我们要感谢新希望研究基金会继续支持和资助我们的调查。我们还要感谢理事会对这项工作的部分财政支持。

本文所述的所有协议都是按照女王大学动物保育委员会的批准进行的。行为测试的频率可以是每周或每月, 或者是两个的组合 (即,每周一次, 当你期望看到最大的差异, 每月在别处)。
注意: 小鼠的行为研究主要是性/年龄/应变依赖性, 因此, 必须收集适当的时间点, 并保持一致的跨实验组。环境因素 (例如,气味、噪音、温度、湿度照明、等) 极大地影响了小鼠的行为和焦虑。尽量减少这些环境因素。
1. 野外试验
注意: 在进行行为测试之前, 动物应该习惯于测试开始前 10-15 分钟的房间。确保行为测试协议遵循适当的个人防护设备和所有当地设施的指导方针。
注: 测试前不进行培训。
<ol><li>确保计算机和经常程序已打开、正在运行, 并正确连接到照相机或开放域框。单击 smartware 程序、打开字段图标和 "确定"。从摄像机选项中选择了 "数字模拟转换器" 源。点击 "检测", 并确保整个开放领域的空间是可视化的, 尺寸是 30 cm x 30 厘米。 注意: 设备布局可能会因空间限制而异, 但要确保计算机和开放的现场设备在光线充足的房间内彼此接近, 以便进行适当的视频捕获。</li>
	<li>单击 "校准"、"快照", 然后 "确定" 以确保在打开的域范围内检测到鼠标。单击 "时间" 以预先设置测试鼠标的时间量;在这种情况下, 设置5分钟。</li>
	<li>通过单击 "主题" 图标输入主题。单击 "计划程序", 并突出显示所有测试对象, 然后单击 "双绿色箭头" 和 "第一阶段, 第一节"; 这将带来所有的主题, 以开始收集数据。</li>
	<li>将鼠标放在打开字段的中心, 然后单击 "开始" 图标以提示程序开始录制。一次只测试一只老鼠。</li>
	<li>一旦5分钟完成, 程序将自动移动到下一个主题。在这种情况下, 请移除已完成测试的鼠标, 并将下一个主题放到打开的字段中。单击 "开始"。</li>
	<li>把设备擦干净对其余的测试对象重复步骤 1.3-1.4。</li>
	<li>为了在所有的实验对象都经过测试后分析数据, 点击屏幕右上方的 "分析" 图标。再次单击 "分析", 然后 "汇总报告"。(可选) 将数据导出到 USB 上。</li>
</ol>2. 网格测试
注: MT 的表面面积和网格大小可能会影响性能。在 MT 上, 如果一只老鼠在第一个十年代的第一个2收集时间点内脱落, 它们将立即被放回网格上进行第二次尝试。这里使用的网格是一个金属丝网, 网孔直径约1.5 毫米。
注意: 建议在一天的同一时间运行这些测试, 因为生理和生物化学参数在一天中波动。体重可能影响这项试验的表现, 因此必须加以考虑。
<ol><li>将鼠标放在网格的中心, 并将网格翻转到至少20厘米以上的透明容器上。一旦反转, 启动计时器。</li>
	<li>时间和记录的滞后时间下降。如果鼠标不属于六十年代, 给它一个分数六十年代和不进行进一步的试验。允许鼠标至少休息5分钟。这是1审判的结束。</li>
	<li>擦拭设备, 并在5分钟内试验间隔, 如果需要, 测试第二个鼠标。重复步骤 2.1-2.2 为试验2和 3, 如果需要 (只要鼠标不得分六十年代, 它应该接受三试验)。</li>
</ol>3. Rotarod 测试
注: 在 RT 上的杆的表面和直径可能会影响性能。在 RT 上, 如果一只老鼠在第一个十年代的第一个2收集时间点内脱落, 就把它立即放回标尺上进行第二次尝试。
注意: 建议在一天的同一时间运行这些测试, 因为生理和生物化学参数在一天中波动。体重可能影响这项试验的表现, 因此必须加以考虑。
<ol><li>打开理事会 rotarod 和计算机。确保它们已正确连接以进行数据记录。</li>
	<li>确保程序正在运行加速协议。将 rotarod 测试的所需长度设置为 xxx (有效范围为000到 999);在这种情况下, 按 300 *。设置用于 xxx 的车道数 (有效范围为001到 005);在这种情况下, 按 005 *。将起始速度设置为 xxx rpm (输入3005作为 5 rpm; 有效范围为001到 045);在这种情况下, 按 004 *。</li>
	<li>将最高速度设置为 xxx rpm (输入4030为 30 rpm; 有效范围为001到 045);在这种情况下, 按 040 *。将坡道速度设置为 xxx (输入5030以使30秒的坡道进入最高速度; 有效范围为000到 999);在这种情况下, 按 300 *。将单元设置为鼠标模式 (输入6001用于鼠标前进, 6002 用于鼠标反转);在这种情况下, 按 6001 *。</li>
	<li>在开始试验之前, 确保所有磁化车道都是平衡的。将一只老鼠放在 rotarod 的每条车道上。</li>
	<li>按 "开始" 启动程序。当老鼠倒下时, 立即把它们从车道上移开, 以确保它们不会 "绊倒" 其他车道。</li>
	<li>允许老鼠休息10分钟的间隔试验时间。这是1审判的结束。在试验间隔时间内, 擦拭设备, 开始试验下一组小鼠。</li>
	<li>根据需要重复步骤 4-6。如果鼠标停留在 rotarod 上进行完整的5分钟试验, 则不需要对该鼠标进行进一步的试验。</li>
</ol>

<ol>
	<li>Osmon, K. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systemic+Gene+Transfer+of+a+Hexosaminidase+Variant+Using+an+scAAV9.47+Vector+Corrects+GM2+Gangliosidosis+in+Sandhoff+Mice.">Systemic Gene Transfer of a Hexosaminidase Variant Using an scAAV9.47 Vector Corrects GM2 Gangliosidosis in Sandhoff Mice.</a> Human Gene Therapy. 27 (7), 497-508 (2016).</li><li>Brooks, S. P., Dunnett, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tests+to+assess+motor+phenotype+in+mice:+a+user's+guide.">Tests to assess motor phenotype in mice: a user's guide.</a> Nature Reviews, Neuroscience. 10, 519-529 (2009).</li><li>Ma, M., Basso, D. M., Walters, P., Stokes, B. T., Moser, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioural+and+histological+outcomes+following+graded+spinal+cord+contusion+injury+in+the+C57Bl/6+mouse.">Behavioural and histological outcomes following graded spinal cord contusion injury in the C57Bl/6 mouse.</a> Exp. Neurol. 169, 239-254 (2001).</li><li>Menalled, L. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+motor+dysfunction+and+striosomal+distribution+of+huntingtin+microaggregates+in+Huntington's+disease+knock-in+mice.">Early motor dysfunction and striosomal distribution of huntingtin microaggregates in Huntington's disease knock-in mice.</a> J.Neurosci. 22, 8266-8276 (2002).</li><li>Grusser, C., Grusser-Cornehls, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improvement+in+motor+performance+of+Weaver+mutant+mice+following+lesion+of+the+cerebellum.">Improvement in motor performance of Weaver mutant mice following lesion of the cerebellum.</a> Behav. Brain Res. 97, 189-194 (1998).</li><li>De Leonibus, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+deficits+in+a+mouse+model+of+Parkinson's+disease.">Spatial deficits in a mouse model of Parkinson's disease.</a> Psychopharmacology (Berl). 194, 517-525 (2007).</li><li>Morris, M., Koyama, A., Masliah, E., Mucke, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tau+Reduction+Does+Not+Prevent+Motor+Deficits+in+Two+Mouse+Models+of+Parkinson's+Disease.">Tau Reduction Does Not Prevent Motor Deficits in Two Mouse Models of Parkinson's Disease.</a> PLoS ONE. 6 (12), 1-7 (2011).</li><li>Abo-ouf, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deletion+of+tumour+necrosis+factor-&#945;+ameliorates+neurodegeneration+in+Sandhoff+disease+mice.">Deletion of tumour necrosis factor-&#945; ameliorates neurodegeneration in Sandhoff disease mice.</a> Human Molecular Genetics. , 1-16 (2013).</li><li>Dumont, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+Phenotyping+of+Mouse+Models+of+Neurodegeneration.">Behavioral Phenotyping of Mouse Models of Neurodegeneration.</a> Neurodegeneration : Methods and Protocols, Methods in Molecular Biology. 793, 229-237 (2011).</li><li>Cabe, P. A., Tilson, H. A., Mitchell, C. L., Dennis, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+recording+grip+strength+device.">A simple recording grip strength device.</a> Pharmacol. Biochem. 8, 101-102 (1978).</li><li>Barn&#275;oud, P., Loliver, J., Sanger, D., Moser, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+motor+assessment+in+FALS+mice:+a+longitudinal+study.">Quantitative motor assessment in FALS mice: a longitudinal study.</a> Neuroreport. 8, 2861-2865 (1997).</li><li>Lui, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+model+of+GM2+activator+deficiency+manifests+cerebellar+pathology+and+motor+impairment.">Mouse model of GM2 activator deficiency manifests cerebellar pathology and motor impairment.</a> Proc. Natl. Acad. Sci. 94, 8133-8143 (1997).</li><li>Sango, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+Tay-Sachs+and+Sandhoff+diseases+differ+in+neurologic+phenotype+and+ganglioside+metabolism.">Mouse models of Tay-Sachs and Sandhoff diseases differ in neurologic phenotype and ganglioside metabolism.</a> Nature Genetics. 11 (2), 170-176 (1995).</li><li>Jones, B. J., Roberts, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rotarod+suitable+for+quantitative+measurements+of+motor+inco-ordination+in+na&#239;ve+mice.">A rotarod suitable for quantitative measurements of motor inco-ordination in na&#239;ve mice.</a> Nauyn-Schmiedebergs Arch. Pharmacol. 259 (2), 211 (1968).</li><li>Deacon, R. 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=Measuring+the+Strength+of+Mice.">Measuring the Strength of Mice.</a> Journal of Visualized Experiments JoVE. (76), (2013).</li></ol>

作者没有什么可透露的。

对所有的测试, 清洁和消毒的表面, 以尽量减少感染和干扰的小鼠。同时, 在使用经常、RR 和 MT 时, 一定要让老鼠在试验之间和行为测试之间有足够的休息时间。
在经常的情况下, 关键的是每个鼠标都放在开放字段的中间, 并在鼠标释放到中心时启动程序。确保没有手的动作, 传感器可以注册后启动程序。
对于 MT, 重要的是有一个屏障之间的网格区域的鼠标?...

行为测试的目的是测量表型特征, 无论是研究新的小鼠模型的疾病影响中枢神经系统或试用新的治疗。有各种各样的行为测试可供选择。使用行为测试组合的基本原理通常是充分评估临床治疗前的结果。行为测试的电池包括开放现场测试 (经常), 网格测试 (MT) 和 rotarod 测试 (RR)。
常用于一般运动活动的测量1,2,3,4,5,6,7。此评估允许鼠标在一个开放的领域自由移动, 并采取的测量是鼠标在行为试验过程中的动作。使用一个经常包括饲养, 距离移动, 时间花费移动 (例如,行走和跑步) 和活动随着时间的变化 (例如,跳跃和慢/多动运动) 的定量分析, 这就是为什么它是一个有用的和彻底分析啮齿动物的行为。
MT 用于评估鼠标的强度1,2,8,910。使用所有的四肢体挂断铁丝网网格允许非侵入性测量的能力的小鼠表现出持续的肢体张力, 而反对的影响重力。这一评估是通过把鼠标放在一个倒置的网格上, 并定时它的滞后时间下降。这项测试通过测量在脱落前它能在倒置的屏幕上保持多久来评估鼠标的肌肉强度。
RR在使用加速协议1、2、7、8、9、1213中评估鼠标的协调性。rotarod 装置由一个圆柱形光束组成, 在空气中旋转 4 rpm。鼠标可以学习这种运动协调的技能, 然后由缓慢, 增量增加到 40 rpm 挑战。经典的, 恒定速度协议是衡量肌肉力量, 而加速的协议为 RR 评估协调, 耐力和肌肉力量1,2,7,14。
这三行为测试的优点是, 他们被广泛接受的整个啮齿动物行为测试社区2,7,8,9, 当采取不只是作为总和个别测试, 但总的来说, 他们可以被用来作为一个全球评估的表型行为时, 调查不同的结果, 实验治疗。经常是唯一的测试为整体运动活动2。RR 也是评估协调2的唯一方法之一, 而不是使用步态分析软件, 这在这里不会讨论。有几种方法可以评估老鼠的肌肉强度;然而, 使用 MT, 或线挂测试, 已被普遍使用的文献和被接受为可靠的方法测试肌肉力量2,8,9,10,11. 测量一般运动和协调的许多其他技术非常耗时, 可能不会产生可靠的数据 (例如,手动定时鼠标移动或手动绘制用于步态分析的爪子)。自动系统的经常和 RR 增加了数据的可靠性和有效性, 并节省了时间和精力。
例如, 考虑到山德霍夫病小鼠模型的神经, 在执行这一测试电池时, 经常会是这些测试的第一个, 用来评估通常在3月大的时候从这个小鼠模型中看到的运动异常。在12周的年龄, 运动协调方面的赤字也应该开始显现。肌肉力量的缺陷应该在14周的年龄开始, 但最终会比16周更重要。
这些测试是有用的, 当研究导致重大运动损伤的疾病, 包括神经退行性疾病9如山德霍夫疾病1,8,13, 帕金森病7,亨廷顿氏病多发性硬化症这些测试还可以用来研究衰老, 并进行比较的老年评估或健康测量。步态紊乱和运动和平衡的变化在许多这些疾病状态中被看到。然而, 这些测试的用处并不局限于有普遍神经肌肉症状的疾病。
请注意, 在动机或任何被描述的行为测试措施的新奇性方面, 都没有发现混杂变量。在老鼠显示出任何表型症状之前, 研究者获得初步的行为数据是某些疾病状态和病症的必要条件;研究员应该周期性地 (每周) 保证所有马达损伤被交代。许多疾病有快速出现的表型症状, 没有每周的测试, 这些症状不会被占。

<table border="0" cellpadding="0" cellspacing="0" width="629">
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			<td height="21" style="height:21px;width:216px;">IITC Life Sciences Rotarod with Series 8 computing software&#160;</td>
			<td style="width:127px;">IITC Life Sciences</td>
			<td align="right" style="width:119px;">755</td>
			<td style="width:167px;"></td>
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			<td height="42" style="height:42px;">Square Open Field 45 x 45 x 40 cm (grey color). Other floor color available: white, black.</td>
			<td style="width:127px;">Harvard Aparatus (PanLab)</td>
			<td align="right">760190</td>
			<td></td>
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		<tr height="42">
			<td height="42" style="height:42px;">&#160;Open Field Preconfigured Module</td>
			<td style="width:127px;">Harvard Aparatus (PanLab)</td>
			<td align="right">760688</td>
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			<td height="42" style="height:42px;">Smart V3.0 Software Platform&#160;</td>
			<td style="width:127px;">Harvard Aparatus (PanLab)</td>
			<td align="right">760681</td>
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			<td height="42" style="height:42px;">Video converter (analog/digital)</td>
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			<td align="right">760262</td>
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			<td height="42" style="height:42px;">Roof fixing sys for camera</td>
			<td style="width:127px;">Harvard Aparatus (PanLab)</td>
			<td align="right">760277</td>
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battery of behavioral tests assessing general locomotion, muscular strength, and coordination in mice

行为测试用于临床前试验, 以评估特定疾病或治疗对动物的福祉和健康的表型效应和结果。有许多行为测试可以应用。我们选择了一般运动测试, 野外测试 (经常);肌肉强度的测试, 网格测试 (MT);和测试的协调, rotarod 测试 (RR)。测试可以在每周或每月的基础上完成。作为一般运动的一个测试, 经常工作通过客观地监测运动参数, 而鼠标是在野外设备。该领域通常是一个 2 ' x 2 ' 盒, 并通过激光传感或视频捕获记录的运动。鼠标放在开放字段的中心, 允许自由移动以进行测试。MT 使用鼠标从一个倒置的屏幕上掉下来作为衡量肌肉力量的时间。将鼠标放在屏幕上, 在一个清晰的框上反转, 并定时等待它们的滞后时间下降。进行了三项试验, 最好的三项试验是在当天取得的。六十年代的分数是鼠标左倒的最长时间。在网格测试试验中, 给老鼠5分钟的休息时间。最后, 关于 RR 的加速的协议评估马达协调和耐力。在试验中, 鼠标在旋转杆上行走, 因为它在转速从 4 rpm 增加到 40 rpm 超过5分钟。试验结束时, 鼠标接触磁化压力传感器后下降。每只老鼠经历了三次试验, 最好的试验是在那一天取得的。这种综合行为数据允许对测试动物的移动性、协调性、强度和运动进行全球评估。在三行为测试措施中, 至少有两项必须表现出对动物的改进, 以使其具有整体改善运动功能的资格。

研究动物可以每周或每月接受行为测试范式, 以适应分析所需的必要时间点。有关行为测试计时的示例, 请参见图 1 。在下面的代表性数据中, 取自 Osmon et al。(2016), 小鼠每月接受行为测试, 从12周开始, 直到他们的人性化终点大约40周龄。行为测试的时间对实验来说有时是至关重要的, 特别是当未经处理的与经过治疗的动物的生存范围很广并且需要进行比?...

Watch this Scientific Journal Video about Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice at JoVE.com

Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

行为测试用于临床试验, 以评估疾病或治疗对动物福祉的表型影响。为了在全球范围内评估电机的功能, 我们选择了一般运动, 肌肉力量和协调的测试: 开放现场测试, 网格测试, 和 rotarod 测试, 分别。

用于评估一般运动、肌肉力量和小鼠协调性的行为测试电池

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the ...

battery-behavioral-tests-assessing-general-locomotion-muscular

Research

JoVE Journal

Behavior

13.5K 次观看.  Queen's University. 行为测试用于临床试验, 以评估疾病或治疗对动物福祉的表型影响。为了在全球范围内评估电机的功能, 我们选择了一般运动, 肌肉力量和协调的测试: 开放现场测试, 网格测试, 和 rotarod 测试, 分别。

视频: Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

Shallow Water (Paddling) Variants of Water Maze Tests in Mice

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to the mouse becoming increasingly important. But researchers found that some strains of mutant mice were prone to problems like passively floating or diving when they were tested in the Morris water maze11. This was unsurprising considering their natural habitat; rats swim naturally (classically, the "sewer rat"), whereas mice evolved in the dry areas of central Asia.
To overcome these problems, it was considered whether shallow water would be a sufficient stimulus to provide escape motivation for mice. This would also avoid the problems of drying the small creatures with a towel and then putting them in a heated recovery chamber to avoid hypothermia, which is a much more serious problem than with rats; the large ratio of surface area to volume of a mouse makes it particularly vulnerable to rapid heat loss.
Another consideration was whether a more natural escape strategy could be used, to facilitate learning. Since animals that fall into water and swim away from the safety of the shore are unlikely to pass on their genes, animals have evolved a natural tendency to swim to the edge of a body of water. The Morris water maze, however, requires them to swim to a hidden platform towards the center of the maze - exactly opposite to their evolved behavior. Therefore the paddling maze should incorporate escape to the edge of the apparatus. This feature, coupled with the use of relatively non-aversive shallow water, embodies the "Refinement" aspect of the "3 Rs" of Russell and Burch8.
Various types of maze design were tried; the common feature was that the water was always shallow (2 cm deep) and escape was via a tube piercing the transparent wall of the apparatus. Other tubes ("false exits") were also placed around the walls but these were blocked off. From the inside of the maze all false exits and the single true exit looked the same. Currently a dodecagonal (12-sided) maze is in use in Oxford, with 12 true/false exits set in the corners. In a recent development a transparent paddling Y-maze has been tested successfully.

Shallow Water Paddling Variants of Water Maze Tests in Mice

Mice can swim, but many strains appear to find this activity stressful. To overcome this problem mazes have been devised where escape from shallow water is used to motivate behaviour. These have been demonstrated to support learning at least as good as the traditional and widely used Morris water maze.

浅水（杖刑）的变体在小鼠体内的水迷宫测试

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to ...

科研

行为学

Marble Burying and Nestlet Shredding as Tests of Repetitive, Compulsive-like Behaviors in Mice

Obsessive-compulsive disorder (OCD) and autism spectrum disorders (ASD) are serious and debilitating psychiatric conditions and each constitutes a significant public health concern, particularly in children. Both of these conditions are highlighted by the repeated expression of meaningless behaviors. Individuals with OCD often show checking, frequent hand washing, and counting. Children with ASDs also engage in repetitive tapping, arm or hand flapping, and rocking. These behaviors can vary widely in intensity and frequency of expression. More intense forms of repetitive behaviors can even result in injury (e.g.&#160;excessive grooming, hand washing, and self-stimulation). These behaviors are therefore very disruptive and make normal social discourse difficult. Treatment options for repetitive behaviors in OCD and ASDs are somewhat limited and there is great interest in developing more effective therapies for each condition. Numerous animal models for evaluating compulsive-like behaviors have been developed over the past three decades. Perhaps the animal models with the greatest validity and ease of use are the marble burying test and the nestlet shredding test. Both tests take advantage of the fact that the target behaviors occur spontaneously in mice. In the marble burying test, 20 marbles are arrayed on the surface of clean bedding. The number of marbles buried in a 30 min session is scored by investigators blind to the treatment or status of the subjects. In the nestlet shredding test, a nestlet comprised of pulped cotton fiber is preweighed and placed on top of cage bedding and the amount of the nestlet remaining intact after a 30 min test session is determined. Presently, we describe protocols for and show movie documentation of marble burying and nestlet shredding. Both tests are easily and accurately scored and each is sensitive to small changes in the expression of compulsive-like behaviors that result from genetic manipulations, disease, or head injury.

Disease, head injury, genetic modifications, and treatment of mice with drugs can have profound effects on behavior. Utilizing well-characterized and validated approaches such as marble burying and nestlet shredding, compulsive-like behaviors can be documented accurately in mice as models of human obsessive-compulsive disorder and autism spectrum disorder.

Obsessive-compulsive disorder (OCD) and autism spectrum disorders (ASD) are serious and debilitating psychiatric conditions and each constitutes a ...

Operant Procedures for Assessing Behavioral Flexibility in Rats

Executive functions consist of multiple high-level cognitive processes that drive rule generation and behavioral selection. An emergent property of these processes is the ability to adjust behavior in response to changes in one&#8217;s environment (i.e., behavioral flexibility). These processes are essential to normal human behavior, and may be disrupted in diverse neuropsychiatric conditions, including schizophrenia, alcoholism, depression, stroke, and Alzheimer&#8217;s disease. Understanding of the neurobiology of executive functions has been greatly advanced by the availability of animal tasks for assessing discrete components of behavioral flexibility, particularly strategy shifting and reversal learning. While several types of tasks have been developed, most are non-automated, labor intensive, and allow testing of only one animal at a time. The recent development of automated, operant-based tasks for assessing behavioral flexibility streamlines testing, standardizes stimulus presentation and data recording, and dramatically improves throughput. Here, we describe automated strategy shifting and reversal tasks, using operant chambers controlled by custom written software programs. Using these tasks, we have shown that the medial prefrontal cortex governs strategy shifting but not reversal learning in the rat, similar to the dissociation observed in humans. Moreover, animals with a neonatal hippocampal lesion, a neurodevelopmental model of schizophrenia, are selectively impaired on the strategy shifting task but not the reversal task. The strategy shifting task also allows the identification of separate types of performance errors, each of which is attributable to distinct neural substrates. The availability of these automated tasks, and the evidence supporting the dissociable contributions of separate prefrontal areas, makes them particularly well-suited assays for the investigation of basic neurobiological processes as well as drug discovery and screening in disease models.

The ability to assess executive functions such as behavioral flexibility in rats is useful for investigating the neurobiology of cognition in both intact animals and disease models. Here we describe automated tasks for assessing strategy shifting and reversal learning, which are particularly sensitive to disruptions in prefrontal cortical networks.

操作性程序，以评估大鼠的行为灵活性

Executive functions consist of multiple high-level cognitive processes that drive rule generation and behavioral selection. An emergent property of these ...

Asymmetric Walkway: A Novel Behavioral Assay for Studying Asymmetric Locomotion

Behavioral assays are commonly used for the assessment of sensorimotor impairment in the central nervous system (CNS). The most sophisticated methods for quantifying locomotor deficits in rodents is to measure minute disturbances of unconstrained gait overground (e.g., manual BBB score or automated CatWalk). However, cortical inputs are not required for the generation of basic locomotion produced by the spinal central pattern generator (CPG). Thus, unconstrained walking tasks test locomotor deficits due to motor cortical impairment only indirectly. In this study, we propose a novel, precise foot-placement locomotor task that evaluates cortical inputs to the spinal CPG. An instrumented peg-way was used to impose symmetrical and asymmetrical locomotor tasks mimicking lateralized movement deficits. We demonstrate that shifts from equidistant inter-stride lengths of 20% produce changes in the forelimb stance phase characteristics during locomotion with preferred stride length. Furthermore, we propose that the asymmetric walkway allows for measurements of behavioral outcomes produced by cortical control signals. These measures are relevant for the assessment of impairment after cortical damage.

Here, we present a protocol to quantify precise stepping in rodents. Cortical and the spinal central pattern generator signals are required for precise foot-placement during obstructed locomotion. We report here the novel constrained walking task that directly examines precise stepping behavior.

非对称走道:一种新的行为检测方法的研究不对称步态

Behavioral assays are commonly used for the assessment of sensorimotor impairment in the central nervous system (CNS). The most sophisticated methods for ...

Assessment of Cocaine-induced Behavioral Sensitization and Conditioned Place Preference in Mice

It is thought that rewarding experiences with drugs create strong contextual associations and encourage repeated intake. In turn, repeated exposures to drugs of abuse make lasting alterations in the brain function of vulnerable individuals, and these persistent alterations likely serve to maintain the maladaptive drug seeking and taking behaviors characteristic of addiction/dependence2. In rodents, reward experience and contextual associations are frequently measured using the conditioned place preference assay, or CPP, wherein preference for a previously drug-paired context is measured. Behavioral sensitization, on the other hand, is an increase in a drug-induced behavior that develops progressively over repeated exposures. Since sensitized behaviors can often be measured after several months of drug abstinence, depending on the dose and length of initial exposure, they are considered observable correlates of lasting drug-induced plasticity. Researchers have found these assays useful in determining the neurobiological substrates mediating aspects of addiction as well as assessing the potential of different interventions in disrupting these behaviors. This manuscript describes basic, effective protocols for mouse CPP and locomotor behavioral sensitization to cocaine.

This protocol is intended to enable researchers to conduct experiments designed to test these aspects of addiction using the conditioned place preference and locomotor behavioral sensitization assays.

可卡因诱导行为敏和小鼠位置偏爱评估

It is thought that rewarding experiences with drugs create strong contextual associations and encourage repeated intake. In turn, repeated exposures to ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

鼠类精神运动警觉测试（rPVT）:一个在大鼠和小鼠的神经行为评估性能的方法

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

Assessing Working Memory in Children: The Comprehensive Assessment Battery for Children &#8211; Working Memory (CABC-WM)

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of working memory in young school-age children. Working memory deficits have been identified in children with language-based learning disabilities, including dyslexia1,2 and language impairment3,4, but it is not clear whether these children exhibit deficits in subcomponents of working memory, such as visuospatial or phonological working memory. The CABC-WM is administered on a desktop computer with a touchscreen interface and was specifically developed to be engaging and motivating for children. Although the long-term goal of the CABC-WM is to provide individualized working memory profiles in children, the present study focuses on the initial success and utility of the CABC-WM for measuring central executive, visuospatial, phonological loop, and binding constructs in children with typical development. Immediate next steps are to administer the CABC-WM to children with specific language impairment, dyslexia, and comorbid specific language impairment and dyslexia.

Working memory predicts a significant amount of variance for a variety of cognitive tasks, including speaking, reading, and writing. However, few tools are available to assess working memory in children. We present an innovative, computer-based battery that comprehensively assesses different components of working memory in school-age children.

评估儿童工作记忆:儿童综合评估电池 - 工作记忆（CABC-WM）

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of ...

Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia

In keeping with the 3R Principle (Replacement, Reduction, Refinement) developed by Russel and Burch, scientific research should use alternatives to animal experimentation whenever possible. When there is no alternative to animal experimentation, the total number of laboratory animals used should be the minimum needed to obtain valuable data. Moreover, appropriate refinement measures should be applied to minimize pain, suffering, and distress accompanying the experimental procedure. The categories used to classify the degree of pain, suffering, and distress are non-recovery, mild, moderate, or severe (EU Directive 2010/63). To determine which categories apply in individual cases, it is crucial to use scientifically sound tools.
The well-being-assessment protocol presented here is designed for procedures during which general anesthesia is used. The protocol focuses on home cage activity, the Mouse Grimace Scale, and luxury behaviors such as burrowing and nest building behavior as indicators of well-being. It also uses the free exploratory paradigm for trait anxiety-related behavior. Fecal corticosterone metabolites as indicators of acute stress are measured over the 24-h post-anesthetic period.
The protocol provides scientifically solid information on the well-being of mice following general anesthesia. Due to its simplicity, the protocol can easily be adapted and integrated in a planned study. Although it does not provide a scale to classify distress in categories according to the EU Directive 2010/63, it can help researchers estimate the degree of severity of a procedure using scientifically sound data. It provides a way to improve the assessment of well-being in a scientific, animal-centered manner.

We developed a protocol to assess well-being in mice during procedures using general anesthesia. A series of behavioral parameters indicating levels of well-being as well as glucocorticoid metabolites were analyzed. The protocol can serve as a general aid to estimate the degree of severity in a scientific, animal-centered manner.

全身麻醉下小鼠幸福度的系统评价

In keeping with the 3R Principle (Replacement, Reduction, Refinement) developed by Russel and Burch, scientific research should use alternatives to animal ...

Assessing Activity-based Anorexia in Mice

Rodents develop activity-based anorexia (ABA) when exposed to a restricted feeding schedule and allowed free access to a running wheel. These conditions lead to a life-threatening reduction in body weight. However, rodents exposed to only one of these conditions ultimately adapt to re-establish normal body weight. Although increased running coupled with reduction in voluntary food intake appear paradoxical under ABA conditions, ABA behavior is observed across numerous mammalian species. 
The ABA paradigm provides an animal model for anorexia nervosa (AN), an eating disorder with severe dysregulation of appetite-behavior. Subjects are singly housed with free access to a running wheel. Each day, the subject is offered food for a limited amount of time. During the course of the experiment, a subject's body weight decreases from high activity and low caloric intake. The duration of the study varies based on how long food is offered daily, the type of food offered, the strain of mouse, if drugs are being tested, and environmental factors.
A lack of effective pharmacological treatments for AN patients, their low quality of life, high cost of treatment, and their high mortality rate indicate the urgency to further research AN. We provide a basic outline for performing ABA experiments with mice, offering a method to investigate AN-like behavior in order to develop novel therapies. This protocol is optimized for use in Balb/cJ mice, but can easily be manipulated for other strains, providing great flexibility in working with different questions, especially related to genetic factors of ABA.

Mice individually housed with a running wheel while given limited access to food develop reductions in food consumption and increase activity on the running wheel. This experimental phenomenon is called activity-based anorexia. This paradigm provides an experimental tool for studying the neurobiology and behaviors underlying aspects of anorexia nervosa.

小鼠活动性厌食症的评估

Rodents develop activity-based anorexia (ABA) when exposed to a restricted feeding schedule and allowed free access to a running wheel. These conditions ...

The Unpredictable Chronic Mild Stress Protocol for Inducing Anhedonia in Mice

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by many patients prompts the need to develop new therapeutic alternatives and to better understand the etiology of the disorder. Pre-clinical models with translational merits are rudimentary for this task. Here we present a protocol for the unpredictable chronic mild stress (UCMS) method in mice. In this protocol, adolescent mice are chronically exposed to interchanging unpredictable mild stressors. Resembling the pathogenesis of depression in humans, stress exposure during the sensitive period of mice adolescence instigates a depressive-like phenotype evident in adulthood. UCMS can be used for screenings of antidepressants on the variety of depressive-like behaviors and neuromolecular indices. Among the more prominent tests to assess depressive-like behavior in rodents is the sucrose preference test (SPT), which reflects anhedonia (core symptom of depression). The SPT will also be presented in this protocol. The ability of UCMS to induce anhedonia, instigate long-term behavioral deficits and enable reversal of these deficits via chronic (but not acute) treatment with antidepressants strengthens the protocol&#39;s validity compared to other animal protocols for inducing depressive-like behaviors.

Here we present the unpredictable chronic mild stress protocol in mice. This protocol induces a long-term depressive-like phenotype and enables to assess the efficacy of putative antidepressants in reversing the behavioral and neuromolecular depressive-like deficits.

诱导小鼠快感缺失的不可预测慢性轻度应力协议

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by ...