The authors would like to thank Theresa Jones, Ph.D. for her guidance and assistance in adapting the reaching task to mice. Data presented in this manuscript were supported by grants from NIH-NINDS (NS64586 to TAJ and NS076275 to ALK) and a predoctoral NRSA to KAT (F31AG034032). The NIH was not involved in any aspect of study designs or analyses presented in this manuscript nor in the composition of this manuscript.

下面的方法是按照批准的得克萨斯大学奥斯汀分校和伊利诺伊卫斯理动物护理和使用委员会的协议。建议研究人员请戴上手套或在行为训练与任何实验动物从事时采取预防措施（前，后洗手）。在准备和手术过程中处理动物时，必须戴好手套。 1，习惯化和食品的限制注：由于PMRT是​​一个食欲的任务，这是有利于限制饲喂的训练开始之前，以激励深远反应，并防止在培训期间饱足感的金额及时间。 <ol><li>小鼠放置在限饲日程安排至少1周任务训练开始之前开始。称量每只小鼠的食物限制发病前建立自由摄食的权重，其中被确定为次每个动物在限饲的第一天电子权值。 <ol><li>每日一次喂小鼠，每个鼠标接收约2.5克，每天标准啮齿动物食物（ 例如 ，如果有4只小鼠放置在一个笼子里，笼子将每天获得10克食物）。确保喂养每天发生在同一时间，最好在当时每天训练有望结束。 </li><li>经常称一称体重的动物，以确保没有鼠标失去他们的自由摄食量的10％以上。注：调整日常食品金额相应地促进适当维持体重。老鼠通常体重增加，因为他们成熟，甚至与恒限食鼠留在食物限制PMRT培训/考核的持续时间。 </li></ol></li><li>暴露动物至更小的片的面食（切至1.6厘米），其家笼几次之前的训练，以克服恐新的反应。注：通常5-10天的接触是足够的，以每只小鼠普罗维约4个副执行干事在每次喂奶。 </li><li>用剃刀刀片切面食片尺寸（3.2公分）。注意：建议在过量的面食片前开始一天的训练，并存储在一个密封的容器未切件。卡佩利尼面食来填充矩阵。作为不同品牌的面食变化在质地和直径，这可能会影响面食在基质或所需的力的量为小鼠破面食的稳定性，所以建议DeCecco品牌面食在整个训练和测试使用。 </li><li>将未煮过的卡佩利尼面食片的面食基质，填充矩阵中所有可用的孔（ 图1A）。 </li><li>习惯于小鼠在试验室中采用了全面食基质放置在前方（ 图1A）和几片的面食吃在测试腔室的底部。习惯于老鼠5-10分钟3次。注：小鼠不应该鼓励habitua期间达成的面食化训练。鼠标通常开始出现在位于深远室外面由它通过光圈嗅探面食兴趣。这是可能的，如果需要的习惯于2笼配合在一个单一的腔室，但小鼠的形状应与单独训练。所有表面应当用70％乙醇或类似的，安全的消毒液进行清洗之前和之后每个动物被放置在装置的行为。 </li></ol> 2，整形河段<ol><li>通过塑造达到了3-5每日一次成型试验确定每个动物的首选肢体。允许小鼠达到10分钟，或者总共10次，其中第一次出现。 <ol><li>放置老鼠分别在测试腔与所述腔室的前端的完整的面食矩阵作为训练习惯（ 图1A）。 </li><li>记录达到的数目与每个肢体，分别表示与右下游和左肢的数目。定义一个范围的扩展通过接触孔，使得腕分解室的平面内的肢体;这是没有必要的范围也可以朝向或作出与面条的接触。 </li></ol></li><li>通过光圈达到保持几件面食室只是里面鼓励的老鼠都不愿意接触。绘制面食片出来，当鼠标显示的兴趣（ 如嗅探，咬），从而鼓励他们到达撤退面食片之后。如果鼠标没有最初显示在干面食兴趣，那么湿面食枚一小杯温水的技巧，使他们更可口。 </li><li>确定肢体的偏好，当鼠标表现出最低的日常达到70％与一个单一的肢体。不要过度形状的动物，因为这样可以限制学习的达成任务17培训部分期间观察到的金额。开始PMRT培训，一旦老鼠的形状。注：培训可以尽快NEX启动吨天，但也是可以接受的等待整形和训练程序之间的数天。 </li></ol> 3，到达训练<ol><li>火车小鼠每日一次之前，侮辱，以建立能力上的PMRT。注：每个审判由最多15分钟或100到达观察员（成功的和不成功的）计算，以先到为准的。老鼠被训练来达到只与自己喜欢的肢体。老鼠会容忍每天两次试验，如果一个更紧张的训练模式需要18。 <ol><li>放置老鼠在测试腔室与一个半满的矩阵放置在前方（ 图1B）。火车小鼠只与自己喜欢的肢体发消息对侧矩阵只有一侧的肢体首选，因为老鼠在他们的身体中线到达到达。留下对应于不优选肢基质未填充，以阻止到达尝试与该肢（这将不会是S的部uccessful因为他们无法达到面食），并鼓励只用首选肢体接触。 </li></ol></li><li>将少量的压力，面食，而它在矩阵，以帮助老鼠在早期的培训课程成功突破件。不破面食片的鼠标。通过保持一个或两个面食片轻轻后面并垂直于预期的到达目标实现这一点。这些少量​​的支持有助于稳定薄面食，这是相当灵活的，困难的小鼠把握，因为他们最初的学习任务。 </li><li>记录达到，成功达到的数目，并成功地达到的位置的总数。成功达到要求的鼠标，达到通过光圈，抢了一块面食，并打破意大利面片从基质中取出。注：面食件不应取代矩阵中，直到审判结束之后。早在训练，老鼠不会打破许多面食片。 </li><li>列车小鼠在PMRT到熟练程度，其定义为一致的性能至少3天（和5天）（天以上断裂至少9个面食具有不超过2个更多或更少个）。它通常需要15-20天之间，以培养年轻的成年小鼠的熟练程度。 </li></ol> 4，受影响较小的肢体训练<ol><li>训练受影响较小的肢体（ 即手术前不可取的肢体）开始缺血性损伤的运动皮层对侧首选达到肢体四天后（参见图2为澄清受影响/首选和less-affected/unpreferred四肢） 。管理每日一次15分钟或100到达的培训课程。 <ol><li>删除面食的第一列（ 即 ，从到达孔延伸，最内侧的垂直列）受影响较小的肢体训练的第一周，以确保与早期患侧肢体到达都没有成功。 </li><li&gt;放置老鼠与半满矩阵放置在前方的到达腔室。确保填充矩阵的一半是在手术前填充一半，这将迫使老鼠到现在与受影响较小的（不优选）肢到达的对面。 </li></ol></li><li>鼓励与受影响较小的肢体透过光圈达到增强扩展这个肢体接触。当鼠标延长受影响较小的肢体训练到位的最初几天的小（1/2片）面食到到达室的地板无论接触不到的地方是否成功。 </li><li>只计算那些与受影响较小的肢体朝至100范围总做河段。不要指望鬼鬼祟祟的尝试与对达到总数患肢。 </li><li>返回面食的最内侧竖列的矩阵中的第7天受影响较小的肢体训练后。 </li><li>记录达到，成功达到的数目，和SUCC的位置的数量essful河段。注意：这些数据可以被分析以观察在到达模式与较少受影响肢体的差异，但还没有报道。 </li></ol> 5，到达分析确定通过评估对PMRT任务如下受影响较小的肢体训练患侧肢体上的患肢功能恢复受影响较小的肢体运用的影响。发生这种评估后的欠患侧肢体训练天所需的数字是否齐全（一般14天）。 <ol><li>将基质在前面商会，面向相同的训练程序（ 即 ，与基体对侧的肢体首选的填充一半）。注意：这迫使小鼠达到他们的影响（和以前优先）肢体。 </li><li>记录达到，成功达到的数目，并成功地达到的位置与预先冲程的性能水平进行比较的总数。不要探测性能在受影响较小的肢体训练，这些受影响的肢体探头患肢可作为培训课程，可能会影响患肢的恢复。 </li></ol>

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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+and+neuroplastic+effects+of+focal+endothelin-1+induced+sensorimotor+cortex+lesions.">Behavioral and neuroplastic effects of focal endothelin-1 induced sensorimotor cortex lesions.</a> Neuroscience. 128 (3), 473-486 (2004).</li><li>Bryant, A., Bernier, B., Jones, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Abnormalities+in+skilled+reaching+movements+are+improved+by+peripheral+anesthetization+of+the+less-affected+forelimb+after+sensorimotor+cortical+infarcts+in+rats.">Abnormalities in skilled reaching movements are improved by peripheral anesthetization of the less-affected forelimb after sensorimotor cortical infarcts in rats.</a> Behav. Brain Res. 177 (2), 298-307 (2007).</li><li>Whishaw, I. Q., Coles, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Varieties+of+paw+and+digit+movement+during+spontaneous+food+handling+in+rats:+Postures,+bimanual+coordination,+preferences,+and+the+effect+of+forelimb+cortex+lesions.">Varieties of paw and digit movement during spontaneous food handling in rats: Postures, bimanual coordination, preferences, and the effect of forelimb cortex lesions.</a> Behav. Brain Res. 77 (1-2), 135-148 (1996).</li><li>Whishaw, I. Q., Dringenberg, H. C., Pellis, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spontaneous+Forelimb+Grasping+in+Free+Feeding+by+Rats+-+Motor+Cortex+Aids+Limb+and+Digit+Positioning.">Spontaneous Forelimb Grasping in Free Feeding by Rats - Motor Cortex Aids Limb and Digit Positioning.</a> Behav. Brain Res. 48 (2), 113-125 (1992).</li><li>Horie, N., Maag, A., Hamilton, S. A., Shichinohe, H., Bliss, T. M., Steinberg, G. K. <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+focal+cerebral+ischemia+using+endothelin-1.">Mouse model of focal cerebral ischemia using endothelin-1.</a> J. Neurosci. Methods. 173 (2), 286-290 (2008).</li><li>Maldonado, M. A., Allred, R. P., Felthauser, E. L., Jones, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+skill+training,+but+not+voluntary+exercise,+improves+skilled+reaching+after+unilateral+ischemic+lesions+of+the+sensorimotor+cortex+in+rats.">Motor skill training, but not voluntary exercise, improves skilled reaching after unilateral ischemic lesions of the sensorimotor cortex in rats.</a> Neurorehabil. Neural Repair. 22 (3), 250-261 (2008).</li><li>Clarkson, A. N., Lopez-Valdes, H. E., Overman, J. J., Charles, A. C., Brennan, K. C., Carmichael, S. 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The authors have nothing to disclose.

该PMRT表示一个简单的，定量的方法来评估小鼠技能达到的性能。虽然费时，有可能以前没有经验的大学生进行培训，以收集可靠和可重复的数据，只有几个培训班。该任务是足够敏感的测量缺血性侮辱3，13，15在小鼠运动技能表现，甚至轻微的变化，以及多项研究引用的长期赤字与各种培训协议15，20，22，因为它是一个单边的任务，它也可以被用来规定肢体与小鼠执行的任务。该...

小鼠模型中已经变得越来越流行用于实验中风研究部分地由于其便利性和经济性，以及转基因株系，适合用于在其他应用中的体内成像的可用性。有了这个日益普及，在实验模型，在开发功能性结果的敏感行为评估损伤后的利息也增加了1-7。动物训练协议，模仿两个康复和补偿策略使用的人中风幸存者的发展提高到成功地从实验动物的研究成果转化的发现在诊所8使用的能力。对面食矩阵运动技能训练晚晴任务（PMRT）先前已确立为运动技能的结果如下感觉运动皮层3的缺血性损伤敏感的行为评估。 其中一个主要的兴趣S IN中风研究关注的康复和行为策略，促进功能恢复改善以下侮辱的发展和理解。目前，在人类的康复策略导致不完全恢复8。此外，康复治疗师必须打击补偿性策略，在恢复期间中风幸存者的发展，可能会破坏他们完全恢复其患肢（S）的功能的能力。例如，下面的单方面中风，影响上肢功能，人类倾向于发展自己的受影响较小的肢体9,10的依赖，同时提高一个人的在短期内发挥作用的能力，这个教训不使用患肢可能妨碍其最终采收率的潜力，这表现在动物模型11-13。这些发现在动物有助于告知约束诱导运动疗法的开发和利用在人类14。动物模型是beneficial对于通过使研究人员能够探索subserve的神经生物学机制，促进功能恢复提高恢复战略。除了 ​​作为卒中后功能的有效的行为评估，PMRT已被确立为一种有效的康复策略，以促进改善功能转归以下感觉运动行程15。该PMRT也可以用来有效地模仿学会不使用患侧肢体，因此，尽管提供了最初的洞察行为的操作，可能会改善功能恢复过度依赖于受影响较小的肢体13。 该PMRT建设之前已经3描述。简单地说，到达腔室是一个开放的顶部和底部由四个有机玻璃墙（20厘米高，长15厘米和8.5厘米宽）。还有一个中心切口（13厘米高和宽5mm）从供应腔室的前壁的底部基座延伸为达到光圈（ 图1A）。面食的矩阵是一个重型塑料块（8.5厘米长，5cm宽，150厘米高），通过该块的深度钻出完全直径为1毫米的孔。共有260孔，从2 mm从该窗口达到每孔（ 图1B）为2毫米。面食矩阵被设计为使得干燥，垂直定向的面食片通过矩阵阶段大约一半的面食片露出的整个深度延伸。一个可移动件的开销塑料或卡片纸应切成一定尺寸和牢固地贴在基体的下侧。这可以防止面食件在运输过程中掉落出来的矩阵，并可以很容易地拆卸破碎面食件。 该PMRT是​​一个多功能的和敏感的行为分析，允许实验者收集准确的结果数据和操作使用肢体模仿的临床现象。作为一种行为Øutcome措施，PMRT允许实验者收集的行为数据可更准确地反映了康复策略并不比传统的测量梗死面积3,16的有效性。作为行为操纵，该PMRT允许实验者来控制在小鼠上肢用在以模仿康复的临床经验（ 即患侧肢体训练）或学习不使用（ 即受影响较小的肢体训练）。当与神经解剖相结合的方法，对PMRT为研究者提供了一个机会，探讨支持的功能或适应不良行为的可塑性后恢复中风后肢体代偿使用的机制。该PMRT可以进一步应用到脑损伤和上肢障碍的其他小鼠模型，如创伤性脑损伤。该PMRT的另一个优点是它的承受能力。任务所需的装备可以在家里还算合理构建，数据收集不并不需要大量的空间，或财务资源，任务是很简单的本科生可靠地采集数据。另外，PMRT是即使小行为缺陷3,13敏感，该协议提供了一种简单而有效的方法，以评估运动技能学习，促进行为恢复损伤后，并在行程的建立小鼠模型模仿了解到非使用的现象。

<table border="0" cellpadding="0" cellspacing="0" width="794">
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			<td height="42" style="height:42px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:71px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:388px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Reaching Chamber</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td>Reaching chambers are made in house with Plexiglas</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Pasta Matrix Block</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td>The Pasta Matrix box is made in house using a heavy plastic block</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Capellini Pasta</td>
			<td style="width:71px;">DeCecco</td>
			<td style="width:119px;"></td>
			<td style="width:388px;">DeCecco brand capellini pasta can be purchased in a grocery store or through an online retailer such as Amazon</td>
		</tr>
	</tbody>
</table>

compensatory limb use and behavioral assessment of motor skill learning following sensorimotor cortex injury in a mouse model of ischemic stroke

小鼠模型已成为越来越受欢迎的行为神经科学领域中，特别是中风的实验研究。作为模型的进步，它制定具体的鼠标敏感行为的措施是很重要的。本协议描述了在中风小鼠模型使用一个熟练的动作任务。该面食矩阵到达任务功能作为一个多功能的和敏感的行为分析，允许实验者收集准确的结果数据和操作使用肢体模仿人体临床现象，包括补偿性策略（ 即 ，学会不使用），重点突出康复训练。当与神经解剖工具相结合，这个任务也允许研究人员能够探索支持的功能（或缺乏）行为学恢复中风后的机制。任务既简单又经济实惠的设立和开展，提供了多种用于有关福无数研究问题的培训和测试选项nctional结果如下伤害。虽然该任务已被应用于中风的小鼠模型，它也可能是有益的，在其他上肢损伤模型的功能结果的研究。

从PMRT分析的结果应包括面食片的数量打破，达到成功的模式。结果从小鼠感觉运动皮质病变表明，缺血性损伤影响成功的达到双方人数以及身体接触模式3，19，这表现在图3A。代表图案的变化可以在图3B观察。该PMRT的模拟小鼠学习非使用效果的能力表现在患肢以下较少受影响的肢体训练13，20的受损的恢复，其结果是，只有成功地达到与受影响的肢体进行?...

Watch this Scientific Journal Video about Compensatory Limb Use and Behavioral Assessment of Motor Skill Learning Following Sensorimotor Cortex Injury in a Mouse Model of Ischemic Stroke at JoVE.com

Compensatory Limb Use and Behavioral Assessment of Motor Skill Learning Following Sensorimotor Cortex Injury in a Mouse Model of Ischemic Stroke

Mouse models have become increasingly popular in studies of behavioral neuroscience. As models advance, it is important to develop sensitive behavioral measures specific to the mouse. This protocol describes the Pasta Matrix Reaching Task, which is a skilled motor task for use in mouse models of stroke.

补偿肢使用和运动技能学习行为评估以下感觉运动皮层损伤缺血性脑卒中的小鼠模型

Mouse models have become increasingly popular in the field of behavioral neuroscience, and specifically in studies of experimental stroke. As models ...

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Behavior

11.3K 次观看.  Illinois Wesleyan University. Mouse models have become increasingly popular in studies of behavioral neuroscience. As models advance, it is important to develop sensitive behavioral measures specific to the mouse. This protocol describes the Pasta Matrix Reaching Task, which is a skilled motor task for use in mouse models of stroke.