Name: behavioral assessment of manual dexterity in non-human primates
Uploaded: 2011-11-11T13:00:00
Duration: 16 min
Description: Watch this Scientific Journal Video about Behavioral Assessment of Manual Dexterity in Non-Human Primates at JoVE.com

Авторы хотели бы поблагодарить профессора М. Шваб, д-р П. Фройнд, д-р А. Висс, д-р С. Башир, д-р А. Мир, д-р Дж. Блок, доктор Ф. Брюне, д-р Дж. Aebischer, д-р А. Месарош, д-р В. Goetschman за их вклад в предыдущих экспериментов и анализов. Экспериментальные установки были построены Андре Гайар, Бернард Aebischer и Лоран ДЕНЕГ. В животном объекта, обезьян были помещены под руководством профессиональных опекунов животных: Йозеф Corpataux, Лоран Босси и Жак Майяр. Поведенческие тесты и анализ данных, а также гистология, были проведены с весьма ценный вклад лаборантов: Вероник Море (также веб-мастера), Франсуаза Тингели, Кристин Рулен, Моника Bennefeld, Кристиан Марти и Жоржетта Фишер. Работа выполнена при поддержке Швейцарского национального научного фонда, гранты Нет 31-61857.00, 310000-110005, 132465-31003A (ЭМИ), 310030-118357, 104061-31003A (TW), 310030-120411 (ABS), PZ00P3_121646 (ES), Novartis Foundation; Национальный центр компетенции в области научных исследований (НКРС) на тему &quot;Нейронные пластичности и ремонт&quot;.

<ol>
	<li>Lemon, R. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Descending+pathways+in+motor+control.">Descending pathways in motor control.</a> Annu. Rev. Neurosci. 31, 195-218 (2008).</li><li>Courtine, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Can+experiments+in+nonhuman+primates+expedite+the+translation+of+treatments+for+spinal+cord+injury+in+humans?.">Can experiments in nonhuman primates expedite the translation of treatments for spinal cord injury in humans?.</a> Nat. Med. 13, 561-566 (2007).</li><li>Capitanio, J. P., Emborg, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contributions+of+non-human+primates+to+neuroscience+research.">Contributions of non-human primates to neuroscience research.</a> Lancet. 371, 1126-1135 (2008).</li><li>Brinkman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Supplementary+motor+area+of+the+monkey's+cerebral+cortex:+short-+and+long-term+deficits+after+unilateral+ablation+and+the+effects+of+subsequent+callosal+section.">Supplementary motor area of the monkey's cerebral cortex: short- and long-term deficits after unilateral ablation and the effects of subsequent callosal section.</a> J. Neurosci. 4, 918-929 (1984).</li><li>Brinkman, J., Kuypers, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebral+control+of+contralateral+and+ipsilateral+arm,+hand+and+finger+movements+in+the+split-brain+rhesus+monkey.">Cerebral control of contralateral and ipsilateral arm, hand and finger movements in the split-brain rhesus monkey.</a> Brain. , 653-674 (1973).</li><li>Kazennikov, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporal+structure+of+a+bimanual+goal-directed+movement+sequence+in+monkeys.">Temporal structure of a bimanual goal-directed movement sequence in monkeys.</a> Eur. J. Neurosci. 6, 203-210 (1994).</li><li>Kazennikov, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+activity+of+supplementary+and+primary+motor+areas+in+monkeys+and+its+relation+to+bimanual+and+unimanual+movement+sequences.">Neural activity of supplementary and primary motor areas in monkeys and its relation to bimanual and unimanual movement sequences.</a> Neuroscience. 89, 661-674 (1999).</li><li>Kermadi, I., Liu, Y., Tempini, A., Rouiller, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+reversible+inactivation+of+the+supplementary+motor+area+(SMA)+on+unimanual+grasp+and+bimanual+pull+and+grasp+performance+in+monkeys.">Effects of reversible inactivation of the supplementary motor area (SMA) on unimanual grasp and bimanual pull and grasp performance in monkeys.</a> Somatosens. Mot. Res. 14, 268-280 (1997).</li><li>Kermadi, I., Liu, Y., Tempini, A., Calciati, E., Rouiller, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+activity+in+the+primate+supplementary+motor+area+and+the+primary+motor+cortex+in+relation+to+spatio-temporal+bimanual+coordination.">Neuronal activity in the primate supplementary motor area and the primary motor cortex in relation to spatio-temporal bimanual coordination.</a> Somatosens. Mot. Res. 15, 287-308 (1998).</li><li>Kermadi, I., Liu, Y., Rouiller, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Do+bimanual+motor+actions+involve+the+dorsal+premotor+(PMd),+cingulate+(CMA)+and+posterior+parietal+(PPC)+cortices?+Comparison+with+primary+and+supplementary+motor+cortical+areas.">Do bimanual motor actions involve the dorsal premotor (PMd), cingulate (CMA) and posterior parietal (PPC) cortices? Comparison with primary and supplementary motor cortical areas.</a> Somatosensory and Motor Research. 17, 255-271 (2000).</li><li>Wannier, T., Liu, J., Morel, A., Jouffrais, C., Rouiller, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+activity+in+primate+striatum+and+pallidum+related+to+bimanual+motor+actions.">Neuronal activity in primate striatum and pallidum related to bimanual motor actions.</a> NeuroReport. 13, 143-147 (2002).</li><li>Rouiller, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dexterity+in+adult+monkeys+following+early+lesion+of+the+motor+cortical+hand+area:+the+role+of+cortex+adjacent+to+the+lesion.">Dexterity in adult monkeys following early lesion of the motor cortical hand area: the role of cortex adjacent to the lesion.</a> Eur. J. Neurosci. 10, 729-740 (1998).</li><li>Liu, Y., Rouiller, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+recovery+of+dexterity+following+unilateral+lesion+of+the+sensorimotor+cortex+in+adult+monkeys.">Mechanisms of recovery of dexterity following unilateral lesion of the sensorimotor cortex in adult monkeys.</a> Exp. Brain. Res. 128, 149-159 (1999).</li><li>Freund, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nogo-A-specific+antibody+treatment+enhances+sprouting+and+functional+recovery+after+cervical+lesion+in+adult+primates.">Nogo-A-specific antibody treatment enhances sprouting and functional recovery after cervical lesion in adult primates.</a> Nature. Med. 12, 790-792 (2006).</li><li>Freund, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anti-Nogo-A+antibody+treatment+promotes+recovery+of+manual+dexterity+after+unilateral+cervical+lesion+in+adult+primates--re-examination+and+extension+of+behavioral+data.">Anti-Nogo-A antibody treatment promotes recovery of manual dexterity after unilateral cervical lesion in adult primates--re-examination and extension of behavioral data.</a> Eur. J. Neurosci. 29, 983-996 (2009).</li><li>Kaeser, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+Unilateral+Motor+Cortex+Lesion+on+Ipsilesional+Hand's+Reach+and+Grasp+Performance+in+Monkeys:+Relationship+With+Recovery+in+the+Contralesional+Hand.">Effects of Unilateral Motor Cortex Lesion on Ipsilesional Hand's Reach and Grasp Performance in Monkeys: Relationship With Recovery in the Contralesional Hand.</a> J. Neurophysiol. 103, 1603-1645 (2010).</li><li>Kaeser, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autologous+adult+cortical+cell+transplantation+enhances+functional+recovery+following+unilateral+lesion+of+motor+cortex+in+primates:+a+pilot+study.">Autologous adult cortical cell transplantation enhances functional recovery following unilateral lesion of motor cortex in primates: a pilot study.</a> Neurosurgery. 68, 1405-1417 (2011).</li><li>Bashir, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short-term+effects+of+unilateral+lesion+of+the+primary+motor+cortex+(M1)+on+Ipsilesional+hand+dexterity+in+adult+macaque+monkeys.">Short-term effects of unilateral lesion of the primary motor cortex (M1) on Ipsilesional hand dexterity in adult macaque monkeys.</a> Brain Structure and Function. , (2011).</li><li>Hamadjida, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+anti-Nogo-A+treatment+on+the+reorganization+of+callosal+connectivity+of+the+premotor+cortical+areas+following+unilateral+lesion+of+primary+motor+cortex+(M1)+in+adult+macaque+monkeys.">Influence of anti-Nogo-A treatment on the reorganization of callosal connectivity of the premotor cortical areas following unilateral lesion of primary motor cortex (M1) in adult macaque monkeys.</a> , (2011).</li><li>Jenny, A. B., Inukai, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+of+motor+organization+of+the+monkey+cervical+spinal+cord.">Principles of motor organization of the monkey cervical spinal cord.</a> J. Neurosci. 3, 567-575 (1983).</li><li>Schmidlin, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progressive+plastic+changes+in+the+hand+representation+of+the+primary+motor+cortex+parallel+incomplete+recovery+from+a+unilateral+section+of+the+corticospinal+tract+at+cervical+level+in+monkeys.">Progressive plastic changes in the hand representation of the primary motor cortex parallel incomplete recovery from a unilateral section of the corticospinal tract at cervical level in monkeys.</a> Brain Research. 1017, 172-183 (2004).</li><li>Schmidlin, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+the+hand+representation+in+the+ipsilateral+primary+motor+cortex+following+unilateral+section+of+the+corticospinal+tract+at+cervical+level+in+monkeys.">Reduction of the hand representation in the ipsilateral primary motor cortex following unilateral section of the corticospinal tract at cervical level in monkeys.</a> BMC Neuroscience. 6, 56-56 (2005).</li><li>Prescott, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Refinement+of+the+use+of+food+and+fluid+control+as+motivational+tools+for+macaques+used+in+behavioural+neuroscience+research:+report+of+a+Working+Group+of+the+NC3Rs.">Refinement of the use of food and fluid control as motivational tools for macaques used in behavioural neuroscience research: report of a Working Group of the NC3Rs.</a> J. Neurosci. Methods. 193, 167-188 (2010).</li><li>Darling, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volumetric+effects+of+motor+cortex+injury+on+recovery+of+dexterous+movements.">Volumetric effects of motor cortex injury on recovery of dexterous movements.</a> Exp. Neurol. 220, 90-108 (2009).</li><li>Darling, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimal+forced+use+without+constraint+stimulates+spontaneous+use+of+the+impaired+upper+extremity+following+motor+cortex+injury.">Minimal forced use without constraint stimulates spontaneous use of the impaired upper extremity following motor cortex injury.</a> Exp. Brain. Res. 202, 529-542 (2010).</li><li>McNeal, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+long-term+reorganization+of+the+corticospinal+projection+from+the+supplementary+motor+cortex+following+recovery+from+lateral+motor+cortex+injury.">Selective long-term reorganization of the corticospinal projection from the supplementary motor cortex following recovery from lateral motor cortex injury.</a> J. Comp. Neurol. 518, 586-621 (2010).</li><li>Nishimura, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time-dependent+central+compensatory+mechanisms+of+finger+dexterity+after+spinal+cord+injury.">Time-dependent central compensatory mechanisms of finger dexterity after spinal cord injury.</a> Science. 318, 1150-1155 (2007).</li><li>Pizzimenti, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+reaching+kinematics+and+prehensile+dexterity+in+nonhuman+primates.">Measurement of reaching kinematics and prehensile dexterity in nonhuman primates.</a> J. Neurophysiol. 98, 1015-1029 (2007).</li></ol>

Нет конфликта интересов объявлены.

Хотя настоящее поведенческие задачи были рассмотрены до сих пор в нашей лаборатории в рамках исследований, связанных с поражением шейного шнура или поражения моторной коры с целью протестировать различные процедуры (см. 14,15,17 и <a href="http://www.unifr.ch/neuro/rouiller">http:/ / www.unifr.ch / нейро / rouiller</a> &gt; выберите &quot;исследования&quot; в верхней строке меню, а затем&gt; &quot;двигательной системы&quot;&gt; &quot;восстановление после поражения&quot;), они также могут иметь более широкое применение, как ловкость рук также аспект рассмотреть в других патологий, таких как болезнь Паркинсона (МРТР обезьян) или в случае сенсорных де-афферентации, влияющих проприоцепции и / или осязание (особенно тест 2 в отсутствии визуального контроля). Поведенческие тесты предлагаемые здесь пригодны для исследования управления двигателем дистальных передней лапы, так как участвует в ловкости рук. Специфичность тестов демонстрируютОтсутствие дефицита (за исключением нескольких дней) в случае поражения, которое не нарушить соответствующие компоненты системы управления: действительно, в случае поражения размещены более чем на хвостовом мотонейронов управления мышцы рук, не было дефицита. Уместность теста 1 можно оценить путем сравнения на видео последовательностей, принятые на двух временных точках после поражения кривой восстановления, которые отличаются повышение функционального восстановления на 25%, возможно, в отношении лечения клеточной терапии 17. Несмотря на некоторые первоначальные, относительно короткий этап подготовки в начале (продолжительностью обычно 2-3 месяца), поведенческие тесты предлагаемые здесь относительно &quot;естественного&quot; и прямой, по сравнению с комплексом (например, условный) задачи, для которых подготовка обезьяны может занять около года или больше. Положительное подкрепление основано на твердую пищу, которая менее чувствительна по этической точки зрения, чем вода лишения, как правило, используются в более сложныхсложных задач 23. Существует не нужно лишать обезьян от продуктов питания до получения стабильных и устойчивых результатов. Гранулы, полученные в ходе задачи представляют собой первый доступ к пищи в день поведенческих сессии (предполагается, что обезьяны не ест в течение предыдущего ночь, однако дополнительное питание может быть дано до конца второй половине дня предыдущего дня) . Очень важно, чтобы каждая обезьяна выполняет поведенческие сессии, в то же время суток, а также уважения же последовательно между различными обезьян формирования группы в жилищной комнате. Как обезьяны чувствительны к внешним тревожные события, поведенческие задачи должно проводиться в присутствии фоновой музыки, маскируя потенциальных тревожные шума из соседних помещений или лабораторий. Очень важно, чтобы в течение всего периода эксперимента (от начального обучения до последнего ежедневного экспериментальной сессии, в течение нескольких месяцев, если не годы), данный обезьянаразмещены ежедневно под руководством одного и того же экспериментатора. Настоящий поведенческие тесты, используемые уже несколько лет в нашей лаборатории количественно ловкости рук, в некоторой степени сравнимы с другими испытания ловкости рук недавно сообщалось в литературе 24-28. Существует, однако, крайне необходимо для стандартизации тестов в разных лабораториях (для лучшего сравнения), который является предварительным Цель настоящего доклада. По требованию подробную свойства установок показано здесь для тестов 1-4 могут быть предоставлены соответствующие автора, для того, чтобы повторить их. Помимо вопроса регенеративной медицины (восстановление после поражения спинного мозга или головного мозга), настоящее палитру тестов могут быть пригодны для решения в нормальных, не человеческих приматов проблем развития (например, время хода двигателя развития ловкими движениями) в целях изучения латерализации аспекты (стороны предпочтения / господство) и расшифровать эволюционные вопросы сomparing двигательных способностей различных видов приматов, включая человека в качестве субъекта. Заметим, однако, что размеры аппаратов должны быть адаптированы в соответствии с цифрами &quot;размер (толщина и длина) видов приматов, так как это может повлиять на выполнение задания. В настоящем исследовании, тесты проводились на Macaca fascicularis обезьян, от 2,5 до 8 лет и весом от 2,5 до 8 кг. Длина указательного пальца (используется в первую очередь для манипулирования гранулы) составляет от 32 до 35 мм, в то время как окружность дистальной фаланги (совет) в указательный палец был между 22 и 25 мм в обезьян, включенных в наших исследованиях. Как испытания в предыдущих экспериментах, схватив же тесты предназначены для Macaca mulatta также. Все эксперименты проводились в соответствии с Руководство по уходу и использованию лабораторных животных (1996) и утверждаются местными (Швейцария) ветеринарными органами. Все экспериментальные процедуры обезьяныс, а также условия содержания в животных объекта, были подробно описаны в последних докладах из нашей лаборатории: см. ссылки 12-18.

behavioral assessment of manual dexterity in non-human primates

Кортикоспинальных (CS) тракта анатомической поддержкой изысканные способность двигателя умело манипулировать мелкими предметами, прерогатива главным образом приматов 1. В случае поражения, влияющих системы CS проекции на ее происхождение (поражения моторных зон коры) или по своей траектории (шейный поражения мозга), есть драматические потери ловкости рук (руки паралич), как показано в некоторых тетраплегией или гемиплегической пациентов . Хотя есть некоторое спонтанное восстановление функций после такого поражения, она остается весьма ограниченным во взрослом. Различные терапевтические стратегии в настоящее время предлагается (например, клеточной терапии, нейтрализация тормозных аксонального молекул роста, применения факторов роста и др.), которые в основном разработана в грызунов. Однако, прежде чем клиническое применение, часто рекомендуемый для проверки осуществимости, эффективности и безопасности лечения в не человеческих приматов. Это особенно верно, когда целью является восстановление ловкости рук послепоражение центральной нервной системы, а организация двигательной системы грызунов отличается от приматов 1,2. Макак показаны здесь как подходящая модель поведения количественно ловкости рук у приматов, чтобы отразить дефицит в результате поражения двигательной коры или шейный шнур например, измерить степень спонтанного восстановления функций и, когда лечение применяется, оценить сколько это может улучшить функциональное восстановление. Поведенческие оценки ловкости рук основана на четырех различных, дополняют друг друга, достичь понимания и ручных задач (применение высокоточного захвата понять гранул), что требует первоначального обучения взрослых макак. Подготовка животных показали, а также позиционирование по отношению к поведенческие установки. Выполнение типичных обезьяны иллюстрируется для каждой задачи. Сбор и анализ соответствующих показателей, отражающих точные манипуляции стороныТион, а также контроль силы, объясняется и продемонстрировал с представителем результаты. Эти данные размещаются затем в более широком контексте, показывая, как поведенческие данные могут быть использованы для изучения последствий поражения спинного мозга или поражения моторной коры и в какой степени лечение может увеличить спонтанного восстановления функций, сравнивая разных групп обезьян (лечение против мнимого лечения, например). Преимущества и недостатки поведенческих тестов обсуждаются. Настоящий поведенческий подход в соответствии с предыдущими докладами подчеркивая актуальность без человеческих приматов модель в контексте заболевания нервной системы 2,3.

Watch this Scientific Journal Video about Behavioral Assessment of Manual Dexterity in Non-Human Primates at JoVE.com

Behavioral Assessment of Manual Dexterity in Non-Human Primates

Как ловкости рук является прерогативой главным образом приматов, поведенческие задачи были разработаны в макак. Четыре достичь и понять схватыванию задач, измерительные стороны способностью манипулирования и силу, позволяют установить функциональное восстановление после поражения центральной нервной системы и проверить эффект лечения.

Поведенческая оценка ловкость рук в нечеловеческих приматов

The corticospinal (CS) tract is the anatomical support of the exquisite motor ability to skillfully manipulate small objects, a prerogative mainly of ...

behavioral-assessment-of-manual-dexterity-in-non-human-primates

Research

JoVE Journal

Neuroscience

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

Общий метод оценки Инкубация Сахароза Тяга у крыс

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, including rats, hamsters, and mice, are active during the night and relatively inactive during the day. Many other behavioral and physiological measures also exhibit daily rhythms, but in rodents, running-wheel activity serves as a particularly reliable and convenient measure of the output of the master circadian clock, the suprachiasmatic nucleus (SCN) of the hypothalamus. In general, through a process called entrainment, the daily pattern of running-wheel activity will naturally align with the environmental light-dark cycle (LD cycle; e.g. 12 hr-light:12 hr-dark). However circadian rhythms are endogenously generated patterns in behavior that exhibit a ~24 hr period, and persist in constant darkness. Thus, in the absence of an LD cycle, the recording and analysis of running-wheel activity can be used to determine the subjective time-of-day. Because these rhythms are directed by the circadian clock the subjective time-of-day is referred to as the circadian time (CT). In contrast, when an LD cycle is present, the time-of-day that is determined by the environmental LD cycle is called the zeitgeber time (ZT).
Although circadian rhythms in running-wheel activity are typically linked to the SCN clock6-8, circadian oscillators in many other regions of the brain and body9-14 could also be involved in the regulation of daily activity rhythms. For instance, daily rhythms in food-anticipatory activity do not require the SCN15,16 and instead, are correlated with changes in the activity of extra-SCN oscillators17-20. Thus, running-wheel activity recordings can provide important behavioral information not only about the output of the master SCN clock, but also on the activity of extra-SCN oscillators. Below we describe the equipment and methods used to record, analyze and display circadian locomotor activity rhythms in laboratory rodents.

Circadian rhythms in voluntary wheel-running activity in mammals are tightly coupled to the molecular oscillations of a master clock in the brain. As such, these daily rhythms in behavior can be used to study the influence of genetic, pharmacological, and environmental factors on the functioning of this circadian clock.

Запись и анализ циркадных ритмов в рабочем колеса деятельность в Грызуны

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

Assessment of Dendritic Arborization in the Dentate Gyrus of the Hippocampal Region in Mice

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity and extent of dendritic arborization correlates well with the synaptic plasticity in these cells. A reliable method for assessment of dendritic arborization is needed to characterize the deleterious effects of neurological disorders on these structures and to determine the effects of therapeutic interventions. However, quantification of these structures has proven to be a formidable task given their complex and dynamic nature. Fortunately, sophisticated imaging techniques can be paired with conventional staining methods to assess the state of dendritic arborization, providing a more reliable and expeditious means of assessment. Below is an example of how these imaging techniques were paired with staining methods to characterize the dendritic arborization in wild type mice. These complementary imaging methods can be used to qualitatively and quantitatively assess dendritic arborization that span a rather wide area within the hippocampal region.

We describe two methods for visualization and quantification of dendritic arborization in the hippocampus of mouse models: real-time and extended depth of field imaging. While the former method allows sophisticated topographical tracing and quantification of the extent of branching, the latter allows speedy visualization of the dendritic tree.

Оценка ветвления дендритов в зубчатой ​​извилине гиппокампа области у мышей

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity ...

Quantification of Filamentous Actin (F-actin) Puncta in Rat Cortical Neurons

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich structures suggest alterations in synaptic integrity and connectivity. Here we provide a detailed protocol for culturing primary rat cortical neurons, Phalloidin staining for F-actin puncta, and subsequent quantification techniques. First, the frontal cortex of E18 rat embryos are dissociated into low-density cell culture, then the neurons grown in vitro for at least 12-14 days. Following experimental treatment, the cortical neurons are stained with AlexaFluor 488 Phalloidin (to label the dendritic F-actin puncta) and microtubule-associated protein 2 (MAP2; to validate the neuronal cells and dendritic integrity). Finally, specialized software is used to analyze and quantify randomly selected neuronal dendrites. F-actin rich structures are identified on second order dendritic branches (length range 25-75 &#181;m) with continuous MAP2 immunofluorescence. The protocol presented here will be a useful method for investigating changes in dendritic synapse structures subsequent to experimental treatments.

Quantification of Filamentous Actin F-actin Puncta in Rat Cortical Neurons

Filamentous actin (F-actin) plays an important role in spinogenesis, synaptic plasticity, and synaptic stability. Quantification of F-actin puncta is therefore a useful tool to study the integrity of synaptic structures. This protocol describes the procedures of quantifying F-actin puncta labeled with Phalloidin in low-density primary cortical neuronal cultures.

Количественное нитевидных актина (F-актин) Puncta на крысиной корковых нейронов

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich ...

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

In the central nervous system, the D1-alpha subtype receptor (Drd1&#945;) is the most abundant dopamine (DA) receptor, which plays a vital role in regulating neuronal growth and development. However, the mechanisms underlying Drd1&#945; receptor abnormalities mediating behavioral responses and modulating working memory function are still unclear. Using a novel RNA in situ hybridization assay, the current study identified dopamine Drd1&#945; receptor and tyrosine hydroxylase (TH) RNA expression from DA-related circuitry in the nucleus accumbens (NAc) area and substantia nigra region (SNR), respectively. Drd1&#945; expression in the NAc shows a &#34;discrete dot&#34; staining pattern. Clear sex differences in Drd1&#945; expression were observed. In contrast, TH shows a &#34;clustered&#34; staining pattern. Regarding TH expression, female rats displayed a higher signal expression per cell relative to male animals. The methods presented here provide a novel in situ hybridization technique for investigating changes in dopamine system dysfunction during the progression of central nervous system diseases.

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

Identification of dopamine D1-alpha receptor in the nucleus accumbens is critical for clarifying D1 receptor dysfunction during a central nervous system disease. We performed a novel RNA in situ hybridization assay to visualize single RNA molecules in a specific brain area.

Идентификация Дофаминовый рецептор D1-альфа в пределах грызунов прилежащем инновационные РНК в месте обнаружения технологии

In the central nervous system, the D1-alpha subtype receptor (Drd1&#945;) is the most abundant dopamine (DA) receptor, which plays a vital role in ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in neurocognitive disorders. Despite the popularization of prepulse inhibition (PPI) in recent years, many current protocols promote using a percent of control measure, thereby precluding the assessment of temporal processing. The present study used cross-modal PPI and gap prepulse inhibition (gap-PPI) to demonstrate the benefits of employing a range of interstimulus intervals (ISIs) to delineate effects of sensory modality, psychostimulant exposure, and age. Assessment of sensory modality, psychostimulant exposure, and age reveals the utility of an approach varying the interstimulus interval (ISI) to establish the shape of the ISI function, including increases (sharper curve inflections) or decreases (flattening of the response amplitude curve) in startle amplitude. Additionally, shifts in peak response inhibition, suggestive of a differential sensitivity to the manipulation of ISI, are often revealed. Thus, the systematic manipulation of ISI affords a critical opportunity to evaluate temporal processing, which may reveal the underlying neural mechanisms involved in neurocognitive disorders.

Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.

Мощность Interstimulus интервала для оценки временных обработки грызунов

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in ...

Assessment of Long-term Depression Induction in Adult Cerebellar Slices

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from parallel fibers (PF) to Purkinje cells (PC) is considered the basis for motor learning, and deficiencies of both LTD and motor learning are observed in various gene-manipulated animals. Common motor learning sets, such as adaptation of the optokinetic reflex (OKR), the vestibular-ocular reflex (VOR), and rotarod test were used for evaluation of motor learning ability. However, results obtained from the GluA2-carboxy terminus modified knock-in mice demonstrated normal adaptation of the VOR and the OKR, despite lacking PF-LTD. In that report, induction of LTD was only attempted using one type of stimulation protocol at room temperature. Thus, conditions to induce cerebellar LTD were explored in the same knock-in mutants using various protocols at near physiological temperature. Finally, we found stimulation protocols, by which LTD could be induced in these gene-manipulated mice. In this study, a set of protocols are proposed to evaluate LTD-induction, which will more accurately allow examination of the causal relationship between LTD and motor learning. In conclusion, experimental conditions are crucial when evaluating LTD in gene-manipulated mice.

In some gene-manipulated animals, using a single protocol may fail to induce LTD in cerebellar Purkinje cells, and there may be a discrepancy between LTD and motor learning. Multiple protocols are necessary to assess LTD-induction in gene-manipulated animals. Standard protocols are shown.

Оценка долгосрочной депрессии индукции в взрослых церебеллярной ломтики

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from ...

Ballistic Labeling of Pyramidal Neurons in Brain Slices and in Primary Cell Culture

It has been reported that the size and shape of dendritic spines is related to their structural plasticity. To identify the morphological structure of pyramidal neurons and dendritic spines, a ballistic labeling technique can be utilized. In the present protocol, pyramidal neurons are labeled with DilC18(3) dye and analyzed using neuronal reconstruction software to assess neuronal morphology and dendritic spines. To investigate neuronal structure, dendritic branching analysis and Sholl analysis are performed, allowing researchers to draw inferences about dendritic branching complexity and neuronal arbor complexity, respectively. The evaluation of dendritic spines is conducted using an automatic assisted classification algorithm integral to the reconstruction software, which classifies spines into four categories (i.e., thin, mushroom, stubby, filopodia). Furthermore, an additional three parameters (i.e., length, head diameter, and volume) are also chosen to assess alterations in dendritic spine morphology. To validate the potential of wide application of the ballistic labeling technique, pyramidal neurons from in vitro cell culture were successfully labeled. Overall, the ballistic labeling method is unique and useful for visualizing neurons in different brain regions in rats, which in combination with sophisticated reconstruction software, allows researchers to elucidate the possible mechanisms underlying neurocognitive dysfunction.

We present a protocol to label and analyze pyramidal neurons, which is critical for evaluating potential morphological alterations in neurons and dendritic spines that may underlie neurochemical and behavioral abnormalities.

Баллистическая маркировка пирамидальных нейронов в мозговых фрагментах и в первичной культуре клеток

It has been reported that the size and shape of dendritic spines is related to their structural plasticity. To identify the morphological structure of ...

A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered tissue samples. Traditional immunolabeling procedures require cutting the sample into thin sections, which restricts the ability to label and examine intact structures. However, if brain tissue can remain intact during processing, structures and circuits can remain intact for the analysis. Previously established clearing methods take significant time to completely clear the tissue, and the harsh chemicals can often damage sensitive antibodies. The iDISCO method quickly and completely clears tissue, is compatible with many antibodies, and requires no special lab equipment. This technique was initially validated for the use in mice tissue, but the current protocol adapts this method to image hemispheres of control and transgenic rat brains. In addition to this, the present protocol also makes several adjustments to preexisting protocol to provide clearer images with less background staining. Antibodies for Iba-1 and tyrosine hydroxylase were validated in the HIV-1 transgenic rat and in F344/N control rats using the present hydrophobic tissue clearing method. The brain is an interwoven network, where structures work together more often than separately of one another. Analyzing the brain as a whole system as opposed to a combination of individual pieces is the greatest benefit of this whole brain clearing method.

Here we present a hydrophobic tissue clearing method that allows for the viewing of target molecules as part of intact brain structures. This technique has now been validated for F344/N control and HIV-1 transgenic rats of both sexes.

Гидрофобный метод очистки тканей для ткани мозга крыс

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered ...

A Rat Model of EcoHIV Brain Infection

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. Establishment of the EcoHIV infected rat model for studies of drug abuse and neurocognitive disorders, would be beneficial in the study of neuroHIV and HIV-1 associated neurocognitive disorders (HAND). In the present study, we demonstrate the successful creation of a rat model of active HIV infection using chimeric HIV (EcoHIV). First, the lentiviral construct of EcoHIV was packaged in cultured 293 FT cells for 48 hours. Then, the conditional medium was concentrated and titered. Next, we performed bilateral stereotaxic injections of the EcoHIV-EGFP into F344/N rat brain tissue. One week after infection, EGFP fluorescence signals were detected in the infected brain tissue, indicating that EcoHIV successfully induces an active HIV infection in rats. In addition, immunostaining for the microglial cell marker, Iba1, was performed. The results indicated that microglia were the predominant cell type harboring EcoHIV. Furthermore, EcoHIV rats exhibited alterations in temporal processing, a potential underlying neurobehavioral mechanism of HAND as well as synaptic dysfunction eight weeks after infection. Collectively, the present study extends the EcoHIV model of HIV-1 infection to the rat offering a valuable biological system to study HIV-1 viral reservoirs in the brain as well as HAND and associated comorbidities such as drug abuse.

Here, we present a protocol to establish a new rat model of active HIV infection using chimeric HIV (EcoHIV), which is critical for enhancing our understanding of HIV-1 viral reservoirs in the brain and offering a system to study HIV-associated neurocognitive disorders and associated comorbidities (i.e., drug abuse).

Крысиная модель инфекции мозга EcoHIV

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...