ ET BT 26.250 585.437 Td /F1 9.8 Tf [(\(circadian data are described separately in )] TJ ET 0.267 0.267 0.267 rg BT 212.660 586.944 Td /F4 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 222.298 585.437 Td /F1 9.8 Tf [(\). These differences were captured by a fully automated system over a \(user-)] TJ ET BT 26.250 573.532 Td /F1 9.8 Tf [(specified\) wide time window and age range using electronic behavioral actuators \(chip reader and antenna from Intellicage\) as )] TJ ET BT 26.250 561.627 Td /F1 9.8 Tf [(well as computer vision tools.)] TJ ET BT 26.250 542.223 Td /F1 9.8 Tf [(Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 89.118 543.730 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 98.755 542.223 Td /F1 9.8 Tf [( assessed spatial alternation performance of R6/2 mice from the age of 8 weeks, primarily via patrolling task )] TJ ET BT 26.250 530.318 Td /F1 9.8 Tf [(where the position of the correct corner was changed in a clock-wise manner following a visit to a correct corner and at least )] TJ ET BT 26.250 518.413 Td /F1 9.8 Tf [(one nose-poke in that corner. They observed that R6/2 mice made significantly more place errors than did WT mice. Their )] TJ ET BT 26.250 506.508 Td /F1 9.8 Tf [(findings parallel the decreased proportion of alternation and increased proportion of repeat visits detected as early as 7 weeks )] TJ ET BT 26.250 494.604 Td /F1 9.8 Tf [(of age in our study. Importantly, perseverative responding has also been observed in HD patients as assessed in different tasks )] TJ ET 0.267 0.267 0.267 rg BT 26.250 484.206 Td /F4 8.7 Tf [(26)] TJ ET 0.271 0.267 0.267 rg BT 35.887 486.587 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 38.297 484.206 Td /F4 8.7 Tf [(27)] TJ ET 0.271 0.267 0.267 rg BT 47.934 486.587 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 50.343 484.206 Td /F4 8.7 Tf [(28)] TJ ET 0.271 0.267 0.267 rg BT 59.981 486.587 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 62.390 484.206 Td /F4 8.7 Tf [(29)] TJ ET 0.271 0.267 0.267 rg BT 72.027 482.699 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 463.294 Td /F1 9.8 Tf [(Relative to the R6/2 mouse, the BACHD model is not so well-characterized in terms of associated cognitive deficits, although )] TJ ET BT 26.250 451.389 Td /F1 9.8 Tf [(they have been shown to have normal response acquisition in an operant task and )] TJ ET 0.267 0.267 0.267 rg BT 384.485 452.897 Td /F4 8.7 Tf [(30)] TJ ET 0.271 0.267 0.267 rg BT 394.122 451.389 Td /F1 9.8 Tf [( and motivational deficits in a progressive )] TJ ET BT 26.250 439.485 Td /F1 9.8 Tf [(ratio task )] TJ ET 0.267 0.267 0.267 rg BT 68.516 440.992 Td /F4 8.7 Tf [(31)] TJ ET 0.271 0.267 0.267 rg BT 78.154 443.373 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 80.563 440.992 Td /F4 8.7 Tf [(20)] TJ ET 0.271 0.267 0.267 rg BT 90.200 439.485 Td /F1 9.8 Tf [( stated that, in a procedural task, BACHD mice showed a deficit during reversal but not acquisition. The )] TJ ET BT 26.250 427.580 Td /F1 9.8 Tf [(performance of the control mice in their study, however, was rather variable being close to chance levels at the end of both )] TJ ET BT 26.250 415.675 Td /F1 9.8 Tf [(training and testing. In our unpublished studies we have seen both deficits during acquisition and reversal in a 2 choice swim )] TJ ET BT 26.250 403.770 Td /F1 9.8 Tf [(task, but the results were neither consistent nor robust, suggesting that a possible deficit could be uncovered with a task that )] TJ ET BT 26.250 391.866 Td /F1 9.8 Tf [(elicits more stable performance in both the transgenic and control groups. While the BACHD mice in this study, like the R6/2 )] TJ ET BT 26.250 379.961 Td /F1 9.8 Tf [(mice, initially did appear to show stable tendency to make more repeat visits than controls, this phenotype declined with )] TJ ET BT 26.250 368.056 Td /F1 9.8 Tf [(increasing age, with significant increases in repeat visits seen only at 16 and 24 weeks, with a reduced rate of repeat visiting )] TJ ET BT 26.250 356.151 Td /F1 9.8 Tf [(detected at 52 weeks, while the R6/2 phenotype remained stable with increasing age.)] TJ ET BT 26.250 336.747 Td /F1 9.8 Tf [(The R6/2 mice also revealed an increase in drinking behavior relative to WT controls during the dark phase between 7-9 weeks )] TJ ET BT 26.250 324.842 Td /F1 9.8 Tf [(of age, whereas there were no significant differences during the light cycle. This result is also consistent with Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 558.941 326.349 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 568.579 324.842 Td /F1 9.8 Tf [(, )] TJ ET BT 26.250 312.937 Td /F1 9.8 Tf [(who observed significant differences in the frequency of licks between 7-9 weeks of age only during the active period )] TJ ET BT 26.250 301.032 Td /F1 9.8 Tf [(\(containing the entire dark phase and the first half of the light phase\). Different from their results however, the genotypic )] TJ ET BT 26.250 289.128 Td /F1 9.8 Tf [(difference in drinking behavior disappeared at 10 weeks of age in our study essentially due to a decrease in the lick frequency )] TJ ET BT 26.250 277.223 Td /F1 9.8 Tf [(of R6/2 mice, particularly during the dark phase. Note that IntelliCage does not measure the amount of liquid consumption but )] TJ ET BT 26.250 265.318 Td /F1 9.8 Tf [(rather the number and duration of licking behavior. Thus, we used the number of licks as a proxy for amount of water consumed )] TJ ET BT 26.250 253.413 Td /F1 9.8 Tf [(by mice \(see also )] TJ ET 0.267 0.267 0.267 rg BT 104.816 254.921 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 114.453 253.413 Td /F1 9.8 Tf [(\).)] TJ ET BT 26.250 234.009 Td /F1 9.8 Tf [(There were no differences between genotypes in terms of the total number of corner visits at least during their test ages \(i.e., 7-)] TJ ET BT 26.250 222.104 Td /F1 9.8 Tf [(10 weeks of age\). This result is also consistent with Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 313.456 223.611 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 323.093 222.104 Td /F1 9.8 Tf [( who observed significant decreases in corner visit )] TJ ET BT 26.250 210.199 Td /F1 9.8 Tf [(frequency starting from 11-12 weeks of age. Efficiency in collecting rewards \(operationalized as nose poking in correct corner )] TJ ET BT 26.250 198.294 Td /F1 9.8 Tf [(visits\) was equivalent in WT and R6/2 mice. The rate of such trials suggests that irrespective of genotype, our mouse subjects )] TJ ET BT 26.250 186.390 Td /F1 9.8 Tf [(had a weak tendency to explore when in the correct corners. Overall, we had the opportunity to compare our results with )] TJ ET BT 26.250 174.485 Td /F1 9.8 Tf [(previous findings \(i.e., )] TJ ET 0.267 0.267 0.267 rg BT 123.789 175.992 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 133.426 174.485 Td /F1 9.8 Tf [(\) in the case of R6/2 mice, which revealed a high degree of correspondence between studies.)] TJ ET BT 26.250 155.080 Td /F1 9.8 Tf [(BACHD animals exhibited significantly reduced numbers of corner visits but significant increases in their tendency to make )] TJ ET BT 26.250 143.175 Td /F1 9.8 Tf [(water seeking nosepoke responses when entering a correct corner compared to WT controls. Note that this increase in )] TJ ET BT 26.250 131.271 Td /F1 9.8 Tf [(efficiency of reward collection does not necessarily indicate enhanced cognitive performance of BACHD mice but rather the )] TJ ET BT 26.250 119.366 Td /F1 9.8 Tf [(suppression of alternative behaviors such as exploration and curiosity \(see also )] TJ ET 0.267 0.267 0.267 rg BT 371.439 120.873 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 381.076 119.366 Td /F1 9.8 Tf [(\), since the BACHD animals may only visit )] TJ ET BT 26.250 107.461 Td /F1 9.8 Tf [(corners in order to retrieve water. In addition, our study using the progressive ration task, suggests that incentive motivation )] TJ ET BT 26.250 95.556 Td /F1 9.8 Tf [(\(which energizes behavior in order to obtain reward\) is deficient in the BACHD mice )] TJ ET 0.267 0.267 0.267 rg BT 389.291 97.064 Td /F4 8.7 Tf [(31)] TJ ET 0.271 0.267 0.267 rg BT 398.929 95.556 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 76.152 Td /F1 9.8 Tf [(The BACHD mice exhibited increases in drinking behavior relative to WT controls, though this phenotype disappeared at the )] TJ ET BT 26.250 64.247 Td /F1 9.8 Tf [(final age of testing \(52 weeks of age\) due to a decrease in the lick frequency of BACHD mice, particularly during the dark phase. )] TJ ET BT 26.250 52.342 Td /F1 9.8 Tf [(These findings regarding drinking behavior are consistent with previously reported results from the R6/2 mice )] TJ ET 0.267 0.267 0.267 rg BT 498.764 53.849 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 508.402 56.230 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 510.811 53.849 Td /F4 8.7 Tf [(32)] TJ ET 0.271 0.267 0.267 rg BT 520.448 52.342 Td /F1 9.8 Tf [(. Increased )] TJ ET BT 26.250 40.437 Td /F1 9.8 Tf [(thirst has also been shown to characterize the HD progression in the clinic )] TJ ET 0.267 0.267 0.267 rg BT 348.673 41.945 Td /F4 8.7 Tf [(32)] TJ ET 0.271 0.267 0.267 rg BT 358.310 40.437 Td /F1 9.8 Tf [(. Importantly, perseverative responding has also )] TJ ET Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(10)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Huntington Disease)] TJ ET Q endstream endobj 383 0 obj << /Type /Annot /Subtype /Link /A 384 0 R /Border [0 0 0] /H /I /Rect [ 212.6602 586.1424 222.2976 594.9607 ] >> endobj 384 0 obj << /Type /Action >> endobj 385 0 obj << /Type /Annot /Subtype /Link /A 386 0 R /Border [0 0 0] /H /I /Rect [ 89.1180 542.9282 98.7553 551.7465 ] >> endobj 386 0 obj << /Type /Action >> endobj 387 0 obj << /Type /Annot /Subtype /Link /A 388 0 R /Border [0 0 0] /H /I /Rect [ 26.2500 483.4044 35.8873 492.2227 ] >> endobj 388 0 obj << /Type /Action >> endobj 389 0 obj << /Type /Annot /Subtype /Link /A 390 0 R /Border [0 0 0] /H /I /Rect [ 38.2967 483.4044 47.9340 492.2227 ] >> endobj 390 0 obj << /Type /Action >> endobj 391 0 obj << /Type /Annot /Subtype /Link /A 392 0 R /Border [0 0 0] /H /I /Rect [ 50.3433 483.4044 59.9807 492.2227 ] 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rg BT 300.045 768.983 Td /F4 8.7 Tf [(27)] TJ ET 0.271 0.267 0.267 rg BT 309.683 771.364 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 312.092 768.983 Td /F4 8.7 Tf [(28)] TJ ET 0.271 0.267 0.267 rg BT 321.729 771.364 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 324.139 768.983 Td /F4 8.7 Tf [(29)] TJ ET 0.271 0.267 0.267 rg BT 333.776 767.476 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 748.071 Td /F1 9.8 Tf [(We also evaluated the motor behavior of the R6/2 and BACHD mice using the computer vision software revealing that maybe )] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(R6/2 but clearly BACHD mice had shorter total path length, \(not significant for R6/2 mice\), reduced levels of rearing and )] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(climbing behavior, and higher immobility time. These results are consistent with previous reports of decreased locomotion )] TJ ET 0.267 0.267 0.267 rg BT 552.399 725.769 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 557.218 728.150 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 559.627 725.769 Td /F4 8.7 Tf [(33)] TJ ET 0.271 0.267 0.267 rg BT 569.264 724.262 Td /F1 9.8 Tf [(, )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(rearing-climbing )] TJ ET 0.267 0.267 0.267 rg BT 98.302 713.864 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 103.121 712.357 Td /F1 9.8 Tf [(, and imbalance and motor coordination problems )] TJ ET 0.267 0.267 0.267 rg BT 319.883 713.864 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 324.702 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 327.111 713.864 Td /F4 8.7 Tf [(20)] TJ ET 0.271 0.267 0.267 rg BT 336.749 712.357 Td /F1 9.8 Tf [( in R6/2 and BACHD mice. The performance of R6/2 )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(mice particularly paralleled the results of Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 265.759 701.960 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 275.396 700.452 Td /F1 9.8 Tf [( who tested these mice on open field )] TJ ET 0.267 0.267 0.267 rg BT 436.359 701.960 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 441.178 704.341 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 443.587 701.960 Td /F4 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 453.224 700.452 Td /F1 9.8 Tf [(. They observed significant )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(decrease in number of rearings as early as 6 weeks of age, however differences in distance traveled occurred reliably at 12 )] TJ ET BT 26.250 676.643 Td /F1 9.8 Tf [(weeks of age. Light cycle was found to affect many of our computer vision measures, which has also been reported in studies )] TJ ET BT 26.250 664.738 Td /F1 9.8 Tf [(that tested WT mice in open field \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 195.871 666.245 Td /F4 8.7 Tf [(34)] TJ ET 0.271 0.267 0.267 rg BT 205.508 668.626 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 207.917 666.245 Td /F4 8.7 Tf [(35)] TJ ET 0.271 0.267 0.267 rg BT 217.555 664.738 Td /F1 9.8 Tf [( at least for C57BL/6\).)] TJ ET BT 26.250 645.333 Td /F1 9.8 Tf [(Although the differences in locomotion were in the expected direction, these differences were not statistically significant for the )] TJ ET BT 26.250 633.429 Td /F1 9.8 Tf [(R6/2 mice \()] TJ ET BT 76.092 633.429 Td /F5 9.8 Tf [(p)] TJ ET BT 81.513 633.429 Td /F1 9.8 Tf [(=.31\). On the other hand, same R6/2 mice were significantly more immobile than their wild-type controls. The )] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(dissociation between these two measures is possible since immobility and locomotion \(i.e., path length\) are not perfectly )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(complementary given all other alternative behaviors animals engage in \(e.g., grooming\). A more detailed characterization of )] TJ ET BT 26.250 597.714 Td /F1 9.8 Tf [(behavior via computer vision system in future studies can clarify the dynamics that might underlie this dissociation.)] TJ ET BT 26.250 578.310 Td /F1 9.8 Tf [(Since our computer vision application does not track the individual animal identities in a consistent manner \(see below for )] TJ ET BT 26.250 566.405 Td /F1 9.8 Tf [(details\), in order to compare genotypes in terms of computer vision indices, mice with the same genotype were housed in the )] TJ ET BT 26.250 554.500 Td /F1 9.8 Tf [(same cage. While, we could track the identity of a given mouse prior to and following corner exits \(based on time-stamped )] TJ ET BT 26.250 542.595 Td /F1 9.8 Tf [(transponder detections\), these periods of identity tracking were constrained by instances of occlusions, where identity swaps )] TJ ET BT 26.250 530.691 Td /F1 9.8 Tf [(are likely. Ongoing development of the computer vision application is focused on reliable individual tracking and social behavior. )] TJ ET BT 26.250 518.786 Td /F1 9.8 Tf [(Another advantage of genotype-based housing in our specific case was to avoid possible dominance of the WT mice over R6/2 )] TJ ET BT 26.250 506.881 Td /F1 9.8 Tf [(and BACHD mice minimizing their access to resources. This would have constituted an important confound during behavioral )] TJ ET BT 26.250 494.976 Td /F1 9.8 Tf [(phenotyping of these mice. Certain behavioral phenotypes of R6/2 and BACHD mice \(e.g., disrupted sleep patterns\) could also )] TJ ET BT 26.250 483.072 Td /F1 9.8 Tf [(directly interact with WT mouse behavior. Similar factors however could also constitute a disadvantage of genotype-based )] TJ ET BT 26.250 471.167 Td /F1 9.8 Tf [(housing schemes as they could inflate the degree of behavioral phenotype. For instance, disrupted sleep pattern of a given )] TJ ET BT 26.250 459.262 Td /F1 9.8 Tf [(mouse could also disrupt the sleep patterns of other mice in the same cage.)] TJ ET BT 26.250 439.857 Td /F1 9.8 Tf [(Group housing mice during testing might itself exert some disadvantages. One of the likely problems with this approach is that )] TJ ET BT 26.250 427.953 Td /F1 9.8 Tf [(group housing might induce social ranking dynamics, which might in turn influence behavior and task-related performances. In )] TJ ET BT 26.250 416.048 Td /F1 9.8 Tf [(this study, we did not specifically evaluate social ranking and thus could not capture these possible relations. On the other hand, )] TJ ET BT 26.250 404.143 Td /F1 9.8 Tf [(in combination with marking animals for individuation, analytical tools can be developed to evaluate social behavioral dynamics )] TJ ET BT 26.250 392.238 Td /F1 9.8 Tf [(based on computer vision application endpoints. For instance, huddling behavior and/or dispersal of animals in the home-cage )] TJ ET BT 26.250 380.334 Td /F1 9.8 Tf [(might provide important data regarding social behavior. Genotype-based housing or group-housing itself exerts a trade-off that )] TJ ET BT 26.250 368.429 Td /F1 9.8 Tf [(needs to be evaluated given what is known about the mouse lines to be tested.)] TJ ET BT 26.250 349.024 Td /F1 9.8 Tf [(Another important issue is the number of subjects to test in a given case. In our case, the number of subjects was chosen to )] TJ ET BT 26.250 337.119 Td /F1 9.8 Tf [(maximize the sample size \(as allowed by regulations\). Power analysis shows that group sizes of 3-11 mice for most of the )] TJ ET BT 26.250 325.215 Td /F1 9.8 Tf [(parameters measured in the R6/2 mice at 8 weeks of age \(namely: corner entries, licking count, percentage of repeats, )] TJ ET BT 26.250 313.310 Td /F1 9.8 Tf [(percentage of alternations, percent time immobile and percent time rearing-climbing\) are sufficient to detect a 50% improvement )] TJ ET BT 26.250 301.405 Td /F1 9.8 Tf [(of a potential therapy \(final number depends on the parameter measured; ?= 0.05, 80% power\). A similar sample size allows )] TJ ET BT 26.250 289.500 Td /F1 9.8 Tf [(the evaluation of potential therapies at 10 weeks of age R6/2 mice in the PhenoCube)] TJ ET BT 392.080 293.389 Td /F1 8.7 Tf [(TM)] TJ ET BT 404.594 289.500 Td /F1 9.8 Tf [( system when measuring percent )] TJ ET BT 26.250 277.596 Td /F1 9.8 Tf [(alternations, percent time immobile and percent time rearing-climbing. In the BACHD mice, power analysis indicates that group )] TJ ET BT 26.250 265.691 Td /F1 9.8 Tf [(sizes of 4-16 mice are sufficient to detect a 50% effect improvement at 36 weeks of age when measuring corner entries, )] TJ ET BT 26.250 253.786 Td /F1 9.8 Tf [(collected rewards, and locomotion. If the effect of the potential therapy is examined at 52 weeks of age, group sizes of 4-9 mice )] TJ ET BT 26.250 241.881 Td /F1 9.8 Tf [(are needed when measuring corner entries, collected rewards, locomotion, percent time immobile and percent time rearing-)] TJ ET BT 26.250 229.977 Td /F1 9.8 Tf [(climbing.)] TJ ET BT 26.250 210.572 Td /F1 9.8 Tf [(One of the important features of the data gathered from this study is the increased licking that could be interpreted as increased )] TJ ET BT 26.250 198.667 Td /F1 9.8 Tf [(thirst of both R6/2 and BACHD mice coupled with the use of water as reward. Increased licking phenotype might constitute a )] TJ ET BT 26.250 186.762 Td /F1 9.8 Tf [(confound for behavioral tasks that set the rules for access to water reward )] TJ ET 0.267 0.267 0.267 rg BT 348.146 188.270 Td /F4 8.7 Tf [(32)] TJ ET 0.271 0.267 0.267 rg BT 357.784 186.762 Td /F1 9.8 Tf [(. That have been said, the canonical interpretation )] TJ ET BT 26.250 174.858 Td /F1 9.8 Tf [(of higher thirst would be increased motivation for water reward, which if anything would be expected to lead to more well-)] TJ ET BT 26.250 162.953 Td /F1 9.8 Tf [(structured \(and possibly less disrupted\) goal-directed behaviors. On the other hand, several phenotypes such as the frequency )] TJ ET BT 26.250 151.048 Td /F1 9.8 Tf [(of corner visits might have been masked by higher motivation for water reward at least in the R6/2 mice.)] TJ ET BT 26.250 131.643 Td /F1 9.8 Tf [(Note that the aim of this study was not only to characterize the disease profiles of two different mutations, but instead to )] TJ ET BT 26.250 119.739 Td /F1 9.8 Tf [(demonstrate the utility of the PhenoCube)] TJ ET BT 202.930 123.627 Td /F1 8.7 Tf [(TM)] TJ ET BT 215.444 119.739 Td /F1 9.8 Tf [( in mouse models of HD. Further work can test different mouse models at the same )] TJ ET BT 26.250 107.834 Td /F1 9.8 Tf [(age \(e.g., BACHD vs. R6/2 mice at 8 weeks of age\) with identical experimental protocols to enable a direct comparison of )] TJ ET BT 26.250 95.929 Td /F1 9.8 Tf [(mouse models of HD using PhenoCube)] TJ ET BT 197.489 99.817 Td /F1 8.7 Tf [(TM)] TJ ET BT 210.004 95.929 Td /F1 9.8 Tf [( as a tool. Another interesting possibility for future studies would be mixing different )] TJ ET BT 26.250 84.024 Td /F1 9.8 Tf [(genotypes in a cage and comparing their performance to conditions where mice of the same genotype are housed together \(as )] TJ ET BT 26.250 72.120 Td /F1 9.8 Tf [(in this study\). This would enable the differentiation of disease progression in the presence and absence of mice with HD and )] TJ ET BT 26.250 60.215 Td /F1 9.8 Tf [(might reveal possibly differential social behavior dynamics compared to genotype-based housing scheme. Finally, defining the )] TJ ET BT 26.250 48.310 Td /F1 9.8 Tf [(accuracy of nose-poke based on randomly signaled water recesses would constitute a test of cued discrimination performance, )] TJ ET BT 26.250 36.405 Td /F1 9.8 Tf [(which can be compared to the spatial discrimination of recesses as in the case of this study. These two different strategies are )] TJ ET Q q 15.000 22.120 577.500 754.880 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(been observed in HD patients as assessed in different tasks )] TJ ET 0.267 0.267 0.267 rg BT 287.998 768.983 Td /F4 8.7 Tf [(26)] TJ ET 0.271 0.267 0.267 rg BT 297.636 771.364 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 300.045 768.983 Td /F4 8.7 Tf [(27)] TJ ET 0.271 0.267 0.267 rg BT 309.683 771.364 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 312.092 768.983 Td /F4 8.7 Tf [(28)] TJ ET 0.271 0.267 0.267 rg BT 321.729 771.364 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 324.139 768.983 Td /F4 8.7 Tf [(29)] TJ ET 0.271 0.267 0.267 rg BT 333.776 767.476 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 748.071 Td /F1 9.8 Tf [(We also evaluated the motor behavior of the R6/2 and BACHD mice using the computer vision software revealing that maybe )] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(R6/2 but clearly BACHD mice had shorter total path length, \(not significant for R6/2 mice\), reduced levels of rearing and )] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(climbing behavior, and higher immobility time. These results are consistent with previous reports of decreased locomotion )] TJ ET 0.267 0.267 0.267 rg BT 552.399 725.769 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 557.218 728.150 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 559.627 725.769 Td /F4 8.7 Tf [(33)] TJ ET 0.271 0.267 0.267 rg BT 569.264 724.262 Td /F1 9.8 Tf [(, )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(rearing-climbing )] TJ ET 0.267 0.267 0.267 rg BT 98.302 713.864 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 103.121 712.357 Td /F1 9.8 Tf [(, and imbalance and motor coordination problems )] TJ ET 0.267 0.267 0.267 rg BT 319.883 713.864 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 324.702 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 327.111 713.864 Td /F4 8.7 Tf [(20)] TJ ET 0.271 0.267 0.267 rg BT 336.749 712.357 Td /F1 9.8 Tf [( in R6/2 and BACHD mice. The performance of R6/2 )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(mice particularly paralleled the results of Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 265.759 701.960 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 275.396 700.452 Td /F1 9.8 Tf [( who tested these mice on open field )] TJ ET 0.267 0.267 0.267 rg BT 436.359 701.960 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 441.178 704.341 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 443.587 701.960 Td /F4 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 453.224 700.452 Td /F1 9.8 Tf [(. They observed significant )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(decrease in number of rearings as early as 6 weeks of age, however differences in distance traveled occurred reliably at 12 )] TJ ET BT 26.250 676.643 Td /F1 9.8 Tf [(weeks of age. Light cycle was found to affect many of our computer vision measures, which has also been reported in studies )] TJ ET BT 26.250 664.738 Td /F1 9.8 Tf [(that tested WT mice in open field \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 195.871 666.245 Td /F4 8.7 Tf [(34)] TJ ET 0.271 0.267 0.267 rg BT 205.508 668.626 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 207.917 666.245 Td /F4 8.7 Tf [(35)] TJ ET 0.271 0.267 0.267 rg BT 217.555 664.738 Td /F1 9.8 Tf [( at least for C57BL/6\).)] TJ ET BT 26.250 645.333 Td /F1 9.8 Tf [(Although the differences in locomotion were in the expected direction, these differences were not statistically significant for the )] TJ ET BT 26.250 633.429 Td /F1 9.8 Tf [(R6/2 mice \()] TJ ET BT 76.092 633.429 Td /F5 9.8 Tf [(p)] TJ ET BT 81.513 633.429 Td /F1 9.8 Tf [(=.31\). On the other hand, same R6/2 mice were significantly more immobile than their wild-type controls. The )] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(dissociation between these two measures is possible since immobility and locomotion \(i.e., path length\) are not perfectly )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(complementary given all other alternative behaviors animals engage in \(e.g., grooming\). A more detailed characterization of )] TJ ET BT 26.250 597.714 Td /F1 9.8 Tf [(behavior via computer vision system in future studies can clarify the dynamics that might underlie this dissociation.)] TJ ET BT 26.250 578.310 Td /F1 9.8 Tf [(Since our computer vision application does not track the individual animal identities in a consistent manner \(see below for )] TJ ET BT 26.250 566.405 Td /F1 9.8 Tf [(details\), in order to compare genotypes in terms of computer vision indices, mice with the same genotype were housed in the )] TJ ET BT 26.250 554.500 Td /F1 9.8 Tf [(same cage. While, we could track the identity of a given mouse prior to and following corner exits \(based on time-stamped )] TJ ET BT 26.250 542.595 Td /F1 9.8 Tf [(transponder detections\), these periods of identity tracking were constrained by instances of occlusions, where identity swaps )] TJ ET BT 26.250 530.691 Td /F1 9.8 Tf [(are likely. Ongoing development of the computer vision application is focused on reliable individual tracking and social behavior. )] TJ ET BT 26.250 518.786 Td /F1 9.8 Tf [(Another advantage of genotype-based housing in our specific case was to avoid possible dominance of the WT mice over R6/2 )] TJ ET BT 26.250 506.881 Td /F1 9.8 Tf [(and BACHD mice minimizing their access to resources. This would have constituted an important confound during behavioral )] TJ ET BT 26.250 494.976 Td /F1 9.8 Tf [(phenotyping of these mice. Certain behavioral phenotypes of R6/2 and BACHD mice \(e.g., disrupted sleep patterns\) could also )] TJ ET BT 26.250 483.072 Td /F1 9.8 Tf [(directly interact with WT mouse behavior. Similar factors however could also constitute a disadvantage of genotype-based )] TJ ET BT 26.250 471.167 Td /F1 9.8 Tf [(housing schemes as they could inflate the degree of behavioral phenotype. For instance, disrupted sleep pattern of a given )] TJ ET BT 26.250 459.262 Td /F1 9.8 Tf [(mouse could also disrupt the sleep patterns of other mice in the same cage.)] TJ ET BT 26.250 439.857 Td /F1 9.8 Tf [(Group housing mice during testing might itself exert some disadvantages. One of the likely problems with this approach is that )] TJ ET BT 26.250 427.953 Td /F1 9.8 Tf [(group housing might induce social ranking dynamics, which might in turn influence behavior and task-related performances. In )] TJ ET BT 26.250 416.048 Td /F1 9.8 Tf [(this study, we did not specifically evaluate social ranking and thus could not capture these possible relations. On the other hand, )] TJ ET BT 26.250 404.143 Td /F1 9.8 Tf [(in combination with marking animals for individuation, analytical tools can be developed to evaluate social behavioral dynamics )] TJ ET BT 26.250 392.238 Td /F1 9.8 Tf [(based on computer vision application endpoints. For instance, huddling behavior and/or dispersal of animals in the home-cage )] TJ ET BT 26.250 380.334 Td /F1 9.8 Tf [(might provide important data regarding social behavior. Genotype-based housing or group-housing itself exerts a trade-off that )] TJ ET BT 26.250 368.429 Td /F1 9.8 Tf [(needs to be evaluated given what is known about the mouse lines to be tested.)] TJ ET BT 26.250 349.024 Td /F1 9.8 Tf [(Another important issue is the number of subjects to test in a given case. In our case, the number of subjects was chosen to )] TJ ET BT 26.250 337.119 Td /F1 9.8 Tf [(maximize the sample size \(as allowed by regulations\). Power analysis shows that group sizes of 3-11 mice for most of the )] TJ ET BT 26.250 325.215 Td /F1 9.8 Tf [(parameters measured in the R6/2 mice at 8 weeks of age \(namely: corner entries, licking count, percentage of repeats, )] TJ ET BT 26.250 313.310 Td /F1 9.8 Tf [(percentage of alternations, percent time immobile and percent time rearing-climbing\) are sufficient to detect a 50% improvement )] TJ ET BT 26.250 301.405 Td /F1 9.8 Tf [(of a potential therapy \(final number depends on the parameter measured; ?= 0.05, 80% power\). A similar sample size allows )] TJ ET BT 26.250 289.500 Td /F1 9.8 Tf [(the evaluation of potential therapies at 10 weeks of age R6/2 mice in the PhenoCube)] TJ ET BT 392.080 293.389 Td /F1 8.7 Tf [(TM)] TJ ET BT 404.594 289.500 Td /F1 9.8 Tf [( system when measuring percent )] TJ ET BT 26.250 277.596 Td /F1 9.8 Tf [(alternations, percent time immobile and percent time rearing-climbing. In the BACHD mice, power analysis indicates that group )] TJ ET BT 26.250 265.691 Td /F1 9.8 Tf [(sizes of 4-16 mice are sufficient to detect a 50% effect improvement at 36 weeks of age when measuring corner entries, )] TJ ET BT 26.250 253.786 Td /F1 9.8 Tf [(collected rewards, and locomotion. If the effect of the potential therapy is examined at 52 weeks of age, group sizes of 4-9 mice )] TJ ET BT 26.250 241.881 Td /F1 9.8 Tf [(are needed when measuring corner entries, collected rewards, locomotion, percent time immobile and percent time rearing-)] TJ ET BT 26.250 229.977 Td /F1 9.8 Tf [(climbing.)] TJ ET BT 26.250 210.572 Td /F1 9.8 Tf [(One of the important features of the data gathered from this study is the increased licking that could be interpreted as increased )] TJ ET BT 26.250 198.667 Td /F1 9.8 Tf [(thirst of both R6/2 and BACHD mice coupled with the use of water as reward. Increased licking phenotype might constitute a )] TJ ET BT 26.250 186.762 Td /F1 9.8 Tf [(confound for behavioral tasks that set the rules for access to water reward )] TJ ET 0.267 0.267 0.267 rg BT 348.146 188.270 Td /F4 8.7 Tf [(32)] TJ ET 0.271 0.267 0.267 rg BT 357.784 186.762 Td /F1 9.8 Tf [(. That have been said, the canonical interpretation )] TJ ET BT 26.250 174.858 Td /F1 9.8 Tf [(of higher thirst would be increased motivation for water reward, which if anything would be expected to lead to more well-)] TJ ET BT 26.250 162.953 Td /F1 9.8 Tf [(structured \(and possibly less disrupted\) goal-directed behaviors. On the other hand, several phenotypes such as the frequency )] TJ ET BT 26.250 151.048 Td /F1 9.8 Tf [(of corner visits might have been masked by higher motivation for water reward at least in the R6/2 mice.)] TJ ET BT 26.250 131.643 Td /F1 9.8 Tf [(Note that the aim of this study was not only to characterize the disease profiles of two different mutations, but instead to )] TJ ET BT 26.250 119.739 Td /F1 9.8 Tf [(demonstrate the utility of the PhenoCube)] TJ ET BT 202.930 123.627 Td /F1 8.7 Tf [(TM)] TJ ET BT 215.444 119.739 Td /F1 9.8 Tf [( in mouse models of HD. Further work can test different mouse models at the same )] TJ ET BT 26.250 107.834 Td /F1 9.8 Tf [(age \(e.g., BACHD vs. R6/2 mice at 8 weeks of age\) with identical experimental protocols to enable a direct comparison of )] TJ ET BT 26.250 95.929 Td /F1 9.8 Tf [(mouse models of HD using PhenoCube)] TJ ET BT 197.489 99.817 Td /F1 8.7 Tf [(TM)] TJ ET BT 210.004 95.929 Td /F1 9.8 Tf [( as a tool. Another interesting possibility for future studies would be mixing different )] TJ ET BT 26.250 84.024 Td /F1 9.8 Tf [(genotypes in a cage and comparing their performance to conditions where mice of the same genotype are housed together \(as )] TJ ET BT 26.250 72.120 Td /F1 9.8 Tf [(in this study\). This would enable the differentiation of disease progression in the presence and absence of mice with HD and )] TJ ET BT 26.250 60.215 Td /F1 9.8 Tf [(might reveal possibly differential social behavior dynamics compared to genotype-based housing scheme. Finally, defining the )] TJ ET BT 26.250 48.310 Td /F1 9.8 Tf [(accuracy of nose-poke based on randomly signaled water recesses would constitute a test of cued discrimination performance, )] TJ ET BT 26.250 36.405 Td /F1 9.8 Tf [(which can be compared to the spatial discrimination of recesses as in the case of this study. These two different strategies are )] TJ ET Q q 15.000 22.120 577.500 754.880 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(been observed in HD patients as assessed in different tasks )] TJ ET 0.267 0.267 0.267 rg BT 287.998 768.983 Td /F4 8.7 Tf [(26)] TJ ET 0.271 0.267 0.267 rg BT 297.636 771.364 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 300.045 768.983 Td /F4 8.7 Tf [(27)] TJ ET 0.271 0.267 0.267 rg BT 309.683 771.364 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 312.092 768.983 Td /F4 8.7 Tf [(28)] TJ ET 0.271 0.267 0.267 rg BT 321.729 771.364 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 324.139 768.983 Td /F4 8.7 Tf [(29)] TJ ET 0.271 0.267 0.267 rg BT 333.776 767.476 Td /F1 9.8 Tf [(.)] TJ ET BT 26.250 748.071 Td /F1 9.8 Tf [(We also evaluated the motor behavior of the R6/2 and BACHD mice using the computer vision software revealing that maybe )] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(R6/2 but clearly BACHD mice had shorter total path length, \(not significant for R6/2 mice\), reduced levels of rearing and )] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(climbing behavior, and higher immobility time. These results are consistent with previous reports of decreased locomotion )] TJ ET 0.267 0.267 0.267 rg BT 552.399 725.769 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 557.218 728.150 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 559.627 725.769 Td /F4 8.7 Tf [(33)] TJ ET 0.271 0.267 0.267 rg BT 569.264 724.262 Td /F1 9.8 Tf [(, )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(rearing-climbing )] TJ ET 0.267 0.267 0.267 rg BT 98.302 713.864 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 103.121 712.357 Td /F1 9.8 Tf [(, and imbalance and motor coordination problems )] TJ ET 0.267 0.267 0.267 rg BT 319.883 713.864 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 324.702 716.245 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 327.111 713.864 Td /F4 8.7 Tf [(20)] TJ ET 0.271 0.267 0.267 rg BT 336.749 712.357 Td /F1 9.8 Tf [( in R6/2 and BACHD mice. The performance of R6/2 )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(mice particularly paralleled the results of Rudenko et al )] TJ ET 0.267 0.267 0.267 rg BT 265.759 701.960 Td /F4 8.7 Tf [(25)] TJ ET 0.271 0.267 0.267 rg BT 275.396 700.452 Td /F1 9.8 Tf [( who tested these mice on open field )] TJ ET 0.267 0.267 0.267 rg BT 436.359 701.960 Td /F4 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 441.178 704.341 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 443.587 701.960 Td /F4 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 453.224 700.452 Td /F1 9.8 Tf [(. They observed significant )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(decrease in number of rearings as early as 6 weeks of age, however differences in distance traveled occurred reliably at 12 )] TJ ET BT 26.250 676.643 Td /F1 9.8 Tf [(weeks of age. Light cycle was found to affect many of our computer vision measures, which has also been reported in studies )] TJ ET BT 26.250 664.738 Td /F1 9.8 Tf [(that tested WT mice in open field \(e.g., )] TJ ET 0.267 0.267 0.267 rg BT 195.871 666.245 Td /F4 8.7 Tf [(34)] TJ ET 0.271 0.267 0.267 rg BT 205.508 668.626 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 207.917 666.245 Td /F4 8.7 Tf [(35)] TJ ET 0.271 0.267 0.267 rg BT 217.555 664.738 Td /F1 9.8 Tf [( at least for C57BL/6\).)] TJ ET BT 26.250 645.333 Td /F1 9.8 Tf [(Although the differences in locomotion were in the expected direction, these differences were not statistically significant for the )] TJ ET BT 26.250 633.429 Td /F1 9.8 Tf [(R6/2 mice \()] TJ ET BT 76.092 633.429 Td /F5 9.8 Tf [(p)] TJ ET BT 81.513 633.429 Td /F1 9.8 Tf [(=.31\). On the other hand, same R6/2 mice were significantly more immobile than their wild-type controls. The )] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(dissociation between these two measures is possible since immobility and locomotion \(i.e., path length\) are not perfectly )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(complementary given all other alternative behaviors animals engage in \(e.g., grooming\). A more detailed characterization of )] TJ ET BT 26.250 597.714 Td /F1 9.8 Tf [(behavior via computer vision system in future studies can clarify the dynamics that might underlie this dissociation.)] TJ ET BT 26.250 578.310 Td /F1 9.8 Tf [(Since our computer vision application does not track the individual animal identities in a consistent manner \(see below for )] TJ ET BT 26.250 566.405 Td /F1 9.8 Tf [(details\), in order to compare genotypes in terms of computer vision indices, mice with the same genotype were housed in the )] TJ ET BT 26.250 554.500 Td /F1 9.8 Tf [(same cage. While, we could track the identity of a given mouse prior to and following corner exits \(based on time-stamped )] TJ ET BT 26.250 542.595 Td /F1 9.8 Tf [(transponder detections\), these periods of identity tracking were constrained by instances of occlusions, where identity swaps )] TJ ET BT 26.250 530.691 Td /F1 9.8 Tf [(are likely. Ongoing development of the computer vision application is focused on reliable individual tracking and social behavior. )] TJ ET BT 26.250 518.786 Td /F1 9.8 Tf [(Another advantage of genotype-based housing in our specific case was to avoid possible dominance of the WT mice over R6/2 )] TJ ET BT 26.250 506.881 Td /F1 9.8 Tf [(and BACHD mice minimizing their access to resources. This would have constituted an important confound during behavioral )] TJ ET BT 26.250 494.976 Td /F1 9.8 Tf [(phenotyping of these mice. Certain behavioral phenotypes of R6/2 and BACHD mice \(e.g., disrupted sleep patterns\) could also )] TJ ET BT 26.250 483.072 Td /F1 9.8 Tf [(directly interact with WT mouse behavior. Similar factors however could also constitute a disadvantage of genotype-based )] TJ ET BT 26.250 471.167 Td /F1 9.8 Tf [(housing schemes as they could inflate the degree of behavioral phenotype. For instance, disrupted sleep pattern of a given )] TJ ET BT 26.250 459.262 Td /F1 9.8 Tf [(mouse could also disrupt the sleep patterns of other mice in the same cage.)] TJ ET BT 26.250 439.857 Td /F1 9.8 Tf [(Group housing mice during testing might itself exert some disadvantages. One of the likely problems with this approach is that )] TJ ET BT 26.250 427.953 Td /F1 9.8 Tf [(group housing might induce social ranking dynamics, which might in turn influence behavior and task-related performances. In )] TJ ET BT 26.250 416.048 Td /F1 9.8 Tf [(this study, we did not specifically evaluate social ranking and thus could not capture these possible relations. On the other hand, )] TJ ET BT 26.250 404.143 Td /F1 9.8 Tf [(in combination with marking animals for individuation, analytical tools can be developed to evaluate social behavioral dynamics )] TJ ET BT 26.250 392.238 Td /F1 9.8 Tf [(based on computer vision application endpoints. For instance, huddling behavior and/or dispersal of animals in the home-cage )] TJ ET BT 26.250 380.334 Td /F1 9.8 Tf [(might provide important data regarding social behavior. Genotype-based housing or group-housing itself exerts a trade-off that )] TJ ET BT 26.250 368.429 Td /F1 9.8 Tf [(needs to be evaluated given what is known about the mouse lines to be tested.)] TJ ET BT 26.250 349.024 Td /F1 9.8 Tf [(Another important issue is the number of subjects to test in a given case. In our case, the number of subjects was chosen to )] TJ ET BT 26.250 337.119 Td /F1 9.8 Tf [(maximize the sample size \(as allowed by regulations\). Power analysis shows that group sizes of 3-11 mice for most of the )] TJ ET BT 26.250 325.215 Td /F1 9.8 Tf [(parameters measured in the R6/2 mice at 8 weeks of age \(namely: corner entries, licking count, percentage of repeats, )] TJ ET BT 26.250 313.310 Td /F1 9.8 Tf [(percentage of alternations, percent time immobile and percent time rearing-climbing\) are sufficient to detect a 50% improvement )] TJ ET BT 26.250 301.405 Td /F1 9.8 Tf [(of a potential therapy \(final number depends on the parameter measured; ?= 0.05, 80% power\). A similar sample size allows )] TJ ET BT 26.250 289.500 Td /F1 9.8 Tf [(the evaluation of potential therapies at 10 weeks of age R6/2 mice in the PhenoCube)] TJ ET BT 392.080 293.389 Td /F1 8.7 Tf [(TM)] TJ ET BT 404.594 289.500 Td /F1 9.8 Tf [( system when measuring percent )] TJ ET BT 26.250 277.596 Td /F1 9.8 Tf [(alternations, percent time immobile and percent time rearing-climbing. In the BACHD mice, power analysis indicates that group )] TJ ET BT 26.250 265.691 Td /F1 9.8 Tf [(sizes of 4-16 mice are sufficient to detect a 50% effect improvement at 36 weeks of age when measuring corner entries, )] TJ ET BT 26.250 253.786 Td /F1 9.8 Tf [(collected rewards, and locomotion. If the effect of the potential therapy is examined at 52 weeks of age, group sizes of 4-9 mice )] TJ ET BT 26.250 241.881 Td /F1 9.8 Tf [(are needed when measuring corner entries, collected rewards, locomotion, percent time immobile and percent time rearing-)] TJ ET BT 26.250 229.977 Td /F1 9.8 Tf [(climbing.)] TJ ET BT 26.250 210.572 Td /F1 9.8 Tf [(One of the important features of the data gathered from this study is the increased licking that could be interpreted as increased )] TJ ET BT 26.250 198.667 Td /F1 9.8 Tf [(thirst of both R6/2 and BACHD mice coupled with the use of water as reward. Increased licking phenotype might constitute a )] TJ ET BT 26.250 186.762 Td /F1 9.8 Tf [(confound for behavioral tasks that set the rules for access to water reward )] TJ ET 0.267 0.267 0.267 rg BT 348.146 188.270 Td /F4 8.7 Tf [(32)] TJ ET 0.271 0.267 0.267 rg BT 357.784 186.762 Td /F1 9.8 Tf [(. That have been said, the canonical interpretation )] TJ ET BT 26.250 174.858 Td /F1 9.8 Tf [(of higher thirst would be increased motivation for water reward, which if anything would be expected to lead to more well-)] TJ ET BT 26.250 162.953 Td /F1 9.8 Tf [(structured \(and possibly less disrupted\) goal-directed behaviors. On the other hand, several phenotypes such as the frequency )] TJ ET BT 26.250 151.048 Td /F1 9.8 Tf [(of corner visits might have been masked by higher motivation for water reward at least in the R6/2 mice.)] TJ ET BT 26.250 131.643 Td /F1 9.8 Tf [(Note that the aim of this study was not only to characterize the disease profiles of two different mutations, but instead to )] TJ ET BT 26.250 119.739 Td /F1 9.8 Tf [(demonstrate the utility of the PhenoCube)] TJ ET BT 202.930 123.627 Td /F1 8.7 Tf [(TM)] TJ ET BT 215.444 119.739 Td /F1 9.8 Tf [( in mouse models of HD. Further work can test different mouse models at the same )] TJ ET BT 26.250 107.834 Td /F1 9.8 Tf [(age \(e.g., BACHD vs. R6/2 mice at 8 weeks of age\) with identical experimental protocols to enable a direct comparison of )] TJ ET BT 26.250 95.929 Td /F1 9.8 Tf [(mouse models of HD using PhenoCube)] TJ ET BT 197.489 99.817 Td /F1 8.7 Tf [(TM)] TJ ET BT 210.004 95.929 Td /F1 9.8 Tf [( as a tool. Another interesting possibility for future studies would be mixing different )] TJ ET BT 26.250 84.024 Td /F1 9.8 Tf [(genotypes in a cage and comparing their performance to conditions where mice of the same genotype are housed together \(as )] TJ ET BT 26.250 72.120 Td /F1 9.8 Tf [(in this study\). This would enable the differentiation of disease progression in the presence and absence of mice with HD and )] TJ ET BT 26.250 60.215 Td /F1 9.8 Tf [(might reveal possibly differential social behavior dynamics compared to genotype-based housing scheme. Finally, defining the )] TJ ET BT 26.250 48.310 Td /F1 9.8 Tf [(accuracy of nose-poke based on randomly signaled water recesses would constitute a test of cued discrimination performance, )] TJ ET BT 26.250 36.405 Td /F1 9.8 Tf [(which can be compared to the spatial discrimination of recesses as in the case of this study. 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Morton, 2009\).)] TJ ET BT 26.250 748.071 Td /F1 9.8 Tf [(Overall, these results in HD mouse models characterize PhenoCube)] TJ ET BT 321.022 751.960 Td /F1 8.7 Tf [(TM)] TJ ET BT 333.536 748.071 Td /F1 9.8 Tf [( as an ecologically valid, sensitive, and efficient system )] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(in detecting and tracking the course of the phenotype progression concurrently across multiple behavioral measures. As )] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(practice effects are possible in longitudinal studies, confirmation of disease progression should be obtained using cross-)] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(sectional designs. Importantly, PhenoCube)] TJ ET BT 211.588 716.245 Td /F1 8.7 Tf [(TM)] TJ ET BT 224.102 712.357 Td /F1 9.8 Tf [( allows assessment of behavior in non-stressful conditions and of grouped mice )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(without any handling of the animals. Further refinement of the computer vision for assessment of social behavior will allow the )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(study of complex interactions in heterogeneous groups of subjects, a topic of particular interest in preclinical work with models )] TJ ET BT 26.250 676.643 Td /F1 9.8 Tf [(of schizophrenia and autism. In addition, the automated computer vision scoring provides measures of behavior 30 times/s, 24 )] TJ ET BT 26.250 664.738 Td /F1 9.8 Tf [(h/day, for up to 6 consecutive days, giving unprecedented access to the secret life of mice \(and rats in the near future\), and )] TJ ET BT 26.250 652.833 Td /F1 9.8 Tf [(offering the potential to discover unexpected aspects of their individual and group behavior. Critically, the ability to conduct )] TJ ET BT 26.250 640.929 Td /F1 9.8 Tf [(longitudinal studies is necessary for not only validation of disease models but also for the initial screening of potential )] TJ ET BT 26.250 629.024 Td /F1 9.8 Tf [(symptomatic treatments and therapeutic interventions. Results gathered from PhenoCube)] TJ ET BT 413.169 632.912 Td /F1 8.7 Tf [(TM)] TJ ET BT 425.684 629.024 Td /F1 9.8 Tf [( can be used to guide further )] TJ ET BT 26.250 617.119 Td /F1 9.8 Tf [(efforts using traditional assays. Thus, this new platform can be beneficial in eliminating the bottleneck in target validation and )] TJ ET BT 26.250 605.214 Td /F1 9.8 Tf [(drug discovery.)] TJ ET BT 26.250 568.612 Td /F4 12.0 Tf [(References)] TJ ET BT 26.250 541.158 Td /F1 9.8 Tf [(1.)] TJ ET BT 38.132 541.158 Td /F1 9.8 Tf [(Maggi S, Luciana G, Heise I, Nieus T, Balc? F, Wells S, Tocchini-Valentini GP, Mandillo S, Nolan PM, Tucci V. A cross-)] TJ ET BT 26.250 529.253 Td /F1 9.8 Tf [(laboratory investigation of timing endophenotypes in mouse behavior. Timing & Time Perception. in press.)] TJ ET BT 26.250 509.848 Td /F1 9.8 Tf [(2.)] TJ ET BT 38.132 509.848 Td /F1 9.8 Tf [(Lipp, H.-P. \(2005\). High-throughput and Automated Behavioural Screening of Normal and Genetically Modified Mice, )] TJ ET BT 26.250 497.943 Td /F1 9.8 Tf [(Business Briefing: Future Drug Discovery.)] TJ ET BT 26.250 478.539 Td /F1 9.8 Tf [(3.)] TJ ET BT 38.132 478.539 Td /F1 9.8 Tf [(Gallistel CR, King AP, Daniel AM, Freestone D, Papachristos EB, Balci F, Kheifets A, Zhang J, Su X, Schiff G, Kourtev H. )] TJ ET BT 26.250 466.634 Td /F1 9.8 Tf [(Screening for Learning and Memory Mutations: A New Approach. Xin Li Xue Bao. 2010 Jan 30;42\(1\):138-158. PubMed )] TJ ET BT 26.250 454.729 Td /F1 9.8 Tf [(PMID:20352069.)] TJ ET BT 26.250 435.324 Td /F1 9.8 Tf [(4.)] TJ ET BT 38.132 435.324 Td /F1 9.8 Tf [(Gray M, Shirasaki DI, Cepeda C, Andr VM, Wilburn B, Lu XH, Tao J, Yamazaki I, Li SH, Sun YE, Li XJ, Levine MS, Yang )] TJ ET BT 26.250 423.420 Td /F1 9.8 Tf [(XW. Full-length human mutant huntingtin with a stable polyglutamine repeat can elicit progressive and selective )] TJ ET BT 26.250 411.515 Td /F1 9.8 Tf [(neuropathogenesis in BACHD mice. J Neurosci. 2008 Jun 11;28\(24\):6182-95. PubMed PMID:18550760.)] TJ ET BT 26.250 392.110 Td /F1 9.8 Tf [(5.)] TJ ET BT 38.132 392.110 Td /F1 9.8 Tf [(Mangiarini L, Sathasivam K, Mahal A, Mott R, Seller M, Bates GP. Instability of highly expanded CAG repeats in mice )] TJ ET BT 26.250 380.205 Td /F1 9.8 Tf [(transgenic for the Huntington's disease mutation. Nat Genet. 1997 Feb;15\(2\):197-200. PubMed PMID:9020849.)] TJ ET BT 26.250 360.801 Td /F1 9.8 Tf [(6.)] TJ ET BT 38.132 360.801 Td /F1 9.8 Tf [(Ciamei A, Morton AJ. Rigidity in social and emotional memory in the R6/2 mouse model of Huntington's disease. Neurobiol )] TJ ET BT 26.250 348.896 Td /F1 9.8 Tf [(Learn Mem. 2008 May;89\(4\):533-44. PubMed PMID:18069020.)] TJ ET BT 26.250 329.491 Td /F1 9.8 Tf [(7.)] TJ ET BT 38.132 329.491 Td /F1 9.8 Tf [(Ciamei A, Morton AJ. Rigidity in social and emotional memory in the R6/2 mouse model of Huntington's disease. Neurobiol )] TJ ET BT 26.250 317.586 Td /F1 9.8 Tf [(Learn Mem. 2008 May;89\(4\):533-44. PubMed PMID:18069020.)] TJ ET BT 26.250 298.182 Td /F1 9.8 Tf [(8.)] TJ ET BT 38.132 298.182 Td /F1 9.8 Tf [(Lione LA, Carter RJ, Hunt MJ, Bates GP, Morton AJ, Dunnett SB. Selective discrimination learning impairments in mice )] TJ ET BT 26.250 286.277 Td /F1 9.8 Tf [(expressing the human Huntington's disease mutation. J Neurosci. 1999 Dec 1;19\(23\):10428-37. PubMed PMID:10575040.)] TJ ET BT 26.250 266.872 Td /F1 9.8 Tf [(9.)] TJ ET BT 38.132 266.872 Td /F1 9.8 Tf [(Menalled L, El-Khodor BF, Patry M, Surez-Farias M, Orenstein SJ, Zahasky B, Leahy C, Wheeler V, Yang XW, )] TJ ET BT 26.250 254.967 Td /F1 9.8 Tf [(MacDonald M, Morton AJ, Bates G, Leeds J, Park L, Howland D, Signer E, Tobin A, Brunner D. Systematic behavioral )] TJ ET BT 26.250 243.063 Td /F1 9.8 Tf [(evaluation of Huntington's disease transgenic and knock-in mouse models. Neurobiol Dis. 2009 Sep;35\(3\):319-36. PubMed )] TJ ET BT 26.250 231.158 Td /F1 9.8 Tf [(PMID:19464370.)] TJ ET BT 26.250 211.753 Td /F1 9.8 Tf [(10.)] TJ ET BT 43.553 211.753 Td /F1 9.8 Tf [(Morton AJ, Skillings E, Bussey TJ, Saksida LM. Measuring cognitive deficits in disabled mice using an automated )] TJ ET BT 26.250 199.848 Td /F1 9.8 Tf [(interactive touchscreen system. Nat Methods. 2006 Oct;3\(10\):767. PubMed PMID:16990806.)] TJ ET BT 26.250 180.444 Td /F1 9.8 Tf [(11.)] TJ ET BT 43.553 180.444 Td /F1 9.8 Tf [(Pallier PN, Maywood ES, Zheng Z, Chesham JE, Inyushkin AN, Dyball R, Hastings MH, Morton AJ. Pharmacological )] TJ ET BT 26.250 168.539 Td /F1 9.8 Tf [(imposition of sleep slows cognitive decline and reverses dysregulation of circadian gene expression in a transgenic mouse )] TJ ET BT 26.250 156.634 Td /F1 9.8 Tf [(model of Huntington's disease. J Neurosci. 2007 Jul 18;27\(29\):7869-78. PubMed PMID:17634381.)] TJ ET BT 26.250 137.229 Td /F1 9.8 Tf [(12.)] TJ ET BT 43.553 137.229 Td /F1 9.8 Tf [(Carter RJ, Lione LA, Humby T, Mangiarini L, Mahal A, Bates GP, Dunnett SB, Morton AJ. Characterization of progressive )] TJ ET BT 26.250 125.325 Td /F1 9.8 Tf [(motor deficits in mice transgenic for the human Huntington's disease mutation. J Neurosci. 1999 Apr 15;19\(8\):3248-57. PubMed )] TJ ET BT 26.250 113.420 Td /F1 9.8 Tf [(PMID:10191337.)] TJ ET BT 26.250 94.015 Td /F1 9.8 Tf [(13.)] TJ ET BT 43.553 94.015 Td /F1 9.8 Tf [(Morton AJ, Wood NI, Hastings MH, Hurelbrink C, Barker RA, Maywood ES. Disintegration of the sleep-wake cycle and )] TJ ET BT 26.250 82.110 Td /F1 9.8 Tf [(circadian timing in Huntington's disease. J Neurosci. 2005 Jan 5;25\(1\):157-63. PubMed PMID:15634777.)] TJ ET BT 26.250 62.706 Td /F1 9.8 Tf [(14.)] TJ ET BT 43.553 62.706 Td /F1 9.8 Tf [(Oakeshott S, Port RG, Cummins-Sutphen J, Watson-Johnson J, Ramboz S, Park L, Howland D, Brunner D. HD mouse )] TJ ET BT 26.250 50.801 Td /F1 9.8 Tf [(models reveal clear deficits in learning to perform a simple instrumental response. PLoS Curr. 2011a Nov 30;3:RRN1282. )] TJ ET Q q 15.000 36.515 577.500 740.485 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(known to rely on different mechanisms \(e.g., Ciamei & Morton, 2009\).)] TJ ET BT 26.250 748.071 Td /F1 9.8 Tf [(Overall, these results in HD mouse models characterize PhenoCube)] TJ ET BT 321.022 751.960 Td /F1 8.7 Tf [(TM)] TJ ET BT 333.536 748.071 Td /F1 9.8 Tf [( as an ecologically valid, sensitive, and efficient system )] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(in detecting and tracking the course of the phenotype progression concurrently across multiple behavioral measures. As )] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(practice effects are possible in longitudinal studies, confirmation of disease progression should be obtained using cross-)] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(sectional designs. Importantly, PhenoCube)] TJ ET BT 211.588 716.245 Td /F1 8.7 Tf [(TM)] TJ ET BT 224.102 712.357 Td /F1 9.8 Tf [( allows assessment of behavior in non-stressful conditions and of grouped mice )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(without any handling of the animals. Further refinement of the computer vision for assessment of social behavior will allow the )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(study of complex interactions in heterogeneous groups of subjects, a topic of particular interest in preclinical work with models )] TJ ET BT 26.250 676.643 Td /F1 9.8 Tf [(of schizophrenia and autism. In addition, the automated computer vision scoring provides measures of behavior 30 times/s, 24 )] TJ ET BT 26.250 664.738 Td /F1 9.8 Tf [(h/day, for up to 6 consecutive days, giving unprecedented access to the secret life of mice \(and rats in the near future\), and )] TJ ET BT 26.250 652.833 Td /F1 9.8 Tf [(offering the potential to discover unexpected aspects of their individual and group behavior. Critically, the ability to conduct )] TJ ET BT 26.250 640.929 Td /F1 9.8 Tf [(longitudinal studies is necessary for not only validation of disease models but also for the initial screening of potential )] TJ ET BT 26.250 629.024 Td /F1 9.8 Tf [(symptomatic treatments and therapeutic interventions. Results gathered from PhenoCube)] TJ ET BT 413.169 632.912 Td /F1 8.7 Tf [(TM)] TJ ET BT 425.684 629.024 Td /F1 9.8 Tf [( can be used to guide further )] TJ ET BT 26.250 617.119 Td /F1 9.8 Tf [(efforts using traditional assays. Thus, this new platform can be beneficial in eliminating the bottleneck in target validation and )] TJ ET BT 26.250 605.214 Td /F1 9.8 Tf [(drug discovery.)] TJ ET BT 26.250 568.612 Td /F4 12.0 Tf [(References)] TJ ET BT 26.250 541.158 Td /F1 9.8 Tf [(1.)] TJ ET BT 38.132 541.158 Td /F1 9.8 Tf [(Maggi S, Luciana G, Heise I, Nieus T, Balc? F, Wells S, Tocchini-Valentini GP, Mandillo S, Nolan PM, Tucci V. A cross-)] TJ ET BT 26.250 529.253 Td /F1 9.8 Tf [(laboratory investigation of timing endophenotypes in mouse behavior. Timing & Time Perception. in press.)] TJ ET BT 26.250 509.848 Td /F1 9.8 Tf [(2.)] TJ ET BT 38.132 509.848 Td /F1 9.8 Tf [(Lipp, H.-P. \(2005\). High-throughput and Automated Behavioural Screening of Normal and Genetically Modified Mice, )] TJ ET BT 26.250 497.943 Td /F1 9.8 Tf [(Business Briefing: Future Drug Discovery.)] TJ ET BT 26.250 478.539 Td /F1 9.8 Tf [(3.)] TJ ET BT 38.132 478.539 Td /F1 9.8 Tf [(Gallistel CR, King AP, Daniel AM, Freestone D, Papachristos EB, Balci F, Kheifets A, Zhang J, Su X, Schiff G, Kourtev H. )] TJ ET BT 26.250 466.634 Td /F1 9.8 Tf [(Screening for Learning and Memory Mutations: A New Approach. Xin Li Xue Bao. 2010 Jan 30;42\(1\):138-158. PubMed )] TJ ET BT 26.250 454.729 Td /F1 9.8 Tf [(PMID:20352069.)] TJ ET BT 26.250 435.324 Td /F1 9.8 Tf [(4.)] TJ ET BT 38.132 435.324 Td /F1 9.8 Tf [(Gray M, Shirasaki DI, Cepeda C, Andr VM, Wilburn B, Lu XH, Tao J, Yamazaki I, Li SH, Sun YE, Li XJ, Levine MS, Yang )] TJ ET BT 26.250 423.420 Td /F1 9.8 Tf [(XW. Full-length human mutant huntingtin with a stable polyglutamine repeat can elicit progressive and selective )] TJ ET BT 26.250 411.515 Td /F1 9.8 Tf [(neuropathogenesis in BACHD mice. J Neurosci. 2008 Jun 11;28\(24\):6182-95. PubMed PMID:18550760.)] TJ ET BT 26.250 392.110 Td /F1 9.8 Tf [(5.)] TJ ET BT 38.132 392.110 Td /F1 9.8 Tf [(Mangiarini L, Sathasivam K, Mahal A, Mott R, Seller M, Bates GP. Instability of highly expanded CAG repeats in mice )] TJ ET BT 26.250 380.205 Td /F1 9.8 Tf [(transgenic for the Huntington's disease mutation. Nat Genet. 1997 Feb;15\(2\):197-200. PubMed PMID:9020849.)] TJ ET BT 26.250 360.801 Td /F1 9.8 Tf [(6.)] TJ ET BT 38.132 360.801 Td /F1 9.8 Tf [(Ciamei A, Morton AJ. Rigidity in social and emotional memory in the R6/2 mouse model of Huntington's disease. Neurobiol )] TJ ET BT 26.250 348.896 Td /F1 9.8 Tf [(Learn Mem. 2008 May;89\(4\):533-44. PubMed PMID:18069020.)] TJ ET BT 26.250 329.491 Td /F1 9.8 Tf [(7.)] TJ ET BT 38.132 329.491 Td /F1 9.8 Tf [(Ciamei A, Morton AJ. Rigidity in social and emotional memory in the R6/2 mouse model of Huntington's disease. Neurobiol )] TJ ET BT 26.250 317.586 Td /F1 9.8 Tf [(Learn Mem. 2008 May;89\(4\):533-44. PubMed PMID:18069020.)] TJ ET BT 26.250 298.182 Td /F1 9.8 Tf [(8.)] TJ ET BT 38.132 298.182 Td /F1 9.8 Tf [(Lione LA, Carter RJ, Hunt MJ, Bates GP, Morton AJ, Dunnett SB. Selective discrimination learning impairments in mice )] TJ ET BT 26.250 286.277 Td /F1 9.8 Tf [(expressing the human Huntington's disease mutation. J Neurosci. 1999 Dec 1;19\(23\):10428-37. PubMed PMID:10575040.)] TJ ET BT 26.250 266.872 Td /F1 9.8 Tf [(9.)] TJ ET BT 38.132 266.872 Td /F1 9.8 Tf [(Menalled L, El-Khodor BF, Patry M, Surez-Farias M, Orenstein SJ, Zahasky B, Leahy C, Wheeler V, Yang XW, )] TJ ET BT 26.250 254.967 Td /F1 9.8 Tf [(MacDonald M, Morton AJ, Bates G, Leeds J, Park L, Howland D, Signer E, Tobin A, Brunner D. Systematic behavioral )] TJ ET BT 26.250 243.063 Td /F1 9.8 Tf [(evaluation of Huntington's disease transgenic and knock-in mouse models. Neurobiol Dis. 2009 Sep;35\(3\):319-36. PubMed )] TJ ET BT 26.250 231.158 Td /F1 9.8 Tf [(PMID:19464370.)] TJ ET BT 26.250 211.753 Td /F1 9.8 Tf [(10.)] TJ ET BT 43.553 211.753 Td /F1 9.8 Tf [(Morton AJ, Skillings E, Bussey TJ, Saksida LM. Measuring cognitive deficits in disabled mice using an automated )] TJ ET BT 26.250 199.848 Td /F1 9.8 Tf [(interactive touchscreen system. Nat Methods. 2006 Oct;3\(10\):767. PubMed PMID:16990806.)] TJ ET BT 26.250 180.444 Td /F1 9.8 Tf [(11.)] TJ ET BT 43.553 180.444 Td /F1 9.8 Tf [(Pallier PN, Maywood ES, Zheng Z, Chesham JE, Inyushkin AN, Dyball R, Hastings MH, Morton AJ. Pharmacological )] TJ ET BT 26.250 168.539 Td /F1 9.8 Tf [(imposition of sleep slows cognitive decline and reverses dysregulation of circadian gene expression in a transgenic mouse )] TJ ET BT 26.250 156.634 Td /F1 9.8 Tf [(model of Huntington's disease. J Neurosci. 2007 Jul 18;27\(29\):7869-78. PubMed PMID:17634381.)] TJ ET BT 26.250 137.229 Td /F1 9.8 Tf [(12.)] TJ ET BT 43.553 137.229 Td /F1 9.8 Tf [(Carter RJ, Lione LA, Humby T, Mangiarini L, Mahal A, Bates GP, Dunnett SB, Morton AJ. Characterization of progressive )] TJ ET BT 26.250 125.325 Td /F1 9.8 Tf [(motor deficits in mice transgenic for the human Huntington's disease mutation. J Neurosci. 1999 Apr 15;19\(8\):3248-57. PubMed )] TJ ET BT 26.250 113.420 Td /F1 9.8 Tf [(PMID:10191337.)] TJ ET BT 26.250 94.015 Td /F1 9.8 Tf [(13.)] TJ ET BT 43.553 94.015 Td /F1 9.8 Tf [(Morton AJ, Wood NI, Hastings MH, Hurelbrink C, Barker RA, Maywood ES. Disintegration of the sleep-wake cycle and )] TJ ET BT 26.250 82.110 Td /F1 9.8 Tf [(circadian timing in Huntington's disease. J Neurosci. 2005 Jan 5;25\(1\):157-63. PubMed PMID:15634777.)] TJ ET BT 26.250 62.706 Td /F1 9.8 Tf [(14.)] TJ ET BT 43.553 62.706 Td /F1 9.8 Tf [(Oakeshott S, Port RG, Cummins-Sutphen J, Watson-Johnson J, Ramboz S, Park L, Howland D, Brunner D. HD mouse )] TJ ET BT 26.250 50.801 Td /F1 9.8 Tf [(models reveal clear deficits in learning to perform a simple instrumental response. PLoS Curr. 2011a Nov 30;3:RRN1282. )] TJ ET Q q 15.000 36.515 577.500 740.485 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(known to rely on different mechanisms \(e.g., Ciamei & Morton, 2009\).)] TJ ET BT 26.250 748.071 Td /F1 9.8 Tf [(Overall, these results in HD mouse models characterize PhenoCube)] TJ ET BT 321.022 751.960 Td /F1 8.7 Tf [(TM)] TJ ET BT 333.536 748.071 Td /F1 9.8 Tf [( as an ecologically valid, sensitive, and efficient system )] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(in detecting and tracking the course of the phenotype progression concurrently across multiple behavioral measures. As )] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(practice effects are possible in longitudinal studies, confirmation of disease progression should be obtained using cross-)] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(sectional designs. Importantly, PhenoCube)] TJ ET BT 211.588 716.245 Td /F1 8.7 Tf [(TM)] TJ ET BT 224.102 712.357 Td /F1 9.8 Tf [( allows assessment of behavior in non-stressful conditions and of grouped mice )] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(without any handling of the animals. Further refinement of the computer vision for assessment of social behavior will allow the )] TJ ET BT 26.250 688.548 Td /F1 9.8 Tf [(study of complex interactions in heterogeneous groups of subjects, a topic of particular interest in preclinical work with models )] TJ ET BT 26.250 676.643 Td /F1 9.8 Tf [(of schizophrenia and autism. In addition, the automated computer vision scoring provides measures of behavior 30 times/s, 24 )] TJ ET BT 26.250 664.738 Td /F1 9.8 Tf [(h/day, for up to 6 consecutive days, giving unprecedented access to the secret life of mice \(and rats in the near future\), and )] TJ ET BT 26.250 652.833 Td /F1 9.8 Tf [(offering the potential to discover unexpected aspects of their individual and group behavior. Critically, the ability to conduct )] TJ ET BT 26.250 640.929 Td /F1 9.8 Tf [(longitudinal studies is necessary for not only validation of disease models but also for the initial screening of potential )] TJ ET BT 26.250 629.024 Td /F1 9.8 Tf [(symptomatic treatments and therapeutic interventions. Results gathered from PhenoCube)] TJ ET BT 413.169 632.912 Td /F1 8.7 Tf [(TM)] TJ ET BT 425.684 629.024 Td /F1 9.8 Tf [( can be used to guide further )] TJ ET BT 26.250 617.119 Td /F1 9.8 Tf [(efforts using traditional assays. Thus, this new platform can be beneficial in eliminating the bottleneck in target validation and )] TJ ET BT 26.250 605.214 Td /F1 9.8 Tf [(drug discovery.)] TJ ET BT 26.250 568.612 Td /F4 12.0 Tf [(References)] TJ ET BT 26.250 541.158 Td /F1 9.8 Tf [(1.)] TJ ET BT 38.132 541.158 Td /F1 9.8 Tf [(Maggi S, Luciana G, Heise I, Nieus T, Balc? F, Wells S, Tocchini-Valentini GP, Mandillo S, Nolan PM, Tucci V. A cross-)] TJ ET BT 26.250 529.253 Td /F1 9.8 Tf [(laboratory investigation of timing endophenotypes in mouse behavior. Timing & Time Perception. in press.)] TJ ET BT 26.250 509.848 Td /F1 9.8 Tf [(2.)] TJ ET BT 38.132 509.848 Td /F1 9.8 Tf [(Lipp, H.-P. \(2005\). High-throughput and Automated Behavioural Screening of Normal and Genetically Modified Mice, )] TJ ET BT 26.250 497.943 Td /F1 9.8 Tf [(Business Briefing: Future Drug Discovery.)] TJ ET BT 26.250 478.539 Td /F1 9.8 Tf [(3.)] TJ ET BT 38.132 478.539 Td /F1 9.8 Tf [(Gallistel CR, King AP, Daniel AM, Freestone D, Papachristos EB, Balci F, Kheifets A, Zhang J, Su X, Schiff G, Kourtev H. )] TJ ET BT 26.250 466.634 Td /F1 9.8 Tf [(Screening for Learning and Memory Mutations: A New Approach. Xin Li Xue Bao. 2010 Jan 30;42\(1\):138-158. PubMed )] TJ ET BT 26.250 454.729 Td /F1 9.8 Tf [(PMID:20352069.)] TJ ET BT 26.250 435.324 Td /F1 9.8 Tf [(4.)] TJ ET BT 38.132 435.324 Td /F1 9.8 Tf [(Gray M, Shirasaki DI, Cepeda C, Andr VM, Wilburn B, Lu XH, Tao J, Yamazaki I, Li SH, Sun YE, Li XJ, Levine MS, Yang )] TJ ET BT 26.250 423.420 Td /F1 9.8 Tf [(XW. Full-length human mutant huntingtin with a stable polyglutamine repeat can elicit progressive and selective )] TJ ET BT 26.250 411.515 Td /F1 9.8 Tf [(neuropathogenesis in BACHD mice. J Neurosci. 2008 Jun 11;28\(24\):6182-95. PubMed PMID:18550760.)] TJ ET BT 26.250 392.110 Td /F1 9.8 Tf [(5.)] TJ ET BT 38.132 392.110 Td /F1 9.8 Tf [(Mangiarini L, Sathasivam K, Mahal A, Mott R, Seller M, Bates GP. Instability of highly expanded CAG repeats in mice )] TJ ET BT 26.250 380.205 Td /F1 9.8 Tf [(transgenic for the Huntington's disease mutation. Nat Genet. 1997 Feb;15\(2\):197-200. PubMed PMID:9020849.)] TJ ET BT 26.250 360.801 Td /F1 9.8 Tf [(6.)] TJ ET BT 38.132 360.801 Td /F1 9.8 Tf [(Ciamei A, Morton AJ. Rigidity in social and emotional memory in the R6/2 mouse model of Huntington's disease. Neurobiol )] TJ ET BT 26.250 348.896 Td /F1 9.8 Tf [(Learn Mem. 2008 May;89\(4\):533-44. PubMed PMID:18069020.)] TJ ET BT 26.250 329.491 Td /F1 9.8 Tf [(7.)] TJ ET BT 38.132 329.491 Td /F1 9.8 Tf [(Ciamei A, Morton AJ. Rigidity in social and emotional memory in the R6/2 mouse model of Huntington's disease. Neurobiol )] TJ ET BT 26.250 317.586 Td /F1 9.8 Tf [(Learn Mem. 2008 May;89\(4\):533-44. PubMed PMID:18069020.)] TJ ET BT 26.250 298.182 Td /F1 9.8 Tf [(8.)] TJ ET BT 38.132 298.182 Td /F1 9.8 Tf [(Lione LA, Carter RJ, Hunt MJ, Bates GP, Morton AJ, Dunnett SB. Selective discrimination learning impairments in mice )] TJ ET BT 26.250 286.277 Td /F1 9.8 Tf [(expressing the human Huntington's disease mutation. J Neurosci. 1999 Dec 1;19\(23\):10428-37. PubMed PMID:10575040.)] TJ ET BT 26.250 266.872 Td /F1 9.8 Tf [(9.)] TJ ET BT 38.132 266.872 Td /F1 9.8 Tf [(Menalled L, El-Khodor BF, Patry M, Surez-Farias M, Orenstein SJ, Zahasky B, Leahy C, Wheeler V, Yang XW, )] TJ ET BT 26.250 254.967 Td /F1 9.8 Tf [(MacDonald M, Morton AJ, Bates G, Leeds J, Park L, Howland D, Signer E, Tobin A, Brunner D. Systematic behavioral )] TJ ET BT 26.250 243.063 Td /F1 9.8 Tf [(evaluation of Huntington's disease transgenic and knock-in mouse models. Neurobiol Dis. 2009 Sep;35\(3\):319-36. PubMed )] TJ ET BT 26.250 231.158 Td /F1 9.8 Tf [(PMID:19464370.)] TJ ET BT 26.250 211.753 Td /F1 9.8 Tf [(10.)] TJ ET BT 43.553 211.753 Td /F1 9.8 Tf [(Morton AJ, Skillings E, Bussey TJ, Saksida LM. Measuring cognitive deficits in disabled mice using an automated )] TJ ET BT 26.250 199.848 Td /F1 9.8 Tf [(interactive touchscreen system. Nat Methods. 2006 Oct;3\(10\):767. PubMed PMID:16990806.)] TJ ET BT 26.250 180.444 Td /F1 9.8 Tf [(11.)] TJ ET BT 43.553 180.444 Td /F1 9.8 Tf [(Pallier PN, Maywood ES, Zheng Z, Chesham JE, Inyushkin AN, Dyball R, Hastings MH, Morton AJ. Pharmacological )] TJ ET BT 26.250 168.539 Td /F1 9.8 Tf [(imposition of sleep slows cognitive decline and reverses dysregulation of circadian gene expression in a transgenic mouse )] TJ ET BT 26.250 156.634 Td /F1 9.8 Tf [(model of Huntington's disease. J Neurosci. 2007 Jul 18;27\(29\):7869-78. PubMed PMID:17634381.)] TJ ET BT 26.250 137.229 Td /F1 9.8 Tf [(12.)] TJ ET BT 43.553 137.229 Td /F1 9.8 Tf [(Carter RJ, Lione LA, Humby T, Mangiarini L, Mahal A, Bates GP, Dunnett SB, Morton AJ. Characterization of progressive )] TJ ET BT 26.250 125.325 Td /F1 9.8 Tf [(motor deficits in mice transgenic for the human Huntington's disease mutation. J Neurosci. 1999 Apr 15;19\(8\):3248-57. PubMed )] TJ ET BT 26.250 113.420 Td /F1 9.8 Tf [(PMID:10191337.)] TJ ET BT 26.250 94.015 Td /F1 9.8 Tf [(13.)] TJ ET BT 43.553 94.015 Td /F1 9.8 Tf [(Morton AJ, Wood NI, Hastings MH, Hurelbrink C, Barker RA, Maywood ES. Disintegration of the sleep-wake cycle and )] TJ ET BT 26.250 82.110 Td /F1 9.8 Tf [(circadian timing in Huntington's disease. J Neurosci. 2005 Jan 5;25\(1\):157-63. PubMed PMID:15634777.)] TJ ET BT 26.250 62.706 Td /F1 9.8 Tf [(14.)] TJ ET BT 43.553 62.706 Td /F1 9.8 Tf [(Oakeshott S, Port RG, Cummins-Sutphen J, Watson-Johnson J, Ramboz S, Park L, Howland D, Brunner D. HD mouse )] TJ ET BT 26.250 50.801 Td /F1 9.8 Tf [(models reveal clear deficits in learning to perform a simple instrumental response. PLoS Curr. 2011a Nov 30;3:RRN1282. )] TJ ET Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(12)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Huntington Disease)] TJ ET Q endstream endobj 585 0 obj << /Type /Page /Parent 3 0 R /Contents 586 0 R >> endobj 586 0 obj << /Length 23570 >> stream 0.271 0.267 0.267 rg q 15.000 48.072 577.500 728.928 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(PubMed PMID:22512000.)] TJ ET BT 26.250 748.071 Td /F1 9.8 Tf [(15.)] TJ ET BT 43.553 748.071 Td /F1 9.8 Tf [(Balci F, Day M, Rooney A, Brunner D. Disrupted temporal control in the R6/2 mouse model of Huntington's disease. Behav )] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(Neurosci. 2009 Dec;123\(6\):1353-8. PubMed PMID:20001119.)] TJ ET BT 26.250 716.762 Td /F1 9.8 Tf [(16.)] TJ ET BT 43.553 716.762 Td /F1 9.8 Tf [(Grote HE, Bull ND, Howard ML, van Dellen A, Blakemore C, Bartlett PF, Hannan AJ. Cognitive disorders and neurogenesis )] TJ ET BT 26.250 704.857 Td /F1 9.8 Tf [(deficits in Huntington's disease mice are rescued by fluoxetine. Eur J Neurosci. 2005 Oct;22\(8\):2081-8. PubMed )] TJ ET BT 26.250 692.952 Td /F1 9.8 Tf [(PMID:16262645.)] TJ ET BT 26.250 673.548 Td /F1 9.8 Tf [(17.)] TJ ET BT 43.553 673.548 Td /F1 9.8 Tf [(Murphy KP, Carter RJ, Lione LA, Mangiarini L, Mahal A, Bates GP, Dunnett SB, Morton AJ. Abnormal synaptic plasticity )] TJ ET BT 26.250 661.643 Td /F1 9.8 Tf [(and impaired spatial cognition in mice transgenic for exon 1 of the human Huntington's disease mutation. J Neurosci. 2000 Jul )] TJ ET BT 26.250 649.738 Td /F1 9.8 Tf [(1;20\(13\):5115-23. PubMed PMID:10864968.)] TJ ET BT 26.250 630.333 Td /F1 9.8 Tf [(18.)] TJ ET BT 43.553 630.333 Td /F1 9.8 Tf [(Cepeda C, Ariano MA, Calvert CR, Flores-Hernndez J, Chandler SH, Leavitt BR, Hayden MR, Levine MS. NMDA receptor )] TJ ET BT 26.250 618.429 Td /F1 9.8 Tf [(function in mouse models of Huntington disease. J Neurosci Res. 2001 Nov 15;66\(4\):525-39. PubMed PMID:11746372.)] TJ ET BT 26.250 599.024 Td /F1 9.8 Tf [(19.)] TJ ET BT 43.553 599.024 Td /F1 9.8 Tf [(Cepeda C, Hurst RS, Calvert CR, Hernndez-Echeagaray E, Nguyen OK, Jocoy E, Christian LJ, Ariano MA, Levine MS. )] TJ ET BT 26.250 587.119 Td /F1 9.8 Tf [(Transient and progressive electrophysiological alterations in the corticostriatal pathway in a mouse model of Huntington's )] TJ ET BT 26.250 575.214 Td /F1 9.8 Tf [(disease. J Neurosci. 2003 Feb 1;23\(3\):961-9. PubMed PMID:12574425.)] TJ ET BT 26.250 555.810 Td /F1 9.8 Tf [(20.)] TJ ET BT 43.553 555.810 Td /F1 9.8 Tf [(Abada YS, Schreiber R, Ellenbroek B. Motor, emotional and cognitive deficits in adult BACHD mice: a model for )] TJ ET BT 26.250 543.905 Td /F1 9.8 Tf [(Huntington's disease. Behav Brain Res. 2013 Feb 1;238:243-51. PubMed PMID:23123142.)] TJ ET BT 26.250 524.500 Td /F1 9.8 Tf [(21.)] TJ ET BT 43.553 524.500 Td /F1 9.8 Tf [(Kudo T, Schroeder A, Loh DH, Kuljis D, Jordan MC, Roos KP, Colwell CS. Dysfunctions in circadian behavior and )] TJ ET BT 26.250 512.595 Td /F1 9.8 Tf [(physiology in mouse models of Huntington's disease. Exp Neurol. 2011 Mar;228\(1\):80-90. PubMed PMID:21184755.)] TJ ET BT 26.250 493.191 Td /F1 9.8 Tf [(22.)] TJ ET BT 43.553 493.191 Td /F1 9.8 Tf [(Morton AJ, Lagan MA, Skepper JN, Dunnett SB. Progressive formation of inclusions in the striatum and hippocampus of )] TJ ET BT 26.250 481.286 Td /F1 9.8 Tf [(mice transgenic for the human Huntington's disease mutation. J Neurocytol. 2000 Sep;29\(9\):679-702. PubMed PMID:11353291.)] TJ ET BT 26.250 461.881 Td /F1 9.8 Tf [(23.)] TJ ET BT 43.553 461.881 Td /F1 9.8 Tf [(NRC \(1996\) Guide for the care and use of laboratory animals. National Academy, Washington, DC.)] TJ ET BT 26.250 442.476 Td /F1 9.8 Tf [(24.)] TJ ET BT 43.553 442.476 Td /F1 9.8 Tf [(Jacobs GH, Fenwick JC, Calderone JB, Deeb SS. Human cone pigment expressed in transgenic mice yields altered vision. )] TJ ET BT 26.250 430.572 Td /F1 9.8 Tf [(J Neurosci. 1999 Apr 15;19\(8\):3258-65. PubMed PMID:10191338.)] TJ ET BT 26.250 411.167 Td /F1 9.8 Tf [(25.)] TJ ET BT 43.553 411.167 Td /F1 9.8 Tf [(Rudenko O, Tkach V, Berezin V, Bock E. Detection of early behavioral markers of Huntington's disease in R6/2 mice )] TJ ET BT 26.250 399.262 Td /F1 9.8 Tf [(employing an automated social home cage. Behav Brain Res. 2009 Nov 5;203\(2\):188-99. PubMed PMID:19410605.)] TJ ET BT 26.250 379.857 Td /F1 9.8 Tf [(26.)] TJ ET BT 43.553 379.857 Td /F1 9.8 Tf [(Bckman L, Robins-Wahlin TB, Lundin A, Ginovart N, Farde L. Cognitive deficits in Huntington's disease are predicted by )] TJ ET BT 26.250 367.953 Td /F1 9.8 Tf [(dopaminergic PET markers and brain volumes. Brain. 1997 Dec;120 \( Pt 12\):2207-17. PubMed PMID:9448576.)] TJ ET BT 26.250 348.548 Td /F1 9.8 Tf [(27.)] TJ ET BT 43.553 348.548 Td /F1 9.8 Tf [(Lange KW, Sahakian BJ, Quinn NP, Marsden CD, Robbins TW. Comparison of executive and visuospatial memory function )] TJ ET BT 26.250 336.643 Td /F1 9.8 Tf [(in Huntington's disease and dementia of Alzheimer type matched for degree of dementia. J Neurol Neurosurg Psychiatry. 1995 )] TJ ET BT 26.250 324.738 Td /F1 9.8 Tf [(May;58\(5\):598-606. PubMed PMID:7745410.)] TJ ET BT 26.250 305.334 Td /F1 9.8 Tf [(28.)] TJ ET BT 43.553 305.334 Td /F1 9.8 Tf [(Lawrence AD, Sahakian BJ, Rogers RD, Hodge JR, Robbins TW. Discrimination, reversal, and shift learning in Huntington's )] TJ ET BT 26.250 293.429 Td /F1 9.8 Tf [(disease: mechanisms of impaired response selection. Neuropsychologia. 1999 Nov;37\(12\):1359-74. PubMed PMID:10606011.)] TJ ET BT 26.250 274.024 Td /F1 9.8 Tf [(29.)] TJ ET BT 43.553 274.024 Td /F1 9.8 Tf [(Oscar-Berman M, Zola-Morgan SM, Oberg RG, Bonner RT. Comparative neuropsychology and Korsakoff's syndrome. III--)] TJ ET BT 26.250 262.119 Td /F1 9.8 Tf [(Delayed response, delayed alternation and DRL performance. Neuropsychologia. 1982;20\(2\):187-202. PubMed PMID:6211634.)] TJ ET BT 26.250 242.715 Td /F1 9.8 Tf [(30.)] TJ ET BT 43.553 242.715 Td /F1 9.8 Tf [(Oakeshott S, Port RG, Cummins-Sutphen J, Watson-Johnson J, Ramboz S, Park L, Howland D, Brunner D. HD mouse )] TJ ET BT 26.250 230.810 Td /F1 9.8 Tf [(models reveal clear deficits in learning to perform a simple instrumental response. PLoS Curr. 2011 Nov 30;3:RRN1282. )] TJ ET BT 26.250 218.905 Td /F1 9.8 Tf [(PubMed PMID:22512000.)] TJ ET BT 26.250 199.500 Td /F1 9.8 Tf [(31.)] TJ ET BT 43.553 199.500 Td /F1 9.8 Tf [(Oakeshott S, Port R, Cummins-Sutphen J, Berger J, Watson-Johnson J, Ramboz S, Paterson N, Kwak S, Howland D, )] TJ ET BT 26.250 187.596 Td /F1 9.8 Tf [(Brunner D. A mixed fixed ratio/progressive ratio procedure reveals an apathy phenotype in the BAC HD and the z_Q175 KI )] TJ ET BT 26.250 175.691 Td /F1 9.8 Tf [(mouse models of Huntingtons disease. PLOS Currents Huntington Disease. 2012 Apr 25 [last modified: 2012 Sep 4]. Edition 1. )] TJ ET BT 26.250 163.786 Td /F1 9.8 Tf [(doi: 10.1371/4f972cffe82c0.)] TJ ET BT 26.250 144.381 Td /F1 9.8 Tf [(32.)] TJ ET BT 43.553 144.381 Td /F1 9.8 Tf [(Wood NI, Goodman AO, van der Burg JM, Gazeau V, Brundin P, Bjrkqvist M, Petersn A, Tabrizi SJ, Barker RA, Morton )] TJ ET BT 26.250 132.477 Td /F1 9.8 Tf [(AJ. Increased thirst and drinking in Huntington's disease and the R6/2 mouse. Brain Res Bull. 2008 May 15;76\(1-2\):70-9. )] TJ ET BT 26.250 120.572 Td /F1 9.8 Tf [(PubMed PMID:18395613.)] TJ ET BT 26.250 101.167 Td /F1 9.8 Tf [(33.)] TJ ET BT 43.553 101.167 Td /F1 9.8 Tf [(Gray M, Shirasaki DI, Cepeda C, Andr VM, Wilburn B, Lu XH, Tao J, Yamazaki I, Li SH, Sun YE, Li XJ, Levine MS, Yang )] TJ ET BT 26.250 89.262 Td /F1 9.8 Tf [(XW. Full-length human mutant huntingtin with a stable polyglutamine repeat can elicit progressive and selective )] TJ ET BT 26.250 77.358 Td /F1 9.8 Tf [(neuropathogenesis in BACHD mice. J Neurosci. 2008 Jun 11;28\(24\):6182-95. PubMed PMID:18550760.)] TJ ET BT 26.250 57.953 Td /F1 9.8 Tf [(34.)] TJ ET BT 43.553 57.953 Td /F1 9.8 Tf [(Hostetter, R. \(1966\). Time of day effects on learning and open field activity. Psychon. Sci., 5, 257-258.)] TJ ET Q q 15.000 48.072 577.500 728.928 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(PubMed PMID:22512000.)] TJ ET BT 26.250 748.071 Td /F1 9.8 Tf [(15.)] TJ ET BT 43.553 748.071 Td /F1 9.8 Tf [(Balci F, Day M, Rooney A, Brunner D. Disrupted temporal control in the R6/2 mouse model of Huntington's disease. Behav )] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(Neurosci. 2009 Dec;123\(6\):1353-8. PubMed PMID:20001119.)] TJ ET BT 26.250 716.762 Td /F1 9.8 Tf [(16.)] TJ ET BT 43.553 716.762 Td /F1 9.8 Tf [(Grote HE, Bull ND, Howard ML, van Dellen A, Blakemore C, Bartlett PF, Hannan AJ. Cognitive disorders and neurogenesis )] TJ ET BT 26.250 704.857 Td /F1 9.8 Tf [(deficits in Huntington's disease mice are rescued by fluoxetine. Eur J Neurosci. 2005 Oct;22\(8\):2081-8. PubMed )] TJ ET BT 26.250 692.952 Td /F1 9.8 Tf [(PMID:16262645.)] TJ ET BT 26.250 673.548 Td /F1 9.8 Tf [(17.)] TJ ET BT 43.553 673.548 Td /F1 9.8 Tf [(Murphy KP, Carter RJ, Lione LA, Mangiarini L, Mahal A, Bates GP, Dunnett SB, Morton AJ. Abnormal synaptic plasticity )] TJ ET BT 26.250 661.643 Td /F1 9.8 Tf [(and impaired spatial cognition in mice transgenic for exon 1 of the human Huntington's disease mutation. J Neurosci. 2000 Jul )] TJ ET BT 26.250 649.738 Td /F1 9.8 Tf [(1;20\(13\):5115-23. PubMed PMID:10864968.)] TJ ET BT 26.250 630.333 Td /F1 9.8 Tf [(18.)] TJ ET BT 43.553 630.333 Td /F1 9.8 Tf [(Cepeda C, Ariano MA, Calvert CR, Flores-Hernndez J, Chandler SH, Leavitt BR, Hayden MR, Levine MS. NMDA receptor )] TJ ET BT 26.250 618.429 Td /F1 9.8 Tf [(function in mouse models of Huntington disease. J Neurosci Res. 2001 Nov 15;66\(4\):525-39. PubMed PMID:11746372.)] TJ ET BT 26.250 599.024 Td /F1 9.8 Tf [(19.)] TJ ET BT 43.553 599.024 Td /F1 9.8 Tf [(Cepeda C, Hurst RS, Calvert CR, Hernndez-Echeagaray E, Nguyen OK, Jocoy E, Christian LJ, Ariano MA, Levine MS. )] TJ ET BT 26.250 587.119 Td /F1 9.8 Tf [(Transient and progressive electrophysiological alterations in the corticostriatal pathway in a mouse model of Huntington's )] TJ ET BT 26.250 575.214 Td /F1 9.8 Tf [(disease. J Neurosci. 2003 Feb 1;23\(3\):961-9. PubMed PMID:12574425.)] TJ ET BT 26.250 555.810 Td /F1 9.8 Tf [(20.)] TJ ET BT 43.553 555.810 Td /F1 9.8 Tf [(Abada YS, Schreiber R, Ellenbroek B. Motor, emotional and cognitive deficits in adult BACHD mice: a model for )] TJ ET BT 26.250 543.905 Td /F1 9.8 Tf [(Huntington's disease. Behav Brain Res. 2013 Feb 1;238:243-51. PubMed PMID:23123142.)] TJ ET BT 26.250 524.500 Td /F1 9.8 Tf [(21.)] TJ ET BT 43.553 524.500 Td /F1 9.8 Tf [(Kudo T, Schroeder A, Loh DH, Kuljis D, Jordan MC, Roos KP, Colwell CS. Dysfunctions in circadian behavior and )] TJ ET BT 26.250 512.595 Td /F1 9.8 Tf [(physiology in mouse models of Huntington's disease. Exp Neurol. 2011 Mar;228\(1\):80-90. PubMed PMID:21184755.)] TJ ET BT 26.250 493.191 Td /F1 9.8 Tf [(22.)] TJ ET BT 43.553 493.191 Td /F1 9.8 Tf [(Morton AJ, Lagan MA, Skepper JN, Dunnett SB. Progressive formation of inclusions in the striatum and hippocampus of )] TJ ET BT 26.250 481.286 Td /F1 9.8 Tf [(mice transgenic for the human Huntington's disease mutation. J Neurocytol. 2000 Sep;29\(9\):679-702. PubMed PMID:11353291.)] TJ ET BT 26.250 461.881 Td /F1 9.8 Tf [(23.)] TJ ET BT 43.553 461.881 Td /F1 9.8 Tf [(NRC \(1996\) Guide for the care and use of laboratory animals. National Academy, Washington, DC.)] TJ ET BT 26.250 442.476 Td /F1 9.8 Tf [(24.)] TJ ET BT 43.553 442.476 Td /F1 9.8 Tf [(Jacobs GH, Fenwick JC, Calderone JB, Deeb SS. Human cone pigment expressed in transgenic mice yields altered vision. )] TJ ET BT 26.250 430.572 Td /F1 9.8 Tf [(J Neurosci. 1999 Apr 15;19\(8\):3258-65. PubMed PMID:10191338.)] TJ ET BT 26.250 411.167 Td /F1 9.8 Tf [(25.)] TJ ET BT 43.553 411.167 Td /F1 9.8 Tf [(Rudenko O, Tkach V, Berezin V, Bock E. Detection of early behavioral markers of Huntington's disease in R6/2 mice )] TJ ET BT 26.250 399.262 Td /F1 9.8 Tf [(employing an automated social home cage. Behav Brain Res. 2009 Nov 5;203\(2\):188-99. PubMed PMID:19410605.)] TJ ET BT 26.250 379.857 Td /F1 9.8 Tf [(26.)] TJ ET BT 43.553 379.857 Td /F1 9.8 Tf [(Bckman L, Robins-Wahlin TB, Lundin A, Ginovart N, Farde L. Cognitive deficits in Huntington's disease are predicted by )] TJ ET BT 26.250 367.953 Td /F1 9.8 Tf [(dopaminergic PET markers and brain volumes. Brain. 1997 Dec;120 \( Pt 12\):2207-17. PubMed PMID:9448576.)] TJ ET BT 26.250 348.548 Td /F1 9.8 Tf [(27.)] TJ ET BT 43.553 348.548 Td /F1 9.8 Tf [(Lange KW, Sahakian BJ, Quinn NP, Marsden CD, Robbins TW. Comparison of executive and visuospatial memory function )] TJ ET BT 26.250 336.643 Td /F1 9.8 Tf [(in Huntington's disease and dementia of Alzheimer type matched for degree of dementia. J Neurol Neurosurg Psychiatry. 1995 )] TJ ET BT 26.250 324.738 Td /F1 9.8 Tf [(May;58\(5\):598-606. PubMed PMID:7745410.)] TJ ET BT 26.250 305.334 Td /F1 9.8 Tf [(28.)] TJ ET BT 43.553 305.334 Td /F1 9.8 Tf [(Lawrence AD, Sahakian BJ, Rogers RD, Hodge JR, Robbins TW. Discrimination, reversal, and shift learning in Huntington's )] TJ ET BT 26.250 293.429 Td /F1 9.8 Tf [(disease: mechanisms of impaired response selection. Neuropsychologia. 1999 Nov;37\(12\):1359-74. PubMed PMID:10606011.)] TJ ET BT 26.250 274.024 Td /F1 9.8 Tf [(29.)] TJ ET BT 43.553 274.024 Td /F1 9.8 Tf [(Oscar-Berman M, Zola-Morgan SM, Oberg RG, Bonner RT. Comparative neuropsychology and Korsakoff's syndrome. III--)] TJ ET BT 26.250 262.119 Td /F1 9.8 Tf [(Delayed response, delayed alternation and DRL performance. Neuropsychologia. 1982;20\(2\):187-202. PubMed PMID:6211634.)] TJ ET BT 26.250 242.715 Td /F1 9.8 Tf [(30.)] TJ ET BT 43.553 242.715 Td /F1 9.8 Tf [(Oakeshott S, Port RG, Cummins-Sutphen J, Watson-Johnson J, Ramboz S, Park L, Howland D, Brunner D. HD mouse )] TJ ET BT 26.250 230.810 Td /F1 9.8 Tf [(models reveal clear deficits in learning to perform a simple instrumental response. PLoS Curr. 2011 Nov 30;3:RRN1282. )] TJ ET BT 26.250 218.905 Td /F1 9.8 Tf [(PubMed PMID:22512000.)] TJ ET BT 26.250 199.500 Td /F1 9.8 Tf [(31.)] TJ ET BT 43.553 199.500 Td /F1 9.8 Tf [(Oakeshott S, Port R, Cummins-Sutphen J, Berger J, Watson-Johnson J, Ramboz S, Paterson N, Kwak S, Howland D, )] TJ ET BT 26.250 187.596 Td /F1 9.8 Tf [(Brunner D. A mixed fixed ratio/progressive ratio procedure reveals an apathy phenotype in the BAC HD and the z_Q175 KI )] TJ ET BT 26.250 175.691 Td /F1 9.8 Tf [(mouse models of Huntingtons disease. PLOS Currents Huntington Disease. 2012 Apr 25 [last modified: 2012 Sep 4]. Edition 1. )] TJ ET BT 26.250 163.786 Td /F1 9.8 Tf [(doi: 10.1371/4f972cffe82c0.)] TJ ET BT 26.250 144.381 Td /F1 9.8 Tf [(32.)] TJ ET BT 43.553 144.381 Td /F1 9.8 Tf [(Wood NI, Goodman AO, van der Burg JM, Gazeau V, Brundin P, Bjrkqvist M, Petersn A, Tabrizi SJ, Barker RA, Morton )] TJ ET BT 26.250 132.477 Td /F1 9.8 Tf [(AJ. Increased thirst and drinking in Huntington's disease and the R6/2 mouse. Brain Res Bull. 2008 May 15;76\(1-2\):70-9. )] TJ ET BT 26.250 120.572 Td /F1 9.8 Tf [(PubMed PMID:18395613.)] TJ ET BT 26.250 101.167 Td /F1 9.8 Tf [(33.)] TJ ET BT 43.553 101.167 Td /F1 9.8 Tf [(Gray M, Shirasaki DI, Cepeda C, Andr VM, Wilburn B, Lu XH, Tao J, Yamazaki I, Li SH, Sun YE, Li XJ, Levine MS, Yang )] TJ ET BT 26.250 89.262 Td /F1 9.8 Tf [(XW. Full-length human mutant huntingtin with a stable polyglutamine repeat can elicit progressive and selective )] TJ ET BT 26.250 77.358 Td /F1 9.8 Tf [(neuropathogenesis in BACHD mice. J Neurosci. 2008 Jun 11;28\(24\):6182-95. PubMed PMID:18550760.)] TJ ET BT 26.250 57.953 Td /F1 9.8 Tf [(34.)] TJ ET BT 43.553 57.953 Td /F1 9.8 Tf [(Hostetter, R. \(1966\). Time of day effects on learning and open field activity. Psychon. Sci., 5, 257-258.)] TJ ET Q q 15.000 48.072 577.500 728.928 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(PubMed PMID:22512000.)] TJ ET BT 26.250 748.071 Td /F1 9.8 Tf [(15.)] TJ ET BT 43.553 748.071 Td /F1 9.8 Tf [(Balci F, Day M, Rooney A, Brunner D. Disrupted temporal control in the R6/2 mouse model of Huntington's disease. Behav )] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(Neurosci. 2009 Dec;123\(6\):1353-8. PubMed PMID:20001119.)] TJ ET BT 26.250 716.762 Td /F1 9.8 Tf [(16.)] TJ ET BT 43.553 716.762 Td /F1 9.8 Tf [(Grote HE, Bull ND, Howard ML, van Dellen A, Blakemore C, Bartlett PF, Hannan AJ. Cognitive disorders and neurogenesis )] TJ ET BT 26.250 704.857 Td /F1 9.8 Tf [(deficits in Huntington's disease mice are rescued by fluoxetine. Eur J Neurosci. 2005 Oct;22\(8\):2081-8. PubMed )] TJ ET BT 26.250 692.952 Td /F1 9.8 Tf [(PMID:16262645.)] TJ ET BT 26.250 673.548 Td /F1 9.8 Tf [(17.)] TJ ET BT 43.553 673.548 Td /F1 9.8 Tf [(Murphy KP, Carter RJ, Lione LA, Mangiarini L, Mahal A, Bates GP, Dunnett SB, Morton AJ. Abnormal synaptic plasticity )] TJ ET BT 26.250 661.643 Td /F1 9.8 Tf [(and impaired spatial cognition in mice transgenic for exon 1 of the human Huntington's disease mutation. J Neurosci. 2000 Jul )] TJ ET BT 26.250 649.738 Td /F1 9.8 Tf [(1;20\(13\):5115-23. PubMed PMID:10864968.)] TJ ET BT 26.250 630.333 Td /F1 9.8 Tf [(18.)] TJ ET BT 43.553 630.333 Td /F1 9.8 Tf [(Cepeda C, Ariano MA, Calvert CR, Flores-Hernndez J, Chandler SH, Leavitt BR, Hayden MR, Levine MS. NMDA receptor )] TJ ET BT 26.250 618.429 Td /F1 9.8 Tf [(function in mouse models of Huntington disease. J Neurosci Res. 2001 Nov 15;66\(4\):525-39. PubMed PMID:11746372.)] TJ ET BT 26.250 599.024 Td /F1 9.8 Tf [(19.)] TJ ET BT 43.553 599.024 Td /F1 9.8 Tf [(Cepeda C, Hurst RS, Calvert CR, Hernndez-Echeagaray E, Nguyen OK, Jocoy E, Christian LJ, Ariano MA, Levine MS. )] TJ ET BT 26.250 587.119 Td /F1 9.8 Tf [(Transient and progressive electrophysiological alterations in the corticostriatal pathway in a mouse model of Huntington's )] TJ ET BT 26.250 575.214 Td /F1 9.8 Tf [(disease. J Neurosci. 2003 Feb 1;23\(3\):961-9. PubMed PMID:12574425.)] TJ ET BT 26.250 555.810 Td /F1 9.8 Tf [(20.)] TJ ET BT 43.553 555.810 Td /F1 9.8 Tf [(Abada YS, Schreiber R, Ellenbroek B. Motor, emotional and cognitive deficits in adult BACHD mice: a model for )] TJ ET BT 26.250 543.905 Td /F1 9.8 Tf [(Huntington's disease. Behav Brain Res. 2013 Feb 1;238:243-51. PubMed PMID:23123142.)] TJ ET BT 26.250 524.500 Td /F1 9.8 Tf [(21.)] TJ ET BT 43.553 524.500 Td /F1 9.8 Tf [(Kudo T, Schroeder A, Loh DH, Kuljis D, Jordan MC, Roos KP, Colwell CS. Dysfunctions in circadian behavior and )] TJ ET BT 26.250 512.595 Td /F1 9.8 Tf [(physiology in mouse models of Huntington's disease. Exp Neurol. 2011 Mar;228\(1\):80-90. PubMed PMID:21184755.)] TJ ET BT 26.250 493.191 Td /F1 9.8 Tf [(22.)] TJ ET BT 43.553 493.191 Td /F1 9.8 Tf [(Morton AJ, Lagan MA, Skepper JN, Dunnett SB. Progressive formation of inclusions in the striatum and hippocampus of )] TJ ET BT 26.250 481.286 Td /F1 9.8 Tf [(mice transgenic for the human Huntington's disease mutation. J Neurocytol. 2000 Sep;29\(9\):679-702. PubMed PMID:11353291.)] TJ ET BT 26.250 461.881 Td /F1 9.8 Tf [(23.)] TJ ET BT 43.553 461.881 Td /F1 9.8 Tf [(NRC \(1996\) Guide for the care and use of laboratory animals. National Academy, Washington, DC.)] TJ ET BT 26.250 442.476 Td /F1 9.8 Tf [(24.)] TJ ET BT 43.553 442.476 Td /F1 9.8 Tf [(Jacobs GH, Fenwick JC, Calderone JB, Deeb SS. Human cone pigment expressed in transgenic mice yields altered vision. )] TJ ET BT 26.250 430.572 Td /F1 9.8 Tf [(J Neurosci. 1999 Apr 15;19\(8\):3258-65. PubMed PMID:10191338.)] TJ ET BT 26.250 411.167 Td /F1 9.8 Tf [(25.)] TJ ET BT 43.553 411.167 Td /F1 9.8 Tf [(Rudenko O, Tkach V, Berezin V, Bock E. Detection of early behavioral markers of Huntington's disease in R6/2 mice )] TJ ET BT 26.250 399.262 Td /F1 9.8 Tf [(employing an automated social home cage. Behav Brain Res. 2009 Nov 5;203\(2\):188-99. PubMed PMID:19410605.)] TJ ET BT 26.250 379.857 Td /F1 9.8 Tf [(26.)] TJ ET BT 43.553 379.857 Td /F1 9.8 Tf [(Bckman L, Robins-Wahlin TB, Lundin A, Ginovart N, Farde L. Cognitive deficits in Huntington's disease are predicted by )] TJ ET BT 26.250 367.953 Td /F1 9.8 Tf [(dopaminergic PET markers and brain volumes. Brain. 1997 Dec;120 \( Pt 12\):2207-17. PubMed PMID:9448576.)] TJ ET BT 26.250 348.548 Td /F1 9.8 Tf [(27.)] TJ ET BT 43.553 348.548 Td /F1 9.8 Tf [(Lange KW, Sahakian BJ, Quinn NP, Marsden CD, Robbins TW. Comparison of executive and visuospatial memory function )] TJ ET BT 26.250 336.643 Td /F1 9.8 Tf [(in Huntington's disease and dementia of Alzheimer type matched for degree of dementia. J Neurol Neurosurg Psychiatry. 1995 )] TJ ET BT 26.250 324.738 Td /F1 9.8 Tf [(May;58\(5\):598-606. PubMed PMID:7745410.)] TJ ET BT 26.250 305.334 Td /F1 9.8 Tf [(28.)] TJ ET BT 43.553 305.334 Td /F1 9.8 Tf [(Lawrence AD, Sahakian BJ, Rogers RD, Hodge JR, Robbins TW. Discrimination, reversal, and shift learning in Huntington's )] TJ ET BT 26.250 293.429 Td /F1 9.8 Tf [(disease: mechanisms of impaired response selection. Neuropsychologia. 1999 Nov;37\(12\):1359-74. PubMed PMID:10606011.)] TJ ET BT 26.250 274.024 Td /F1 9.8 Tf [(29.)] TJ ET BT 43.553 274.024 Td /F1 9.8 Tf [(Oscar-Berman M, Zola-Morgan SM, Oberg RG, Bonner RT. Comparative neuropsychology and Korsakoff's syndrome. III--)] TJ ET BT 26.250 262.119 Td /F1 9.8 Tf [(Delayed response, delayed alternation and DRL performance. Neuropsychologia. 1982;20\(2\):187-202. PubMed PMID:6211634.)] TJ ET BT 26.250 242.715 Td /F1 9.8 Tf [(30.)] TJ ET BT 43.553 242.715 Td /F1 9.8 Tf [(Oakeshott S, Port RG, Cummins-Sutphen J, Watson-Johnson J, Ramboz S, Park L, Howland D, Brunner D. HD mouse )] TJ ET BT 26.250 230.810 Td /F1 9.8 Tf [(models reveal clear deficits in learning to perform a simple instrumental response. PLoS Curr. 2011 Nov 30;3:RRN1282. )] TJ ET BT 26.250 218.905 Td /F1 9.8 Tf [(PubMed PMID:22512000.)] TJ ET BT 26.250 199.500 Td /F1 9.8 Tf [(31.)] TJ ET BT 43.553 199.500 Td /F1 9.8 Tf [(Oakeshott S, Port R, Cummins-Sutphen J, Berger J, Watson-Johnson J, Ramboz S, Paterson N, Kwak S, Howland D, )] TJ ET BT 26.250 187.596 Td /F1 9.8 Tf [(Brunner D. A mixed fixed ratio/progressive ratio procedure reveals an apathy phenotype in the BAC HD and the z_Q175 KI )] TJ ET BT 26.250 175.691 Td /F1 9.8 Tf [(mouse models of Huntingtons disease. PLOS Currents Huntington Disease. 2012 Apr 25 [last modified: 2012 Sep 4]. Edition 1. )] TJ ET BT 26.250 163.786 Td /F1 9.8 Tf [(doi: 10.1371/4f972cffe82c0.)] TJ ET BT 26.250 144.381 Td /F1 9.8 Tf [(32.)] TJ ET BT 43.553 144.381 Td /F1 9.8 Tf [(Wood NI, Goodman AO, van der Burg JM, Gazeau V, Brundin P, Bjrkqvist M, Petersn A, Tabrizi SJ, Barker RA, Morton )] TJ ET BT 26.250 132.477 Td /F1 9.8 Tf [(AJ. Increased thirst and drinking in Huntington's disease and the R6/2 mouse. Brain Res Bull. 2008 May 15;76\(1-2\):70-9. )] TJ ET BT 26.250 120.572 Td /F1 9.8 Tf [(PubMed PMID:18395613.)] TJ ET BT 26.250 101.167 Td /F1 9.8 Tf [(33.)] TJ ET BT 43.553 101.167 Td /F1 9.8 Tf [(Gray M, Shirasaki DI, Cepeda C, Andr VM, Wilburn B, Lu XH, Tao J, Yamazaki I, Li SH, Sun YE, Li XJ, Levine MS, Yang )] TJ ET BT 26.250 89.262 Td /F1 9.8 Tf [(XW. Full-length human mutant huntingtin with a stable polyglutamine repeat can elicit progressive and selective )] TJ ET BT 26.250 77.358 Td /F1 9.8 Tf [(neuropathogenesis in BACHD mice. J Neurosci. 2008 Jun 11;28\(24\):6182-95. PubMed PMID:18550760.)] TJ ET BT 26.250 57.953 Td /F1 9.8 Tf [(34.)] TJ ET BT 43.553 57.953 Td /F1 9.8 Tf [(Hostetter, R. \(1966\). Time of day effects on learning and open field activity. Psychon. Sci., 5, 257-258.)] TJ ET Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(13)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Huntington Disease)] TJ ET Q endstream endobj 587 0 obj << /Type /Page /Parent 3 0 R /Contents 588 0 R >> endobj 588 0 obj << /Length 1739 >> stream 0.271 0.267 0.267 rg q 15.000 718.786 577.500 58.214 re W n 0.271 0.267 0.267 rg BT 26.250 759.976 Td /F1 9.8 Tf [(35.)] TJ ET BT 43.553 759.976 Td /F1 9.8 Tf [(Valentinuzzi VS, Buxton OM, Chang AM, Scarbrough K, Ferrari EA, Takahashi JS, Turek FW. Locomotor response to an )] TJ ET BT 26.250 748.071 Td /F1 9.8 Tf [(open field during C57BL/6J active and inactive phases: differences dependent on conditions of illumination. Physiol Behav. )] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(2000 May;69\(3\):269-75. 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