Name: the knob supination task: a semi-automated method for assessing forelimb function in rats
Uploaded: 2017-09-28T15:00:00.000+00:00
Duration: 9 min 26 s
Description: Watch this Scientific Journal Video about The Knob Supination Task: A Semi-automated Method for Assessing Forelimb Function in Rats at JoVE.com

这项研究由 NIH-一 R03 NS091737 资助。

该协议描述了设置任务和建立生存和喂养条件, 以及训练动物、损伤后测试动物以及分析行为数据。动物训练的四步也被描述: 适应, 报偿协会, 配重训练和训练到标准转折角度。所有的动物实验都得到了 IACUC 康奈尔医学和德克萨斯大学达拉斯分校的批准. 
 1. 设置任务 <ol> <li> 设计一个由透明塑料制成的行为盒, 它的宽度为 150 mm, 长度为 200 mm, 长 250 mm. 
	注: 在这里, 矩形孔径是14.2 毫米宽25.4 毫米高。直径9.5 毫米的旋钮 manipulandum 是用甲基丙烯酸酯制成的, 有两个停止, 限制旋小于100和 #176;。每个参数都经过测试, 并针对结果部分中描述的缺陷进行了优化。行为框和 manipulandum 显示在 图 1A 中. </li>
	<li> 将微控制器框连接到计算机。多达四个单片机盒可以连接到每台计算机. </li>
	<li> 若要开始任务, 请使用控制软件 ( 图 1C )。
	<ol> <li> 选择要控制的旋钮设备。在 #34 中输入主题名称; 主题和 #34; 字段。选择 #34 下的培训阶段; 舞台和 #34; 下拉菜单. 
		注: 各阶段为每项试验规定了三参数: #34、命中窗口、#34、#34、命中阈值、#34 和 #34; 初始阈值. 和 #34; 命中定义为在定义的时间段内达到标准角度。#34; 命中窗口和 #34; 指为动物开始试验和达到标准角度而分配的时间。#34; 命中阈值和 #34; 是成功试用的标准角度。#34; 初始阈值和 #34; 是启动试验和命中窗口的标准角度;这表示轮的开始。有关阶段的更多信息可以在下面的小节和 #34 中看到; 训练动物. 和 #34; </li> <li> 单击并 #34; 开始和 #34; 开始培训课程。点击和 #34; 饲料和 #34; 手动喂养动物和 #34; 停止和 #34; 停止培训会话. 
		注: 应使用手动喂养来保持对任务的兴趣, 详见3节。停止会话后, 会话数据将保存到 C: 驱动器. </li>
	</ol> </li> </ol> 2。生活和饲养条件 <ol> <li> 使用成年雌性大大鼠 (175-200 克)。将老鼠放在标准的笼子里, dark/12 12 h 的光周期。在黑暗的周期中执行所有的训练和测试, 灯光变暗. 
	注意: 雌性大鼠被使用, 因为它们更容易训练 21 。这里使用的是大, 因为它是一种常见的用于运动训练的菌株, 是转基因的主要菌株。但是, 考虑到任务的可调性质 (配重、转角标准、到旋钮的距离和旋钮本身), 这些可以很容易地调整为不同的菌株和更大的 ( 例如 雄性) 或更小的 ( 例如 大鼠). </li>
	<li> 在研究的前5天, 给老鼠一个完整的水槽, 除了营养完整, 巧克力味丸五天。第五天后, 食品限制大鼠的正常体重曲线的 85%. </li>
	<li> 食物限制开始后, 在整个训练过程中, 给老鼠喂食7.5 克巧克力丸 (调整上或下, 保持85% 的年龄调整体重)。如果老鼠在训练期间没有收到7.5 克巧克力丸, 在最后一次会议后补充 withstandard. 
	注: 周末喂食是 ad 随意 , 标准的餐食到星期日晚上, 然后食物就会在一夜之间被剥夺。当进行手术或其他侵入性干预, 允许食物 ad 随意 至少三天之前, 然后三天后的程序 (取决于恢复时间), 然后再把老鼠的食物限制. </li>
</ol> 3。培训过程 注意: 培训过程的概述见 图 3A 。在整个协议中, 每天训练两次老鼠, 一次在早晨, 一次在下午。在下午的会话开始之前至少要等待3小时. 
<ol> <li> 习惯 注意: 习惯的目标是让老鼠熟悉测试箱和处理。
	<ol> <li> 通过按自动定位器上的向下箭头键来完全收回旋钮设备. </li>
		<li> 将鼠标放在行为框中15分钟 </li>
		<li> 在箱中15分钟后, 将手里的每只老鼠至少五分钟, 使老鼠熟悉实验者. </li>
		<li> 重复此操作五天. </li>
	</ol> </li> <li> 奖励关联 注: 目标是训练鼠把旋钮的转动与食物奖励联系起来。期望在奖励协会阶段的培训期间, 花费30分钟的时间来完成整个会议。
	<ol> <li> 打开软件并输入 rat 名称。将舞台设置为和 #34; K1: 旋钮整形-没有滑轮. 和 #34; 注意: 这将 manipulandum 的距离设置为0.0 厘米、#34 和 Init 的光圈。打, #34; 3 #176; #34; 在5度, #34; 命中窗口和 #34; 在2秒。只要老鼠把旋钮转过3度, 老鼠就可以喂食。#34; 命中窗口和 #34; 将在整个培训过程中保持2秒. </li>
		<li> 把老鼠放在行为盒里, 按下 #34; 开始和 #34; 开始会话. </li>
		<li> 让老鼠熟悉奖励槽, 每次分配 2-3 粒, 然后敲击奖励槽所在的盒子的一侧。使用手动进纸器按钮分配小球. </li>
		<li> 一旦老鼠知道了奖赏的位置, 当老鼠在光圈前面时, 手动分配1小球. </li>
		<li> 一旦老鼠移动到光圈的预期的小球奖励, 条件鼠与 manipulandum 的互动。
		<ol> <li> 提示老鼠在 manipulandum 附近放置一个45毫克的食物颗粒, 或者直接将颗粒状的灰尘施加到 manipulandum, 让它伸出来抓住它。如果老鼠从光圈移开, 点击靠近 manipulandum 的盒子来重定向它的注意力. 
			注意: 如果老鼠用它的爪子触碰了旋钮, 程序会给老鼠喂食。任何不包括使用爪子的交互 (即咬、manipulandum) 都是不正确的, 不应该得到奖赏. </li>
		</ol> </li> <li> 一旦鼠开始接触所需的爪子的旋钮, 并获得奖励10连续时间在0.0 厘米, 使用向下箭头键的 auto-positioner ( 图 1A ), 以收回 manipulandum 0.25 厘米. 重复这在0.25 厘米增量, 直到 manipulandum 是 1.25 cm 离光圈. </li>
		<li> 按 #34; 停止和 #34; 在30分钟后结束会话. </li>
		<li> 继续奖励协会, 直到鼠 supinates manipulandum 与所需的爪子在1.25 厘米以外的光圈 ( 图 1B ). </li>
		<li> 一旦老鼠完成了2连续的疗程, supinating manipulandum 和回收小球奖励, 开始配重训练. </li></ol> </li> <li> 配重训练对老鼠进行训练, 以 supinate 6 克重的砝码. 
	<ol> <li> 在 manipulandum 上放置一个3克的配重, 通过将配线器连接到 manipulandum 上的 L 形附着点的一端。将配重柱通过滑轮送到平衡悬挂. </li>
		<li> 打开软件并输入鼠名。设置舞台和 #34; K2: 旋钮成型-滑轮. 和 #34; 注意: 这将 manipulandum 距离的光圈设置为1.25 厘米、#34 和 Init。打; #34; 3 #176; 和 #34; 打打 #34; 5 度。manipulandum 距离将保持在1.25 厘米从这一点向前. </li>
		<li> 把老鼠放在行为盒里, 按下 #34; 开始和 #34; 开始会话. </li>
		<li> 按 #34; 停止和 #34; 经过 100 + 成功的试验。一旦老鼠完成了连续2次疗程的 100 + 成功的试验, 增加重量1克后结束的会议。将重量从3克增加到6克, 在随后的会话中. </li>
		<li> 连续2次旋在 6 g 和 100 + 成功试验后继续执行步骤 3.4. 
		注: 平均需要6疗程 (3 天), 从3克到6克. 从配重训练开始, 正确的旋运动得到奖赏和不正确的旋运动的形成是势在必行的。有关正确和不正确的旋表单的可视化指南和说明, 请参阅 图 2 . </li>
	</ol> </li> <li> 对基线的训练 注意: 请记住继续塑造正确的旋运动。同样, 请参阅 图 2 , 以了解正确/不正确的旋移动的可视指南以及如何改善不正确的旋。训练大鼠 supinate 到基线标准;在这里, 这是75度与6克的平衡, 在75% 或更高的成功率。
	<ol> <li> 在 manipulandum 上放置一个6克重平衡. </li>
		<li> 打开软件并输入鼠名。设置舞台和 #34; KSB4: 旋钮训练中位数75最大和 #34; 注意: 这将设置和 #34; Init。打 #34; 5、#176; #34; 命中阈值最小和 #34; 在15和 #176;, #34; 命中阈值最大和 #34; 在75和 #176;。这就是所谓的 #34; 适应和 #34; 训练阶段, 意味着阈值将随着大鼠和 #39 的性能提高而增加。对于 #34; KSB4 和 #34; 阶段, 阈值首先设置为上一个会话和 #39; s 最终阈值。如果在此阶段没有运行前一个会话, 则阈值设置为最低15度的阈值。在前10次开庭试验后, 阈值被计算为先前10试验中峰值角的中点。因此, 每个试验的门槛是不同的, 取决于大鼠和 #39; 在以前的试验中的表现. </li>
		<li> 在30分钟后停止会话 </li>
		<li> 使用自适应阶段继续训练, 直到平均峰值角度为75度或更大。然后, 继续步骤3.5。这通常发生在大约10天或20次会话之后. </li>
	</ol> </li> <li> 基线评估记录四连续基线 <ol> <li> 在 manipulandum 上放置一个6克的平衡. </li>
		<li> 打开软件并输入鼠名。将舞台设置为和 #34; K27:75 度. #34; 这将设置和 #34; Init。打; #34; 5、#176; 和 #34; 命中阈值在75和 #176;. </li>
		<li> 按下 #34; 停止和 #34; 在30分钟或100次试验后, 无论是谁先来, 结束会话. </li>
		<li> 继续, 直到有四连续的基线, 成功率为75% 或更高. </li>
	</ol> </li> </ol> 4。损伤后评估 <ol> <li> 打开软件并输入鼠名。将舞台设置为与前基线测试相同的静态阶段。这将保留用于基线的相同参数. </li>
	<li> 按 #34; 启动和 #34; 运行会话直到30分钟 
	注意: 受伤后的参与程度可能较低。这可以通过使用手动送纸按钮或在 manipulandum 附近放置一个奖励颗粒来进行补救, 以诱使老鼠参与 manipulandum. </li>
	<li> 每星期重复测试一次, 直到达到所需的后评估长度. 
	注: 在这里, 我们评估后的表现每周六周. </li>
</ol> 5。分析数据 注意: 将数据保存到 PC 的 C: 驱动器上的默认位置。在命中窗口中, 数据以 100 Hz 的方式捕获, 并以二进制格式存储。在这里, 数据被分析使用一个定制的程序, 称为灵巧。请电子邮件 Dr. 杰森卡梅尔访问此免费软件. 
<ol> <li> 打开灵巧并单击和 #34; 标准. 和 #34; </li> <li> 查找 rat 数据所在的目录, 并选择所需的文件夹. </li>
	<li> 在选择目录后, 选择要分析的大鼠. </li>
	<li> 单击和 #34; #34 或 #34;D iscard 和 #34; 保留或丢弃不完整或不包含任何数据的文件. 
	注意: 有些时候, 会话是使用错误的参数和训练阶段开始的, 而当它们立即被停止时, 仍然会创建一个文件。此文件需要在分析期间丢弃。也有一些例子, 特别是在受伤后, 当一个动物不进行试验, 因为他们的伤害或很少的试验。对于这些会话, 必须保留一个文件, 因为它是其性能的表示形式。当没有成功试验的文件被保留时, NaN 被放置在适当的位置以计算变量. </li>
	<li> 选择是现在还是稍后对实验进行批注; 如果和 #34; 现在批注和 #34; 选定, 将打开一个新窗口。键入和 #34; 实验名称、#34; 和 #34; 组名称、#34; 和 #34; 事件数据、#34; 和 #34; 在实验中的总周数和 #34; 在试验中. </li>
	<li> 将主题分配给组. </li>
	<li> 输入每个星期或时间点的数据会话数。按空格分隔输入会话, 并确保在最后一个数字 即 (1 1 1 1) 之后不放置空格. </li>
	<li> 输入时间点标签。按空格分隔输入标签, 并确保在最后一个标签 即 (前 Wk1 Wk2 Wk3) 之后不放置空格. </li>
	<li> 选择事件发生的时间; 将显示保存分析会话的选项。单击并 #34; 是和 #34; 保存分析会话. </li>
	<li> 将显示注释和 unannotated 数据的概览。这提供了绘制数据的选项. </li>
	<li> 对于 unannotated 数据, 请单击和 #34; 图形主题和 #34; 绘制单个主题或 #34; 图表主题和 #34; 绘制所有主题在同一图表上. </li>
	<li> 对于注释分析, 请单击 "#34;P #34", 以绘制实验. </li>
	<li> 对于注释和 unannotated, 请单击要绘制的变量列表的下拉箭头。单击该变量以查看绘图. </li>
	<li> 单击 "导出" 以导出图形和/或与关系图关联的数据. </li>
</ol>

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Dr. Rennaker 和 Dr. 斯隆是 Vulintus Inc. 的所有者, 该公司在本出版物中制造设备。没有为其他作者声明利益冲突。

旋钮旋任务评估前肢旋在大鼠使用定量和半自动方法。为了实现这些端点, 为任务设计的许多参数, 包括旋钮对准、manipulandum 设计和训练标准, 都经过了数年的反复迭代。对于旋钮的对准, 我们试验了三不同的对齐旋钮与光圈: 左边的旋钮与左边的光圈, 以光圈为中心的旋钮, 和右侧的旋钮与右上侧对齐perture我们在右边的旋钮与光圈的右侧对齐, 因为这产生了在最短的时间内接受训练的大鼠, 并且后了最小的补偿机制, 特别是左爪的干扰。
至于 manipulandum 设计, 我们改变了几个设计特点, 以最大限度地利用前肢和尽量减少使用身体。此外, 我们将任务的难度扩大到预测赤字的严重性。pyramidotomy 后, 旋是最受影响的运动, 但损害仍然相对微妙。因此, 我们训练老鼠到一个更高的基线标准 (75), 以确保在受伤后观察到大量的赤字。对于更具损害性的皮质病变, 60 度阈值在 7.5 g 时足以证明损伤后的严重缺损。通过试验和错误方法优化的附加参数包括孔径大小、从光圈到旋钮的距离以及时间窗, 以获得成功的试验。
在整个培训协议中都有一些关键点需要仔细的监督。在对基线进行训练时, 采用自适应阈值法对大鼠进行 75 10训练。然而, 大鼠的高原峰值角度低于 75;在4-5 次会议之后, 性能仍然保持不变。为了提高性能, 可以使用静态阈值。静态阈值是指在设定的程度上保持的阈值, 它与大鼠的性能无关, 而不是根据最近的性能变化的自适应阈值。如果在适应训练中的大鼠高原, 实验者应改为静态阈值。静态训练阶段的范围从20到70度, 以10°递增。(舞台 K28-K33)。根据前2次会议中老鼠的平均峰值角度选择静态阶段。例如, 如果老鼠的平均45°, 选择静态阶段为50度 (K31)。所有静态阶段都设置 "Init"。打. "在5。在训练过程中, 如果老鼠失去动力, 如果它 supinates 接近但不超过阈值, 人工喂养老鼠。
此外, 在基线评估中, 大约5% 的大鼠在旋角和 5-10% 之间在两次会议之间的成功率上倒退 5-10 °。如果发生这种情况, 和鼠不恢复75平均峰值角后, 3-4 疗程, 减少静态阶段, 以10度的大鼠的当前平均角度之前返回步骤3.5。一旦把老鼠放在静态训练阶段, 就不要再把它重新引入适应阶段。
这项任务有一些限制。一旦建立了不正确的抓取位置, 改变抓取行为 (图 2) 可能会很困难。因此, 早期发现和纠正是很重要的。为了纠正老鼠的掌握, 光圈可以通过缩小水平和/或垂直方向的光圈大小来改变;通常我们将一张玻璃片贴在光圈边缘, 需要调整。对大多数老鼠来说, 这提高了他们的掌握形式, 因为它迫使他们以一种特定的方式抓住 manipulandum。这反过来又提高了他们正确 supinate 的能力。
除了这一挑战, 老鼠可以发展补偿机制, 以 supinate。这些包括使用头部, 以帮助前肢在旋;降低弯头和肩关节转动旋钮;使用左爪来帮助转动旋钮或推动到达的爪子向下。所有这些行为都可用于成功完成任务。如上所述, 与掌握有关的行为可以通过操纵光圈来纠正。补偿机制在掌握外面, 然而, 要求活跃参与由实验者不奖励报偿行为。在受伤后, 我们观察到老鼠在 supinating 前进行了几次试验, 把爪子放在合适的位置。虽然我们没有分析任务的哪些组成部分可能会导致旋的损失, 但这些可能包括失去准确的握持和削弱的力量调制, 在许多可能性中。
半自动旋任务平均需要20±5天的时间来训练老鼠的基线, 25% 的动物无法接受这项任务的训练。促进训练时间的事实是, 我们没有选择自然 right-preference 的老鼠, 而是迫使所有动物使用他们的右爪子, 是常见的最接近的化验。我们没有尝试使用左偏好的老鼠, 但这将是一个有趣的探索性研究, 首先确定爪偏好, 然后训练主导爪。为了适应这种情况, 我们需要翻转门的方向, 使光圈反转;这很容易做到。
与传统的 IBB 或单颗粒的任务相比, 旋任务定量、客观地衡量前肢的实现。它表现出对严重损伤 (皮质损害) 和细微损伤 (pyramidotomy) 的敏感性, 根据损伤模型的严重程度, 可以对训练程序进行修改。因为它是半自动, 任务允许实验者同时训练多只老鼠, 取决于训练阶段。这大大提高了实验者的生产率和老鼠的吞吐量。这项任务是可靠和可再生的老鼠之间。通过创建诊断指南 (图 2), 让实验者在培训协议中参考, 我们已经标准化了几个错误的行为以及解决方法。最后, 这项任务提供了一个直观的方法来分析大量的数据, 并使实验者能够深入研究旋的动力学。
将来, 我们将使用半自动旋任务作为评估康复的类型、剂量和时机的平台。我们的实验室对刺激对损伤后功能改善的影响感兴趣。此外, 我们感兴趣的是如何刺激神经修复或改善神经传导和沟通的疗法会影响康复。我们也有兴趣修改的任务, 以配合与电生理学, 使我们可以学习电机学习;大鼠头盖帽通常执行任务, 并且增加一个换向器为录音或刺激将是简单的做。这项任务, 如所述, 是为老鼠, 但也有实验室试验使用鼠标的任务。一般来说, 这项任务可用于评估前肢功能在啮齿动物的各种损伤模型和疾病状态, 反过来, 评估康复策略。前进, 我们将继续改进的任务, 通过改进, 以帮助减少错误的行为, 提高任务获取率和培训时间。

神经系统损伤或疾病后的灵巧性丧失显著降低受影响个人的独立性和生活质量1,2,3,4。因此, 灵巧性是理解神经修复和康复科学的重要结果措施, 也是运动和运动神经学习的神经控制的基础。传统上, 手工任务, 如单颗粒到达, 面食操作, 和欧文, Beatties, 和 Bresnahan (IBB) 前肢规模已被用来评估灵巧的动物, 特别是啮齿动物5, 6,7。这些任务由于其最短的任务获取时间而变得普及。然而, 他们是定性的在自然, 费力为实验者, 并且, 有时, 不敏感到功能损伤在损伤以后以微妙的缺乏5,7,8,9。这些传统任务的局限性促使了动物运动功能的定量测量的发展, 特别是前肢达到。
自动化任务有几个优点, 即客观性、增加的吞吐量和减少的分析时间。新的自动任务提供了比常规任务8,10更灵敏的评估损伤后的灵活性的措施。此外, 他们允许适应训练和测试, 裁缝训练和测试困难的动物的表现。最后, 自动化任务生成大量数据, 这提供了两个优点。首先, 在试验中和试验次数上增加数据增加了一项研究的统计能力。其次, 它为神经科学家提供了一个更大的数据集, 通过分析运动和运动学信息11来更有力地研究运动学习、训练和补偿。
几个组试图自动执行传统任务。高速摄像机可用于从任务中收集运动数据, 如单颗粒到达任务12。Alaverdashvili 和威肖使用高速摄影机捕捉到达的动作, 并使用帧-帧运动测量软件峰值茅图斯13来分析数字移动。但是, 该软件不使用计算机视觉识别数字, 而是要求实验者通过游标将移动点数字化。此外, 一些任务已与馈线和保持架一起使用, 以自动化培训过程14,15,16。
其他小组使用了力量传感器并且高速照相机评估空间调整和力量在熟练的前肢到达使用面食操作, 而其他组织设计了任务捕捉更加复杂的运动17。一个这样的任务是一个伸手可及的任务, 使用三自由度的机器人装置来捕捉大鼠前肢运动的平面和旋转运动18。这在能够测量运动的动能, 但随着复杂性和成本的增加而具有优势。
在这里, 我们演示了一个半自动的前肢任务, 即在大鼠8中测量旋。前肢旋是从手掌到手掌的旋转的爪子。旋是一个优秀的标志脊髓道功能和临床相关的运动在人类的日常活动所需的8,19,20。此外, 旋是高度敏感的伤害和失活, 特别是当与单颗粒达到8。旋任务, 在伯克医学研究所和德克萨斯大学达拉斯分校的合作下, 在水平平面8,10中测量旋转运动。鼠被放置在一个行为盒 (图 1A) 和训练做三运动 (图 1B): 通过一个长方形的光圈;掌握球形 manipulandum;supinate 到指定的角度。
行为任务由 PC 软件 (图 1C) 控制。控制软件向连接到 auto-positioner、光编码器、扬声器和馈线的微控制器发送指令。微控制器及其外围连接称为微控制器盒。信息流从光编码器, 到单片机, 再到电脑, 再回到微控制器。如果控制软件已经向微控制器发出信号, 证明试验是成功的, 微控制器会触发送纸器来分配小球。在每个会话开始时, 控制软件将舞台信息中继到单片机, 它指示 auto-positioner 将旋钮放置在舞台上的指定距离的光圈。auto-positioner 还可以使用位于 auto-positioner 上的箭头键手动操作。光编码器以100赫兹的方式记录数据, 并测量角度的变化。所有数据都以二进制格式存储。
实验者使用软件中的顺序训练阶段, 以预定的角度和成功率训练大鼠从适应到 supinating。在习惯, 旋钮 manipulandum 是放置在光圈窗口没有任何平衡。经过一周的高度监督训练, 老鼠把旋钮与奖励联系起来, 开始独立转动旋钮。一旦老鼠能够独立地转动, 旋钮被缩回到 1.25 cm 在 0.25 cm 增量, 直到老鼠能独立地转动在 1.25 cm. 然后, 配重从 3 g 到 6 g 增加 1 g. 自动培训阶段训练动物 supinate 旋钮在6克到75度这一阶段的培训基本上是无监督的;一旦老鼠采取适当的形式 (下面讨论) 的任务, 他们继续 supinate 正确。训练完成时, 老鼠 supinate 75 度的成功率 (命中率) 为 75% 8。在这里, 我们描述了一个典型的培训协议, 并针对我们遇到的常见问题提出了解决方案。我们通过训练协议来证明有代表性的成功和不成功的老鼠的进展, 并表明这项任务可以被修改, 以显示功能性损伤与微妙或更严重的赤字。

<table><tbody><tr><td>Base Cage - Rat Model</td><td>Vulintus</td><td>MotoTrak Rat System</td><td>N/A</td></tr><tr><td>Controller</td><td>Vulintus</td><td>MotoTrak Rat System</td><td>N/A</td></tr><tr><td>Behavior Module</td><td>Vulintus</td><td>MotoTrak Rat System</td><td>Supination Task, Methacrylate Dual Stop Knobs</td></tr><tr><td>Pellet Dispenser - 45mg</td><td>Vulintus</td><td>MotoTrak Rat System</td><td>N/A</td></tr><tr><td>Autopositioner</td><td>Vulintus</td><td>MotoTrak Rat System</td><td>N/A</td></tr><tr><td>45 mg, Chocolate Flavor, 50,000/Box</td><td>Bio-Serv</td><td>F0299</td><td>N/A</td></tr><tr><td>HP Z230 Tower WorkStation</td><td>HP</td><td>N/A</td><td>Intel Xeon CPU E3-1225 v3 @ 3.20 GHz, 16GB RAM, 1TB HDD. Min Requirements: 8GB RAM, Multi-Core Processor</td></tr><tr><td>Dexterity</td><td>Burke Medical Research Institute</td><td></td><td>Matlab software for data analysis</td></tr><tr><td>Enviropak</td><td>WF Fisher and Son</td><td>N/A</td><td>N/A</td></tr></tbody></table>

the knob supination task: a semi-automated method for assessing forelimb function in rats

准确测量动物模型灵巧度的任务是理解手功能的关键。目前, 测量灵巧度的大鼠行为任务主要使用对接触或食物操作的视频分析。虽然这些任务很容易实施, 并且在疾病模型中很有活力, 但对实验者来说却是主观和费力的。自动化传统任务或创建新的自动化任务可以使任务更加高效、客观和定量。由于老鼠的灵巧性不如灵长类动物, 中枢神经系统 (CNS) 的损伤在灵巧度上会产生更微妙的缺陷, 然而, 旋在啮齿类动物中受到高度影响, 对灵长类的手功能至关重要。因此, 我们设计了一个半自动的任务, 测量前肢旋在大鼠。老鼠被训练到达并且掌握一个旋钮形的 manipulandum 并且转动 manipulandum 在旋接受报偿。老鼠可以在20±5天内获得这项技能。虽然培训的早期部分受到高度监督, 但大部分培训是在没有直接监督的情况下进行的。该任务可靠和性捕捉微妙的缺陷后受伤, 并显示功能恢复, 准确反映临床恢复曲线。对数据的分析是由专门的软件通过图形用户界面来完成的, 设计的直观。我们还给出了在训练过程中遇到的常见问题的解决方法, 并表明在训练初期对行为的轻微纠正会产生可靠的旋。因此, 旋钮旋任务提供了有效和定量评价的关键运动的灵巧在大鼠。

在训练的早期, 实验者花更多的时间在任务上塑造老鼠的行为。当老鼠把老鼠旋与奖赏联系在一起时, 手上的时间就会减少 (图 3A)。在习惯, 奖励协会, 和配重训练中, 整个会话长度 (30 分钟) 用于任务。然而, 当老鼠 supinating 6 克体重后, 当老鼠的旋角增加时, 任务的时间逐渐减少到约15分钟。最后, 当鼠达到基线时, 任务时间最短;实验者只需要把老鼠放在行为箱里, 然后启动程序。实验者可以同时工作的老鼠的最大数量是两只老鼠在奖励协会期间, 四只老鼠在配重训练和训练到基线, 和许多老鼠一样有箱子在基准评估和后测试期间。平均来说, 75% 的老鼠 (n = 56) 获得这项任务。
在老鼠与奖励相关的旋后, 在老鼠的旋角 (图 3B) 中有一个积极的进展。在图 3B中, 从3天到7天, 大鼠从3克到6克的平衡进展。在配重训练后, 从7天到9天的自适应训练的时间较短, 其间旋从26到30度增加。由于没有太大的变化, 静态阈值从9天到18使用。在此期间, 老鼠在8天内稳步从30度增加到75度。在整个培训中, 特别是在12和14天, 每天都有变化。但总的来说, 旋的角度有上升的趋势。在17天的 post-habituation, 老鼠已经记录了它的第一个基线, 四次会议后, 它完成了基线评估。从适应到记录第四基线, 训练协议平均需要20±5天。
虽然通过培训协议查看理想的进程非常重要, 但查看不成功的进程同样重要 (图 4)。在图 4A中, 橙色线显示成功完成协议的 rat, 蓝线显示不成功的鼠, 灰线显示另六成功的老鼠。成功的老鼠在15±0.6 天到达了基线 (n = 7)。代表成功的老鼠用1点的时间掌握, 而不成功的老鼠使用3点的时钟掌握。两只老鼠把这个旋钮与2天内的奖赏联系起来。此外, 两个大鼠显示一个类似的旋角 (图 4A) 的进展, 在前四天的配重增加。然而, 在这一点之后, 成功的老鼠开始摆脱不成功的老鼠。这是因为不成功的老鼠的掌握无法在这一点之前得到纠正 (参见图 2)。
对于成功的老鼠, 有一个陡峭的上升旋角, 开始在50和60度之间的高原, 但随后恢复到75度的稳步攀升。然而, 对于不成功的老鼠, 有一个更逐步增加的旋角。当老鼠高原大约20度时, 老鼠被推挤到 supinate 更多, 但最终, 它失去兴趣在任务, 甚而以手工哺养, 并且旋角度迅速减少在天附近 15 post-habituation。虽然在17天 post-habituation 后有轻微的恢复, 但老鼠挣扎着 supinate 超过25度。如果老鼠没有达到20天的基线, 我们认为这只老鼠不成功, 并从研究中移除老鼠。
除了旋角度外, 还可以对成功和不成功的老鼠进行旋波形 (图 4B-d) 的视觉检查。在执行视觉检查时, 我们会查找几个波形特征: 线的斜率、延迟和试验时间窗口中的峰值数。该线的斜率计算为曲线起始曲线与曲线峰值之间的导数。延迟的计算方法是在开始试验和越过命中阈值的曲线之间的时间。最后, 利用导数在试验窗口中求出极值, 计算出峰值。以前, 我们发现, 线的斜率, 或速度, 是一个强大的测量旋动力学和敏感的微妙赤字8。
在训练的前三分之一开始到 supinate 使用 6 g (图 4B), 成功的鼠 (图 4B1) 显示一个波形, 峰值接近20度, 而不成功的鼠 (图 4B2) 显示双转, 或两个峰值, 第一峰值接近10度, 第二峰值接近5度。在训练的中间三分之一 (图 4C) 中, 成功的大鼠 (图 4C1) 显示峰值角度从20度增加到50度, 并具有更明确的单峰曲线。不成功的老鼠 (图 4C2), 同时, 只显示在峰值角度的边际增加到20度, 但改善了它的形式;它现在只使用一个单一的转弯。在训练的最后三分之一 (图 4D) 中, 成功的大鼠 (图 4D1) 显示了一个非常明显的单波形, 峰值围绕 65, 而不成功的鼠 (图 4D2) 的峰值角为20度, 但现在有一个额外的峰值在2秒的15。这是另一个很好的指标, 随着训练难度的增加, 大鼠无法纠正其3点的掌握, 反过来又无法正确 supinate。即使这只老鼠没有被排除在研究范围之内, 并且最终可能达到75度, 但问题仍然是关于它是否是真正的旋与旋的补偿。
最后, 旋任务检测多种类型的损伤后的功能损伤, 包括脊髓道的切口病变, 人的自愿运动的主要通路, 和前肢运动皮层病变的内皮素注射液(图 5) 8,10,22。大鼠在 pyramidotomy 组 (紫色, n = 8) 被训练到 supinate 至少75在 6 g 的成功率为75% 或以上, 而鼠皮层病变组 (绿色, n = 10) 被训练到 supinate 60 °7.5 或以上75% 克。两组大鼠在受伤后成功率明显下降 (图 5A)。pyramidotomy 组大鼠的成功率从90% ±2% 降至14% ±8%。成功皮质病变大鼠从76% ±1% 降至10% ±3%。到6周, 两组仍然受损: pyramidotomy 组是在34% ± 11%, 而皮质病变组保持在16% ±7%。对于旋角, 两个组显示售后 (图 5B) 的减少。由于不同的标准基线旋角, pyramidotomy 组有一个更高的前旋角 (85/85 ± 2.9°) 比皮层病变组 (67 ± 0.52°)。pyramidotomy 组减少至38±10°, 皮层病变组减少至27±2.9°。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56341/56341fig1.jpg"> 
图 1:旋任务描述(A) 老鼠被放在一个有机玻璃盒子里, 它有一个光圈, 通过它达到并抓住一个旋钮, 必须在旋中转动。旋钮有两个停止, 以防止旋角大于100。该旋钮还有一个滑轮与平衡;这创造了老鼠必须克服的扭矩 supinate。旋钮连接到一个光学编码器, 测量角度与0.25°的准确性。该光学编码器连接到单片机, 它反过来连接到控制任务的计算机。当一个成功的标准实现时, 计算机向单片机发出信号, 当触发音频反馈并从馈线中分配一个小球。该单片机还控制的位置在0和 1.25 cm 是由计算机设置的训练阶段口授的自动定位器。(B) 老鼠在三次连续的运动中执行任务: 通过光圈到达, 在1点的时候抓住旋钮, 并 supinating。(C) 旋钮旋任务由控制软件控制。实验者输入主体名称, 选择训练阶段, 程序设置相应的参数。一个成功的旋试验的波形以蓝色显示, 而成功和失败的试验顺序分别显示为绿色和红色。如果旋角度大于定义的时间窗口中的命中阈值, 则测试将被控制软件标记为成功, 而如果不这样做, 则试验将被标记为失败。此程序控制一个框。每台计算机可以同时运行四程序。此图已从 Sindhurakar et al.、2017、康复和神经修复8中进行了修改。<a href="//ecsource.jove.com/files/ftp_upload/56341/56341fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56341/56341fig2.jpg"> 
图 2:旋移动.在训练协议中遇到的常见正确和不正确的旋动作的图示和描述。正确的运动允许真正的旋, 而不正确的运动包括补偿机制, 可能会阻止真正的旋。包括建议的解决方案, 以纠正不正确的运动。<a href="//ecsource.jove.com/files/ftp_upload/56341/56341fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/56341/56341fig3.jpg"> 
图 3: 培训协议.(A) 标准时间线。总共有五期的培训持续约25天: 习惯性 (5 d)、奖励协会 (1-3 d)、举重训练 (3-4 d)、基线培训 (8-12 d) 和基线评估 (2-4 d)。时间线上的线条图案指定了实验者在每届会议上花费在任务上的时间。随着培训协议的进行, 任务时间减少。(B) 大鼠从奖赏协会 supinate 到基线评估的能力的一般进展。总的来说, 老鼠对基线有一个积极的线性进展, 但是正如观察到的, 在整个训练协议中, 老鼠的表现是有变化的。在举重训练后, 有一个适应训练的时期, 旋角阈值改变以符合大鼠的表现。这种自适应训练之后是一个静态的阈值模式, 直到老鼠已经达到基线。<a href="//ecsource.jove.com/files/ftp_upload/56341/56341fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/56341/56341fig4.jpg"> 
图 4:成功和不成功的任务获取(A) 旋角的进展整个训练协议为八只老鼠, 七成功和一个不成功。一个代表性的老鼠, 达到基线标准 (成功, 橙色) 和一个不成功的老鼠 (蓝色) 进一步被用作个案研究。在训练后的前七天中, 成功的和不成功的大鼠在旋角上表现出相似的进展。在11天 post-habituation, 成功的老鼠是 supinating 55, 而不成功的鼠后°。在15天的 post-habituation, 成功的老鼠表现出了强劲的上升, 而不成功的老鼠在性能下降。在训练 post-habituation 的最后三分之一, 不成功的老鼠平稳在30°, 而成功的老鼠是 supinating 80。(B) 平均波形 (黑线) 以95% 的置信区间 (橙色为成功, 蓝色为不成功) 的训练后三分之一的6克的平衡增加。(B1)成功的鼠-单峰周围°。(B2)不成功的鼠-双峰值与全球最大10°。(C) 平均波形 (黑线) 以95% 的置信区间 (橙色为成功, 蓝色为不成功) 的第二个训练后三分之一的配重的6克增加。(C1)成功的大鼠单峰在45°。(C2)不成功的老鼠改进的形式与单峰近°。(D) 平均波形 (黑线) 以95% 的置信区间 (橙色为成功, 蓝色为不成功), 为最后三分之一的训练后, 6 克的配重增加。(D1)成功的老鼠发音单峰在65。(D2)不成功的鼠-双峰值与全球最大°。<a href="//ecsource.jove.com/files/ftp_upload/56341/56341fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/56341/56341fig5.jpg"> 
图 5:较强的任务敏感性不同的伤害模型。大鼠在 pyramidotomy 组 (紫色, n = 8) 被训练到 supinate 75 在 6 g 的成功率为75% 或以上, 而大鼠皮层病变组 (绿色, n = 10) 被训练到 supinate 60 °6或以上75% 克。数据显示的是平均±标准误差。(A) pyramidotomy 病变与皮质病变的成功率。两个损伤模型显示售后 (1 周) 的成功率急剧下降。pyramidotomy 成功率从0.90 ±0.02 降至0.14 ± 0.08, 而皮质病变成功率从0.76 ±0.01 下降。(B) pyramidotomy 与皮质病变的旋角度。两组均显示售后: pyramidotomy 组减少至38.2°± 10.1°, 皮层病变组减少至27.1°±2.9°。<a href="//ecsource.jove.com/files/ftp_upload/56341/56341fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about The Knob Supination Task: A Semi-automated Method for Assessing Forelimb Function in Rats at JoVE.com

The Knob Supination Task: A Semi-automated Method for Assessing Forelimb Function in Rats

旋钮旋任务: 一种半自动评估大鼠前肢功能的方法

这份手稿描述了一个半自动的任务, 量化旋在大鼠。老鼠到达, 抓住, 并且 supinate 球形 manipulandum。如果转弯角度超过用户设定的标准, 老鼠就会得到一个小球的奖赏。与传统任务相比, 此任务增加了吞吐量、对伤害的敏感度和客观性。

Tasks that accurately measure dexterity in animal models are critical to understand hand function. Current rat behavioral tasks that measure dexterity ...

the-knob-supination-task-semi-automated-method-for-assessing-forelimb

Research

JoVE Journal

Behavior

9.2K 次观看.  Burke Medical Research Institute. 这份手稿描述了一个半自动的任务, 量化旋在大鼠。老鼠到达, 抓住, 并且 supinate 球形 manipulandum。如果转弯角度超过用户设定的标准, 老鼠就会得到一个小球的奖赏。与传统任务相比, 此任务增加了吞吐量、对伤害的敏感度和客观性。

视频: The Knob Supination Task: A Semi-automated Method for Assessing Forelimb Function in Rats

Assessing Forelimb Function after Unilateral Cervical SCI using Novel Tasks: Limb Step-alternation, Postural Instability and Pasta Handling

Cervical spinal cord injury (cSCI) can cause devastating neurological deficits, including impairment or loss of upper limb and hand function. A majority of the spinal cord injuries in humans occur at the cervical levels. Therefore, developing cervical injury models and developing relevant and sensitive behavioral tests is of great importance. Here we describe the use of a newly developed forelimb step-alternation test after cervical spinal cord injury in rats. In addition, we describe two behavioral tests that have not been used after spinal cord injury: a postural instability test (PIT), and a pasta-handling test. All three behavioral tests are highly sensitive to injury and are easy to use. Therefore, we feel that these behavioral tests can be instrumental in investigating therapeutic strategies after cSCI.

Three new behavioral tests (forelimb step-alternation, postural instability test, pasta handling test) for evaluating forelimb function after cervical spinal cord injury in rodents are described.

采用新型任务评估前肢功能单侧颈脊髓损伤后：肢体步交替，姿势不稳和面食处理

Cervical spinal cord injury (cSCI) can cause devastating neurological deficits, including impairment or loss of upper limb and hand function. A majority ...

科研

行为学

The Vermicelli and Capellini Handling Tests: Simple quantitative measures of dexterous forepaw function in rats and mice

Previous characterizations of rodent eating behavior have revealed that they use coordinated forepaw movements to manipulate food pieces. We have extended upon this work to develop a simple quantitative measure of forepaw dexterity that is sensitive to lateralized impairments and age-dependent changes. Rodents learn skillful forepaw and digit movements to manage thin pasta pieces, which they eagerly consume. We have previously described methods for quantifying vermicelli handling in rats and showed that the measures are very sensitive to forelimb impairments resulting from unilateral ischemic lesions, middle cerebral artery occlusions and unilateral striatal dopamine depletion [Allred, R.P., Adkins, D.L., Woodlee, M.T., Husbands, L.C., Maldonado M.A., Kane, J.R., Schallert, T. &amp; Jones, T.A. The Vermicelli Handling Test: a simple quantitative measure of dexterous forepaw function in rats. J. Neurosci. Methods 170, 229-244 (2008)]. Here we present a more detailed protocol for this test in rats and compare it with a newly developed version for mice, the Capellini Handling Test. Rats and mice are videotaped while handling short lengths of uncooked vermicelli or capellini pasta, respectively, with a camera positioned to optimize the view of paw movements. Slow motion video playback allows for the identification of forepaw adjustments, defined as any distinct removal and replacement of the paw, or of any number of digits, on the pasta piece after eating commences. Forepaw adjustments per piece are averaged over trials per each testing session. Repeated testing permits sensitive quantitative analysis of changes in forepaw dexterity over time. Protocols for pre-testing habituation and handling practice, as well as procedures for characterizing atypical handling patterns, are described. Because rats and mice perform the pasta handling tests slightly differently, species-specific differences in administration and scoring of these tests are highlighted. All animal use was in accordance with protocols approved by the University of Texas at Austin Animal Care and Use Committee.

The Vermicelli and Capellini Handling Tests of forepaw dexterity take advantage of the natural inclination of rodents to manipulate food items using skillful forepaw and digit movements. Animals are videotaped while handling short strands of uncooked dry pasta. Slow motion video playback allows for the quantification of forepaw adjustments.

粉条和Capellini处理测试：在大鼠和小鼠的灵巧的前爪功能简单的定量措施

Previous characterizations of rodent eating behavior have revealed that they use coordinated forepaw movements to manipulate food pieces. We have extended ...

神经科学

A Novel Method for Assessing Proximal and Distal Forelimb Function in the Rat: the Irvine, Beatties and Bresnahan (IBB) Forelimb Scale

Several experimental models of cervical spinal cord injury (SCI) have been developed recently to assess the consequences of damage to this level of the spinal cord (Pearse et al., 2005, Gensel et al., 2006, Anderson et al., 2009), as the majority of human SCI occur here (Young, 2010; www.sci-info-pages.com). Behavioral deficits include loss of forelimb function due to damage to the white matter affecting both descending motor and ascending sensory systems, and to the gray matter containing the segmental circuitry for processing sensory input and motor output for the forelimb. Additionally, a key priority for human patients with cervical SCI is restoration of hand/arm function (Anderson, 2004). Thus, outcome measures that assess both proximal and distal forelimb function are needed. Although there are several behavioral assays that are sensitive to different aspects of forelimb recovery in experimental models of cervical SCI (Girgis et al., 2007, Gensel et al., 2006, Ballerman et al., 2001, Metz and Whishaw, 2000, Bertelli and Mira, 1993, Montoya et al., 1991, Whishaw and Pellis, 1990), few techniques provide detailed information on the recovery of fine motor control and digit movement.
The current measurement technique, the Irvine, Beatties and Bresnahan forelimb scale (IBB), can detect recovery of both proximal and distal forelimb function including digit movements during a naturally occurring behavior that does not require extensive training or deprivation to enhance motivation. The IBB was generated by observing recovery after a unilateral C6 SCI, and involves video recording of animals eating two differently shaped cereals (spherical and doughnut) of a consistent size. These videos were then used to assess features of forelimb use, such as joint position, object support, digit movement and grasping technique.
The IBB, like other forelimb behavioral tasks, shows a consistent pattern of recovery that is sensitive to injury severity. Furthermore, the IBB scale could be used to assess recovery following other types of injury that impact normal forelimb function.

A Novel Method for Assessing Proximal and Distal Forelimb Function in the Rat: the Irvine, Beatties and Bresnahan IBB Forelimb Scale

Here we will describe a rodent behavioral assay that can detect recovery of both proximal and distal forelimb function including digit movements during a naturally occurring behavior that does not require extensive training or deprivation to enhance motivation.

一个评估在大鼠近端和远端的前肢功能的新方法：欧文，Beatties和布雷斯纳汉（IBB）前肢规模

Several experimental models of cervical spinal cord injury (SCI) have been developed recently to assess the consequences of damage to this level of the ...

Acquisition of a High-precision Skilled Forelimb Reaching Task in Rats

Movements are the main measurable output of central nervous system function. Developing behavioral paradigms that allow detailed analysis of motor learning and execution is of critical importance in order to understand the principles and processes that underlie motor function. Here we present a paradigm to study movement acquisition within a daily session of training (within-session) representing the fast learning component and primary acquisition as well as skilled motor learning over several training sessions (between-session) representing the slow learning component and consolidation of the learned task. This behavioral paradigm increases the degree of difficulty and complexity of the motor skill task due to two features: First, the animal realigns its body prior to each pellet retrieval forcing renewed orientation and preventing movement execution from the same angle. Second, pellets are grasped from a vertical post that matches the diameter of the pellet and is placed in front of the cage. This requires a precise grasp for successful pellet retrieval and thus prevents simple pulling of the pellet towards the animal. In combination with novel genetics, imaging and electrophysiological technologies, this behavioral method will aid to understand the morphological, anatomical and molecular underpinnings of motor learning and memory.

A paradigm is presented to analyze the acquisition of a high-precision skilled forelimb reaching task in rats.

收购高精度熟练的前肢到达任务大鼠

Movements are the main measurable output of central nervous system function. Developing behavioral paradigms that allow detailed analysis of motor ...

Investigating Motor Skill Learning Processes with a Robotic Manipulandum

Skilled reaching tasks are commonly used in studies of motor skill learning and motor function under healthy and pathological conditions, but can be time-intensive and ambiguous to quantify beyond simple success rates. Here, we describe the training procedure for reach-and-pull tasks with ETH Pattus, a robotic platform for automated forelimb reaching training that records pulling and hand rotation movements in rats. Kinematic quantification of the performed pulling attempts reveals the presence of distinct temporal profiles of movement parameters such as pulling velocity, spatial variability of the pulling trajectory, deviation from midline, as well as pulling success. We show how minor adjustments in the training paradigm result in alterations in these parameters, revealing their relation to task difficulty, general motor function or skilled task execution. Combined with electrophysiological, pharmacological and optogenetic techniques, this paradigm can be used to explore the mechanisms underlying motor learning and memory formation, as well as loss and recovery of function (e.g. after stroke).

A paradigm is presented for training and analysis of an automated skilled reaching task in rats. Analysis of pulling attempts reveals distinct subprocesses of motor learning.

调查运动技能的学习过程与机器人Manipulandum

Skilled reaching tasks are commonly used in studies of motor skill learning and motor function under healthy and pathological conditions, but can be ...

Behavioral Assessment of Manual Dexterity in Non-Human Primates

The corticospinal (CS) tract is the anatomical support of the exquisite motor ability to skillfully manipulate small objects, a prerogative mainly of primates1. In case of lesion affecting the CS projection system at its origin (lesion of motor cortical areas) or along its trajectory (cervical cord lesion), there is a dramatic loss of manual dexterity (hand paralysis), as seen in some tetraplegic or hemiplegic patients. Although there is some spontaneous functional recovery after such lesion, it remains very limited in the adult. Various therapeutic strategies are presently proposed (e.g. cell therapy, neutralization of inhibitory axonal growth molecules, application of growth factors, etc), which are mostly developed in rodents. However, before clinical application, it is often recommended to test the feasibility, efficacy, and security of the treatment in non-human primates. This is especially true when the goal is to restore manual dexterity after a lesion of the central nervous system, as the organization of the motor system of rodents is different from that of primates1,2. Macaque monkeys are illustrated here as a suitable behavioral model to quantify manual dexterity in primates, to reflect the deficits resulting from lesion of the motor cortex or cervical cord for instance, measure the extent of spontaneous functional recovery and, when a treatment is applied, evaluate how much it can enhance the functional recovery.
The behavioral assessment of manual dexterity is based on four distinct, complementary, reach and grasp manual tasks (use of precision grip to grasp pellets), requiring an initial training of adult macaque monkeys. The preparation of the animals is demonstrated, as well as the positioning with respect to the behavioral set-up. The performance of a typical monkey is illustrated for each task. The collection and analysis of relevant parameters reflecting precise hand manipulation, as well as the control of force, are explained and demonstrated with representative results. These data are placed then in a broader context, showing how the behavioral data can be exploited to investigate the impact of a spinal cord lesion or of a lesion of the motor cortex and to what extent a treatment may enhance the spontaneous functional recovery, by comparing different groups of monkeys (treated versus sham treated for instance). Advantages and limitations of the behavioral tests are discussed. The present behavioral approach is in line with previous reports emphasizing the pertinence of the non-human primate model in the context of nervous system diseases2,3.

As manual dexterity is a prerogative mainly of primates, behavioral tasks have been developed in macaque monkeys. Four reach and grasp prehension tasks, measuring hand manipulation ability and force, allow to establish functional recovery after a lesion of the central nervous system and to test the effect of a treatment.

在非人类灵长目动物行为评估手巧

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

Evaluating Skilled Prehension in Mice Using an Auto-Trainer

We describe a method to introduce na&#239;ve mice to a novel prehension (reach-to-grasp) task. Mice are housed singly in cages with a frontal slot that permits the mouse to reach out of its cage and retrieve food pellets. Minimal food restriction is employed to encourage the mice to perform the food retrieval from the slot. As the mice begin to associate coming to the slot for food, the pellets are manually pulled away to stimulate extension and pronation of their paw to grasp and retrieve the pellet through the frontal slot. When the mice begin to reach for the pellets as they arrive at the slot, the behavioral assay can be performed by measuring the rate at which they successfully grasp and retrieve the desired pellet. They are then introduced to an auto-trainer that automates both the process of providing food pellets for the mouse to grasp, and the recording of successful and failed reaching and grasping attempts. This allows for the collection of reaching data for multiple mice with minimal effort, to be used in experimental analysis as appropriate.

Method to assess the impact of training on motor skills is a useful tool. Unfortunately, most behavioral assessments can be labor intensive and/or expensive.We describe here a robotic method of assessing prehension (reach-to-grasp) skill in mice.

使用自动训练器评估小鼠的熟练前向

We describe a method to introduce na&#239;ve mice to a novel prehension (reach-to-grasp) task. Mice are housed singly in cages with a frontal slot that ...

Automated Rat Single-Pellet Reaching with 3-Dimensional Reconstruction of Paw and Digit Trajectories

Rodent skilled reaching is commonly used to study dexterous skills, but requires significant time and effort to implement the task and analyze the behavior. Several automated versions of skilled reaching have been developed recently. Here, we describe a version that automatically presents pellets to rats while recording high-definition video from multiple angles at high frame rates (300 fps). The paw and individual digits are tracked with DeepLabCut, a machine learning algorithm for markerless pose estimation. This system can also be synchronized with physiological recordings, or be used to trigger physiologic interventions (e.g., electrical or optical stimulation).

Rodent skilled reaching is commonly used to study dexterous skills, but requires significant time and effort to implement the task and analyze the behavior. We describe an automated version of skilled reaching with motion tracking and three-dimensional reconstruction of reach trajectories.

自动大鼠单丸到达与3维重建的爪子和数字轨迹

Rodent skilled reaching is commonly used to study dexterous skills, but requires significant time and effort to implement the task and analyze the ...

Unilateral Pyramidotomy of the Corticospinal Tract in Rats for Assessment of Neuroplasticity-inducing Therapies

The corticospinal tract (CST) can be completely severed unilaterally in the medullary pyramids of the rodent brainstem. The CST is a motor tract that has great importance for distal muscle control in humans and, to a lesser extent, in rodents. A unilateral cut of one pyramid results in loss of CST innervation of the spinal cord mainly on the contralateral side of the spinal cord leading to transient motor disability in the forelimbs and sustained loss of dexterity. Ipsilateral projections of the corticospinal tract are minor. We have refined our surgical method to increase the chances of lesion completeness. We describe postsurgical care. Deficits on the Montoya staircase pellet reaching test and the horizontal ladder test shown here are detected up to 8 weeks postinjury. Deficits on the cylinder rearing test are only detected transiently. Therefore, the cylinder test may only be suitable for detection of short term recovery. We show how, electrophysiologically and anatomically, one may assess lesions and plastic changes. We also describe how to analyse fibers from the uninjured CST sprouting across the midline into the deprived areas. It is challenging to obtain &#62;90% complete lesions consistently due to the proximity to the basilar artery in the medulla oblongata and survival rates can be low. Alternative surgical approaches and behavioural testing are described in this protocol. The pyramidotomy model is a good tool for assessing neuroplasticity-inducing treatments, which increase sprouting of intact fibers after injury.

The corticospinal tract, one of the major sensorimotor tracts, can be lesioned unilaterally in the rodent brainstem in order to test neuroplasticity-inducing therapies for the central nervous system. This surgical procedure (&#8220;pyramidotomy&#8221;) and postoperative assessments are described in this protocol.

皮质脊髓束的大鼠单侧锥体束的神经可塑性诱导疗法的评估

The corticospinal tract (CST) can be completely severed unilaterally in the medullary pyramids of the rodent brainstem. The CST is a motor tract that has ...

Massive Pontine Hemorrhage by Dual Injection of Autologous Blood

We provide a protocol to establish a massive pontine hemorrhage model in a rat. Rats weighing about 250 grams were used in this study. One hundred microliters of autologous blood was taken from the tail vein and stereotaxically injected into the pons. The injection process was divided into 2 steps: First, 10 &#181;L of blood was injected into a specific location, anteroposterior position (AP) -9.0 mm; lateral (Lat) 0 mm; vertical (Vert) -9.2 mm, followed by a second injection of the residual blood located at AP -9.0 mm; Lat 0 mm; Vert -9.0 mm with a 20-minute interval. The balance beam test, limb placement test, and the modified Voestch neuroscore were used to evaluate neurological function. Magnetic Resonance Imaging (MRI) was used to assess the volume of hemorrhage in vivo. The symptoms of this model were in line with patients with massive pontine hemorrhage.

We present a protocol to establish a massive pontine hemorrhage model in a rat via dual injection of autologous blood.