本出版物是通过固化CMD，法国肌肉营养不良症协会（AFM），从肌肉萎缩症协会，健康（1K26RR032082，1P50AR060836-01，1U54HD071601，2R24HD050846-06）全国学院，国防部平移研究资助（资助W81XWH，11-1-0330，W81XWH，11-1-0782，W81XWH，10-1-0659，W81XWH，11-1-0809，W81XWH-09-1-0599）和父项目肌肉营养不良症的试点补助（ PPMD）。 本文是几个在一系列的SOP在先天性肌肉疾病领域经常使用的方法之一。它反映了努力，在最近的先天性肌肉疾病协会研讨会，2013年4月在华盛顿特区举行的讨论，并成立了由20名专家先天性肌肉疾病领域

注：开放现场活动监控系统采用开放式实地有机玻璃室与光电发射器和受体同样沿室的周边间隔（ 图1）。这些光电发射器和受体产生不可见的红外光束的XY网格中。当动物被放置在腔室中，它的动作有关，从而导致光束中断。垂直传感器也存在评估垂直活动水平（ 即饲养行为）也是如此。该分析仪记录了梁断裂的信息，并迅速对其进行分析。的计算机软件，然后计算在预定的时间周期的多个活动的措施。这些措施包括：水平的活性（单位），垂直活动（单位），总行驶距离（厘米），动作时间（秒），和其余的时间（秒）5。 注：在一般考场应该是温度和湿度的控制，具有均匀的照明。试验室应均匀存款保险计划祭牲在房间里，不应该放在直射光线，黑暗的角落，或阴影区域。所有仪器驯化和测试应在每天的同一时间执行（ 例如 ，在上午）和由相同的个人。这些人应该被蒙蔽的动物治疗组和基因型在可能的情况。 下面的协议已指导和全国儿童医疗中心IACUC批准下被执行。 1，仪器驯化<ol><li>将小鼠在试验室在他们的家笼约10分钟，以适应新环境。留在驯化期间的房间。 </li><li>回到考场，打开活动室。即使数据不被收集在这个时候，这将进一步模拟测试环境。 </li><li>轻轻地从他们的家笼取出每只小鼠并立即将它们放置在试验室。如果活动室包含中央隔离带，分为 ​​室内到象限（ 图1），将一个鼠标到每个空象限。 </li><li>一旦所有的动物被装载到测试室中，将盖子上的每个测试室的顶部。留在该驯化时间的房间。 </li><li>在60分钟，回到房间。从各试验室中取出盖，轻轻将每只小鼠返回到他或她的各自家笼。 </li><li>清理各室用消毒液和纸巾。确保无灰尘颗粒被留在室内。 注意：如果多个会话正在运行的每一天，彻底清洁每个试验室中的每个会话之间。 </li><li>重复步骤1-6，连续4天。 注：执行驯化前一周初始数据收集。如果动物在整个研究中测试多次，仅先于第一轮测试，以避免习惯化进行驯化。此外，随机分配的动物到一个新的中的每个会话。跟踪框分配在整个研究的持续时间。 </li></ol> 2，数据收集<ol><li>将小鼠在试验室在他们的家笼10-30分钟，以适应新环境。留在此期间，该室。 </li><li>继10-30分钟，回到考场。打开活动室和连接到所述腔室中的计算机上打开伴随计算机软件。如果该腔室包含一个象限除法器，插入，此时的分区。 注意：如果该测试腔包含象限除法，两种动物可以在数据采集期间被放置在试验室中。一种动物可以被放置在由前左象限以及一个在背右象限（ 图1）。 注：数据收集过程中或在同一行或列中不要将动物在所有四个象限。将所述动物在这些取向将与红外光束的XY网格和动物运动干扰将精确地测量。 </li><li>配置计算机软件来进行prebeam检查。这种配置将允许一个运行的预束检查以下的实验装置和动物的插入之前，进入测试室（见下文）。 注意：当prebeam检查运行中，计算机软件评估了在xy红外线光束的功能。例如，它可确定是否光电发射体和受体的阻断而不能在腔室中适当地检测运动。 </li><li>设置在计算机软件中的原始数据收集参数，收集数据的6到10分钟块（ 即收集，共60分钟的数据），然后输入相应的日期，文件名 ​​，和鼠标的ID号。 </li><li>一旦所有的参数设置，运行prebeam检查。如果一个腔室没有通过prebeam检查它是最有​​可能是由于中央象限除法或试验室的对准不良。如果发生这种情况，重新调整中央象限迪威明镜和试验室，直到传感器不再受阻，该系统规定的试验室已准备就绪。如果这不能解决问题，参考仪器手册。 </li><li>当所有测试室都准备好了，轻轻地从他或她的家笼移开鼠标，立即把他或她进入测试室。注意鼠标的ID，并确保它输入到计算机中的相匹配。 </li><li>一旦所有的动物被适当地加载到测试室中，将盖子上的每个腔室的顶部。然后，在电脑软件中选择相应的命令来启动数据收集。此时，分析仪和计算机软件将根据数据收集参数开始记录的活动水平。 </li><li>离开试验室的测试期间的剩余部分。 </li><li>在测试期间（ 即 60分钟后）后，立即返回考场。保存该数据，然后将每个动物返回其相对于香港专业教育学院笼子。 </li><li>清理各单位用消毒纸巾。 注意：如果多个会话正在运行的每一天，彻底清洁每个试验室中的每个会话之间。 </li><li>将数据导出到电子表格，然后退出软件程序。 </li><li>检查数据，以确保他们的记录。如果没有记录数据，或将动物睡在整个数据采集的时间段的全部，执行数据收集的额外的一天。 注意：所述的动物被认为是“睡眠”，如果它不在整个60分钟的测试时间移动。 </li><li>重复步骤2.1-2.12连续4天。 注：如果动物在整个研究期间，在多个时间点进行测试，不进行公开活动领域的测量超过每月一次，以避免习惯。此外，随机每个会话分配的动物到一个新的对话框。跟踪框分配在整个研究的持续时间。 </li></ol> 3，数据分析<ol><li>计算的平均水平的活性（单位），垂直活动（单位），总行驶距离（厘米），运动时间（秒），和每小鼠和组休息时间（秒）。计算机软件计算并通过对所述数据收集期间（ 即 60分钟）的报告的总水平的活性（单位），垂直活动（单位），总行驶距离（厘米），动作时间（秒），和其余的时间（秒）每只小鼠。计算平均值的每一个，在4天的数据集合的上述参数。 </li><li>之前执行任何统计分析，评估使用夏皮罗 - 威尔克测试数据的正常性，并检查使用格拉布的测试离群值。删除任何显著离群值（P &lt;0.05）。 </li><li>对于正态分布的数据，比较使用两种独立样本t检验或单因素ANOVA和事后检验与调整多重比较的p值取决于基团之间装置决于治疗组的总数量。 </li><li>对于非正态分布的数据，无论是采用Wilcoxon秩和检验或Kruskal-Wallis检验和秩和检验用得到的调整取决于治疗组的总人数比较多的p值组间比较中值。 </li></ol>

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作者什么都没有透露。

旷场活动测定是一种体内测定法，可以是有益的，以评估在神经肌肉病6,7,11-14的动物模型中的疾病进展和药效。如示于图2中 ，它提供了活性水平的普遍反映机车功能的评估。这是一个不同的措施，不是肌肉的力量，使之成为一个理想的起次要或者辅助观察指标的临床前药物研究执行。此外，它是一种临床上相关的15，非侵入性的量度，其可以在整个研究期间，可以进行多次。然而，行为和机车活性也受其它因素，以及（ 即实验者处理，环境条件，和认知）产生变异在旷场活动数据。本文的目标是提供一个良好的测试和标准协议，减少变异，并允许结果为compared在多个实验室，在我们的领域内提高翻译的希望。 这一措施的主要缺点是，它是高度可变的，由众多的外部因素的影响。但是，我们开发协议时，考虑到了这一点。我们评估了各种测试方案包括持续时间1-5天收集数据的。最后，我们确定之前收集数据进行文书驯化熟悉的动物用试验室环境和执行4天的数据收集的变化量显著减少的结果数据5。该协议的最初目的是评估在mdx小鼠模型的行为和机车活动水平;然而，该协议是最近确认在Dy2J动物模型，以及6。有人建议，该协议在您的实验室之前，在临床前试验用它每个动物模型中的标准化。 开放领域的活动由遗传背景17，性别18-20，18岁，和昼夜节律变化21。这需要相同的年龄，性别，和遗传背景的动物在同一时间进行评估。在规划阶段，心思缜密，应投入决定在什么年龄或年龄层开阔的活动水平进行评估。每只动物模型有其独特的疾病进展和机车和行为表型，其中变化的严重程度和由6,15年龄（ 图2和图3）。因此，为了确定临床和病理相关的时间点，以评估旷场活动的措施是重要的。需要每个治疗组中，以检测统计学差异显著动物的总数由动物模型，年龄和性别而变化，以及。因此，有关样本量的计算也应该在规划阶段进行，以determ的INE需要每个治疗组中的动物的总数来检测统计上显著差异。这些计算还应该考虑到，在研究中使用的考虑额外的措施的结果（ 例如 ，握力测量或组织学分析）。基于我们的功率计算，我们通常使用每个治疗组10-12动物。此外，特别要注意在何种对照菌株被用于本研究。有不正当的对照菌株在临床前研究中使用的倾向。例如，BL6小鼠经常被用来作为对照菌株对mdx小鼠;然而，mdx小鼠是在BL10背景。正如在图3中，BL10小鼠比BL6小鼠，这使得不可能进行比较MDX和BL6数据更加活跃。当进行与mdx小鼠的临床前研究，BL10小鼠应作为对照菌株。此外，如果研究正在与Dy2J小鼠中进行，BL6小鼠应该被用作对照OL压力。 小环境的变化也可以显著影响活动水平。这些包括照明，温度，湿度，气味，噪声和人类活动4,15。因此，它是非常重要的，测试中在温度和湿度受控的房间与非直接照明在每天5相同的时间执行。试验室应均匀分布到整个房间，并在直接照明或阴影或黑暗的角落5不放置。动物应该随机分配到他们的测试室的每一天，以减少整个房间变化的环境条件的影响，并且应使其适应试验室10-30分钟之前，收集数据。确保跟踪各动物的方块分配在整个研究期间，以确保箱/环境的任何影响被均等地分布在不同的治疗组中。个人装阿尼玛LS进入测试腔室和处理在整个研究期间，将动物应当不清楚治疗组及动物应变时可能的。在许多情况下，受影响的基因型是由相关的控制显着不同和致盲是不可能的。不过，个人应始终处理和未处理组之间的蒙蔽。此外，所有个人都应该离开房间数据收集过程中，以减少房间内的噪声和干扰，所有室应收集数据的每次会议后进行彻底清洗。这些动作将显著减少的变化的数据。要注意的是，动物是也非常容易适应15是很重要的。因此，可以认为，动物从测试室直接以下每天在60分钟的数据收集和旷场活动水平被除去来评价不超过每月一次以上。 移动的总距离和总动换货时间的测量往往是最敏感的开放领域活动的测量5。在Dy2J模型中，垂直活动测定往往是最敏感的旷场活动度量（ 图3）;然而，它可以是很难捕捉的更小的动物精确垂直活动的测量。例如，它可能是一个较小的动物会显示出饲养行为和传感器不会捕获它由于垂直传感器的高度。因此，我们建议测试动物不早于5周龄。它也可能是一种动物会睡在整个数据采集时段的持续时间。如果是这种情况，它是适当补充数据收集的额外的一天。最后，内盒的象限除法器或传感器的堵塞对准不良可能导致不准确的数据也是如此。因此，这是非常重要结束后进行传感器预检查，在测试前，并检查所有数据每个数据采集时段。 分析开放领域的活动数据的注意事项也应考虑。开放领域的活动数据有一种倾向，是不正常分布，并有异常值4。在此之前执行任何统计分析，我们的生物统计学家强烈建议检查数据的正常和异常值。如果数据不服从正态分布，应该考虑比较装置时，利用非参数检验。另外，所有数据应该由不知道什么治疗组是一个个体进行分析。 总体而言，开放领域活动的措施具有重要的优点：a）它既是运动和行为的活动，这是强烈的，但并不总是具有相关功能的机车进行全面评估; B）这是一个简单的措施来执行;三）需要在测试过程中没有动物处理; D）它是一个可以在整个STU的时间来进行一次以上非侵入性的措施DY; E）没有特殊的训练是需要进行测试; F）多动物可以在同一时间进行测试;和g），它是一种临床相关的测量结果5,16。然而，检测治疗时，请记住，其他的因素会影响动物的行为，并依次开放领域活动的测量。药物可有中枢神经系统和等宽体效应和行为也可以通过紧张的环境影响。其结果是，它可能难以区分是否改变机车或行为活动水平改变有关的肌肉功能，肌肉力量，或者是从药物副作用所致。因此，额外的功能，组织学和或分子检测应进行为好。这个标准化的协议也被成功地用于其它肌肉疾病4,17;然而，如在图3中看到的，试验性研究应进行初步评估在动物中测量的灵敏度模型。

动物模型已经学习关于疾病的机制是有用的，但他们在临床试验中预测疗效效用经常被1-3挑战。每年有无数的“有为”的临床前研究发表;然而，很少有建议的干预显示出积极的结果时，移动到临床试验。这些差异往往归因于出版偏见，过于乐观的结论，并设计和执行，导致不能再现的结果1-3临床前研究甚少。 在药物发展为神经肌肉障碍的当前进展，对于设计良好的临床前试验的需求日益增加。特别是，有必要进行严格的方法，可以进行规范化，双盲的方式，与验证，重现性好，可翻译结果的措施。由于先天性肌肉中的一员病协会，与要进行更严格的临床前研究的愿望，我们在这里分享我们的标准作业程序（SOP）的开放式现场活动。这之前的SOP进行了验证4，出版作为TREAT-NMD的标准作业程序为假肥大型肌营养不良症（DMD）的动物模型5的一部分。我们已经使用了这种方法，以表型和测试的许多药物的治疗效果在各种神经肌肉疾病的动物模型，包括LAMA2 DY-2J / J（Dy2J）小鼠动物模型中为先天性肌营养不良（CMD）6,7 。反过来，这篇文章改编自我们之前发表的TREAT-NMD的SOP 5。 旷场活动监视系统全面评估运动和小鼠的行为学活动的水平，这可与机车的功能相关联。该试验也被广泛用于评估焦虑等与探索行为8-10。具体而言，开放领域是屁股一个有用的工具essing在神经肌肉病11,12的动物模型机车减值和治疗药物可以改善运动和/或运动功能-6,7,13,14的功效。开放领域的活动的评估提供了不同的措施，不是肌肉的力量，这就是通常与握力测量，它显示了如何药物会影响身体其他系统（如中枢神经系统），以及5。此外，开放领域活动的措施，总行驶距离，反映了6分钟步行试验，临床试验结果的措施，其重点是亚极量运动性能和寿命15,16质量。总的来说，这使得开放式现场活动测试在临床前试验中使用有益起次要或者辅助的测量结果。然而，在旷场活动的监控系统还具有与其相关联显著挑战。该测试是行为和因为它是由众多的外部影响可能相当可变因素。举例来说，这种行为可以通过探索驱动器（ 即认知），焦虑，疾病，生理节律，环境因素，遗传背景的影响，除了电机输出10。因此，它必须与一个受控的环境中进行标准化的方式这项措施。这里介绍的协议，详细描述了我们开放的领域活动的SOP。它提供了一步一步的过程和重要的考虑因素的进一步讨论，以控制环境条件，并有助于减少更详细的变化，典型的结果，数据分析和评估的优势和劣势。

<table border="0" cellpadding="0" cellspacing="0" width="722">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:184px;">Company</td>
			<td style="width:155px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Equipment</td>
			<td style="width:184px;"></td>
			<td style="width:155px;"></td>
			<td></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">VersaMax Open Field Activity Monitoring system with acrylic test chambers, and X, Y, Z axis sensors</td>
			<td style="width:184px;">AccuScan Instruments, Inc. Columbus Ohio, USA</td>
			<td style="width:155px;"></td>
			<td style="width:167px;">Retired</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Fusion Open Field Activity Monitoring system with acrylic test chambers, and X, Y, Z axis sensors</td>
			<td style="width:184px;">Omnitech Electronics, Inc. Columbus Ohio, USA</td>
			<td style="width:155px;"></td>
			<td style="width:167px;">Suggested system currently on the market</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Computer</td>
			<td style="width:184px;">Dell, Inc.&#160;</td>
			<td style="width:155px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Materials</td>
			<td style="width:184px;"></td>
			<td style="width:155px;"></td>
			<td></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Virkon-S Broad spectrum disinfectant (Potassium Peroxymonosulfate/ Sodium Chloride)</td>
			<td style="width:184px;">Pharmacal Research Laboratories, Inc.</td>
			<td style="width:155px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Mice</td>
			<td style="width:184px;"></td>
			<td style="width:155px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">B6.WK-Lama2dy-2J/J (Dy2J)</td>
			<td style="width:184px;">Jackson Lab</td>
			<td style="width:155px;">000524</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">C57BL/6J (BL6)</td>
			<td style="width:184px;">Jackson Lab</td>
			<td style="width:155px;">000664</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">SJL/J (SJL)</td>
			<td style="width:184px;">Jackson Lab</td>
			<td style="width:155px;">000686</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">C57BL/10ScSn-Dmdmdx/J (mdx)</td>
			<td style="width:184px;">Jackson Lab</td>
			<td style="width:155px;">001801</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">C57BL/10ScSnJ (BL10)</td>
			<td style="width:184px;">Jackson Lab</td>
			<td style="width:155px;">000476</td>
			<td style="width:167px;"></td>
		</tr>
	</tbody>
</table>

behavioral and locomotor measurements using an open field activity monitoring system for skeletal muscle diseases

露地活动监控系统全面评估运动和小鼠行为活动水平。它是评估神经肌肉疾病和治疗药物可以改善运动和/或肌肉功能疗效的动物模型机车减值一个有用的工具。露地酶活性的测定提供了不同的措施，不是肌肉的力量，这通常是由握力测量评估。它也可以显示如何药物可能会影响身体其他系统，以及额外的观察指标使用时。此外，如行驶总距离的措施反映了6分钟步行试验，临床试验的测量结果。然而，开放场活动监控也与显著挑战相关联的：打开字段活性测定根据动物的应变，年龄，性别，和昼夜节律变化。此外，房间温度，湿度，照明，噪音，甚至气味会影响评估的结果。总体来说，这款米anuscript提供了一个完善的测试和标准化的开放领域活动的SOP在神经肌肉疾病的动物模型的临床前试验。我们提供了重要的考虑因素，典型的结果，数据分析和详细的优势和开放现场测试的弱点进行了讨论。此外，我们在临床前试验用开放的领域活动时，为最佳的研究设计提供建议。

在分析开放领域的活动数据，我们专注于一些特定的测量提供活动水平的普遍反映机车功能的评估。这些参数包括：水平活动，垂直活动，运动时间，休息时间和行驶总距离。在一般情况下，动物具有降低的肌肉功能将不太活跃并具有较低的活动能力。这通常伴有下降水平活动，垂直活动，总行驶距离和运动时间，并增加休息时间5,6,12,17。相反，动物与未损伤的肌肉功能或那些具有降低恶化的肌肉病理的进展疗法治疗更有可能显示较高的活性水平6,7,14,17。 要显示的典型结果使用该协议在神经肌肉疾病的动物模型中获得的一个例子，我们从纵向研究瓦特提供的数据Ë以前在Dy2J先天性肌营养不良（CMD）的动物模型6进行。总之，Dy2J模型包含LAMA2基因导致后肢麻痹，脱髓鞘和营养不良骨骼肌变化的截短形式。这种肌肉病理上的活动水平的影响在这些小鼠中观察到。例如，Dy2J小鼠在研究中往往表现出较低水平的活动水平，以及更少的行驶距离，相比于他们的年龄和性别匹配的BL / 6野生型对照组在整个研究期间（ 图2）;然而，这些差异并非总是显著。意义的缺乏很可能是由于样本量小，并在BL6数据高集团内的变化。变化是典型的开放式现场活动数据;然而，特别是缺乏足够的电力这些数据，以确定是否这些基团是彼此显着差异。一个典型的n = 10-12，应使用以检测štatistically显著差异5,17。例如，当一个更大的样本大小的情况下，作为被在SJL研究完成（见第二BL6棒在图3B，3D和3E）组之间显著差异可以被观察到。该Dy2J动物也表现出完全丧失垂直活动水平，这反映了他们的后肢麻痹，并反过来，无能后方（ 图2B）6。最后，要注意的活性水平的性别差异是很重要的。例如，女性往往比男性更活跃，显示更高水平的横向活动，垂直活动和总行驶距离（ 图2）;然而，这些差异无统计学显著。 我们还从在神经肌肉疾病的其他动物模型中进行了多次先前的研究提供的数据，以及要强调的是影响行为几个其他因素ivity水平（ 图3）。例如，活动水平各不相同遗传背景的10。 BL10野生型小鼠表现出更大的横向活动，垂直活动和旅行相比年龄和性别匹配的BL6野生型小鼠（ 图3）的总距离。这是需要注意的，作为利用不正确的对照菌株的研究，可以使数据不可用的重要观察。第二，活动水平变化由疾病模型和表型（ 图3）。例如，在SJL小鼠的动物模型的肢带型肌营养不良-2B（LGMD-2B），示出了水平的活性和总行驶距离的最低水平，其次是Dy2J鼠标和mdx小鼠的动物模型为杜兴肌营养不良症（ 图3A，图3C）。然而，Dy2J小鼠，因为它们的后肢麻痹，显示垂直活性（ 图3B）的最低水平。同样重要的是要注意，在较高的水平在MDX表型活性最可能归因于BL10背景应变的增加的活性水平。最后，这一数字凸显在测试时动物的年龄/疾病病理的重要性。例如，在30周龄时，没有差异，可以在mdx小鼠和它们的年龄和性别匹配的BL10野生型对照（ 图3）之间的活动水平进行检测。然而，在6周龄时，围绕在mdx小鼠模型中的峰坏死的相位，mdx小鼠显示显著减少垂直活动和水平活动。的降低，也观察到在行进的总距离，但这种差异并不显著（ 图3）4。 <img alt="图1" fo:content-width="5in" src="/files/ftp_upload/51785/51785fig1highres.jpg" width="500" /> 图1：打开现场活动的设备。两个领域的开放装置与中心QUADRANT分频器，和上衣。如果可移动的中央分隔象限存在，动物应该只（4 2，）每箱在测试过程中获得有效的读数放在左前方（1,3）和右后室。 <img alt="图2" fo:content-width="5in" src="/files/ftp_upload/51785/51785fig2highres.jpg" width="500" /> 图2：打开字段中的活动数据。为Dy2J典型的开放式现场活动数据（灰线，N = 3），年龄和性别匹配的BL6对照菌株（黑线，N = 3）的小鼠在14，19，23，25，和年龄30周（A - F）A）水平活动（任意单位）的男性，B）水平活动（任意单位）女，C）的垂直活动（任意单位）的男性，D）垂直活动（任意单位）的女性。 E）的总距离特拉沃LED（厘米）的男性，E）的总行驶距离（厘米）女性。收集在连续4天的数据，每个鼠标和组平均值。相同的小鼠在各时间点进行测试。数据表示平均值±标准差。数据此前公布的6。 Dy2J和BL6小鼠用独立样本t检验在各时间点进行比较。 p值&lt;0.05为差异有显著。 * P &lt;0.05，** P &lt;0.01，*** P &lt;0.001。 <img alt="图3" fo:content-width="5in" src="/files/ftp_upload/51785/51785fig3highres.jpg" width="500" /> 图3：打开现场活动数据多株 BL10男性的典型行为活动数据（6周龄，每组8只，25〜30周龄时，N = 10），MDX（6周龄，N = 9; 25-30周龄，每组15只），BL6（对照组为Dy2J小鼠，n = 3时，对照组为SJL小鼠中，n = 13），Dy2J组（n = 3），和SJL组（n = 13）小鼠在VArying年龄A）水平活动（任意单位）由BL10和mdx小鼠的数据，在图6和25-30周龄，B）的水平活动（任意单位）从BL6，Dy2J和SJL小鼠数据在25-30周的年龄，C），垂直活动（任意单位）由BL10和mdx小鼠的数据，在图6和25-30周龄，D）垂直活动（任意单位）从BL6，Dy2J和SJL小鼠数据在25-30周龄时，E）总行驶距离（厘米）从BL10和mdx小鼠在6和25〜30周龄，和F）的总的行驶距离数据（厘米）从BL6，Dy2J和SJL小鼠数据在25-30周龄时。 Dy2J，对于CMD动物模型上的BL6背景层粘连蛋白α2基因突变; SJL，动物模型肢带型肌营养不良-2B（LGMD-2B）; MDX，对于DMD动物模型上的BL10背景。没有dysferlin-足以SJL控制。数据是平均值±SEM。 B，C和D包含数据预先PUBLI棚6,17。数据不是正态分布;因此，数据是用Wilcoxon秩款项检验。 P &lt;0.05的p值被认为显著。下面的比较为每个参数被制成：1）BL10和mdx小鼠在6周龄时，P &lt;0.05;二）BL10和mdx小鼠在25-30周龄时，不显著;三）Dy2J和BL6在25-30周龄时，P &lt;0.05相匹配的对照菌株的小鼠;四）SJL和BL6在25-30周龄时，P &lt;0.001相匹配的对照菌株;五）BL10小鼠在25-30周龄和BL6（对照组为Dy2J小鼠）的小鼠在25周龄时，P &lt;0.01;六）BL10在25-30周龄和BL6（对照组为SJL小鼠）在25-30周龄时，P &lt;0.001。

Watch this Scientific Journal Video about Behavioral and Locomotor Measurements Using an Open Field Activity Monitoring System for Skeletal Muscle Diseases at JoVE.com

Behavioral and Locomotor Measurements Using an Open Field Activity Monitoring System for Skeletal Muscle Diseases

开放领域的活动水平来评估机车和行为活动水平。该协议提供了一个精心设计的，标准化的协议为神经肌肉疾病的临床前试验中使用。

行为和运动测量使用一个开放的领域活动监控系统的骨骼肌疾病

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing ...

behavioral-locomotor-measurements-using-an-open-field-activity

Research

JoVE Journal

Behavior

53.3K 次观看.  Children's National Medical Center. 开放领域的活动水平来评估机车和行为活动水平。该协议提供了一个精心设计的，标准化的协议为神经肌肉疾病的临床前试验中使用。 

视频: Behavioral and Locomotor Measurements Using an Open Field Activity Monitoring System for Skeletal Muscle Diseases

Mouse Short- and Long-term Locomotor Activity Analyzed by Video Tracking Software

Locomotor activity (LMA) is a simple and easily performed measurement of behavior in mice and other rodents. Improvements in video tracking software (VTS) have allowed it to be coupled to LMA testing, dramatically improving specificity and sensitivity when compared to the line crossings method with manual scoring. In addition, VTS enables high-throughput experimentation. While similar to automated video tracking used for the open field test (OFT), LMA testing is unique in that it allows mice to remain in their home cage and does not utilize the anxiogenic stimulus of bright lighting during the active phase of the light-dark cycle. Traditionally, LMA has been used for short periods of time (mins), while longer movement studies (hrs-days) have often used implanted transmitters and biotelemetry. With the option of real-time tracking, long-, like short-term LMA testing, can now be conducted using videography. Long-term LMA testing requires a specialized, but easily constructed, cage so that food and water (which is usually positioned on the cage top) does not obstruct videography. Importantly, videography and VTS allows for the quantification of parameters, such as path of mouse movement, that are difficult or unfeasible to measure with line crossing and/or biotelemetry. In sum, LMA testing coupled to VTS affords a more complete description of mouse movement and the ability to examine locomotion over an extended period of time.

Locomotor activity (LMA) is a simple and easily performed measurement of behavior in mice. Coupling of video tracking software (VTS) and LMA allows for the improvement of specificity and sensitivity, especially when compared with the manual, line crossing method of LMA analysis. Additionally VTS allows long-term tracking of mouse LMA.

鼠标短期和长期活动性分析视频跟踪软件

Locomotor  activity (LMA) is a simple and easily performed measurement of behavior in mice  and other rodents. Improvements in video tracking software ...

科研

行为学

Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of the most commonly used platforms to measure behaviors in animal models. It is a fast and relatively easy test that provides a variety of behavioral information ranging from general ambulatory ability to data regarding the emotionality of the subject animal. As it relates to rodent models, the procedure allows the study of different strains of mice or rats both laboratory bred and wild-captured. The technique also readily lends itself to the investigation of different pharmacological compounds for anxiolytic or anxiogenic effects. Here, a protocol for use of the open field maze to describe mouse behaviors is detailed and a simple analysis of general locomotor ability and anxiety-related emotional behaviors between two strains of C57BL/6 mice is performed. Briefly, using the described protocol we show Wild Type mice exhibited significantly less anxiety related behaviors than did age-matched Knock Out mice while both strains exhibited similar ambulatory ability.

A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.

使用开放式现场迷宫来衡量小鼠的运动能力和焦虑样行为

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of ...

Place and Response Learning in the Open-field Tower Maze

This protocol describes how the Open-field Tower Maze (OFTM) paradigm is used to study spatial learning in rodents. This maze is especially useful for examining how rats learn to use a place- or response-learning to successfully navigate in an open-field arena. Additionally, this protocol describes how the OFTM differs from other behavioral maze paradigms that are commonly used to study spatial learning in rodents. The OFTM described in this article was adapted from the one previously described by Cole, Clipperton, and Walt (2007). Specifically, the OFTM was created to test spatial learning in rodents without the experimenter having to consider how &#8220;stress&#8221; might play a role as a confounding variable. Experiments have shown that stress-alone can significantly affect cognitive function1. The representative results section contains data from an experiment that used the OFTM to examine the effects of estradiol treatment on place- and response-learning in adult female Sprague Dawley rats2. Future studies will be designed to examine the role of the hippocampus and striatum in place- and response-learning in the OFTM.

The Open-field Tower Maze (OFTM) is a maze created to study the behavioral and neural mechanisms of spatial learning (e.g., place- or response-learning) in rats. This maze is especially useful for experimenters who want to use a non-stressful maze paradigm to investigate spatial learning in their research.

在打开场塔迷宫的地方，反应学习

This protocol describes how the Open-field Tower Maze (OFTM) paradigm is used to study spatial learning in rodents. This maze is especially useful for ...

Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria

Captive-rearing observations are valuable for revealing aspects of fish behavior and ecology when continuous field investigations are impossible. Here, a series of basic techniques are described to enable observations of the reproductive behavior of a wild-caught gobiid fish, as a model, kept in an aquarium. The method focuses on three steps: collection, transport, and observations of reproductive ecology of a substrate spawner. Essential aspects of live fish collection and transport are (1) preventing injury to the fish, and (2) careful acclimation to the aquarium. Preventing harm through injuries such as scratches or a sudden change of water pressure is imperative when collecting live fish, as any physical damage is likely to negatively affect the survival and later behavior of the fish. Careful acclimation to aquaria decreases the incidence death and mitigates the shock of transport. Observations during captive rearing include (1) the identification of individual fish and (2) monitoring spawned eggs without negative effects to the fish or eggs, thereby enabling detailed investigation of the study species' reproductive ecology. The subcutaneous injection of a visible implant elastomer (VIE) tag is a precise method for the subsequent identification of individual fish, and it can be used with a wide size range of fish, with minimal influence on their survival and behavior. If the study species is a substrate spawner that deposits adhesive eggs, an artificial nest site constructed from polyvinyl chloride (PVC) pipe with the addition of a removable waterproof sheet will facilitate counting and monitoring the eggs, lessening the investigator's influence on the nest-holding and egg-guarding behavior of the fish. Although this basic method entails techniques that are seldom mentioned in detail in research articles, they are fundamental for undertaking experiments that require the captive rearing of a wild fish.

A series of basic methods to enable the study of the reproductive ecology of fish kept in aquaria are described. These are useful protocols for collecting fish using SCUBA, transporting live fish, and observing the reproductive behavior of wild-caught fish kept in aquaria.

鱼类生殖生态学研究的基本方法

Captive-rearing observations are valuable for revealing aspects of fish behavior and ecology when continuous field investigations are impossible. Here, a ...

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 ...

Automated Measurements of Sleep and Locomotor Activity in Mexican Cavefish

Across phyla, sleep is characterized by highly conserved behavioral characteristics that include elevated arousal threshold, rebound following sleep deprivation, and consolidated periods of behavioral immobility. The Mexican cavefish, Astyanax mexicanus (A. mexicanus), is a model for studying trait evolution in response to environmental perturbation. A. mexicanus exist as in eyed surface-dwelling forms and multiple blind cave-dwelling populations that have robust morphological and behavioral differences. Sleep loss has occurred in multiple, independently-evolved cavefish populations. This protocol describes a methodology for quantifying sleep and locomotor activity in A. mexicanus cave and surface fish. A cost-effective video monitoring system allows for behavioral imaging of individually-housed larval or adult fish for periods of a week or longer. The system can be applied to fish aged 4 days post fertilization through adulthood. The approach can also be adapted for measuring the effects of social interactions on sleep by recording multiple fish in a single arena. Following behavioral recordings, data is analyzed using automated tracking software and sleep analysis is processed using customized scripts that quantify multiple sleep variables including duration, bout length, and bout number. This system can be applied to measure sleep, circadian behavior, and locomotor activity in almost any fish species including zebrafish and sticklebacks.

This protocol details the methodology for quantifying locomotor behavior and sleep in the Mexican cavefish. Previous analyses are extended to measure these behaviors in socially-housed fish. This system can be widely applied to study sleep and activity in other fish species.

墨西哥河鱼睡眠和睡眠活动的自动测量

Across phyla, sleep is characterized by highly conserved behavioral characteristics that include elevated arousal threshold, rebound following sleep ...

Using Facial Electromyography to Assess Facial Muscle Reactions to Experienced and Observed Affective Touch in Humans

"Affective" touch is believed to be processed in a manner distinct from discriminatory touch and to involve activation of C-tactile (CT) afferent fibers. Touch that optimally activates CT fibers is consistently rated as hedonically pleasant. Patient groups with impaired social-emotional functioning also show disordered affective touch ratings. However, relying on self-reported ratings of touch has many limitations, including recall bias and communication barriers. Here, we describe a methodological approach to study affective responses to touch via facial electromyography (EMG) that circumvents the reliance on self-report ratings. Facial EMG is an objective, quantitative, and non-invasive method to measure facial muscle activity indicative of affective responses. Responses can be assessed across healthy and patient populations without the need for verbal communication. Here, we provide two separate datasets demonstrating that CT-optimal and non-optimal touch elicit distinct facial muscle reactions. Moreover, facial EMG responses are consistent across stimulus modalities, e.g. tactile (experienced touch) and visual (observed touch). Finally, the temporal resolution of facial EMG can detect responses on timescales that supersede that of verbal reporting. Together, our data suggest that facial EMG is a suitable methodology for use in affective tactile research that can be used to supplement, or in some cases, supplant, existing measures.

We describe a protocol to assess facial muscle activity in response to experienced and observed tactile stimulation using facial electromyography.

面部肌电图对人体有经验和观察到的情感触觉的影响评估

"Affective" touch is believed to be processed in a manner distinct from discriminatory touch and to involve activation of C-tactile (CT) afferent fibers. ...

Flypub To Study Ethanol Induced Behavioral Disinhibition and Sensitization

Alcohol use disorder (AUD) remains a serious problem in our society. To develop effective interventions for addiction, it is important to understand the underlying neurobiological mechanisms, for which diverse experimental approaches and model systems are needed. The main ingredient of alcoholic beverages is ethanol, which causes adaptive changes in the central nervous system and behavior upon chronic intake. Behavioral sensitization (i.e., escalated responses) in particular represents a key adaptive change underlying addiction. Most ethanol-induced behavioral sensitization studies in animal models have been conducted on the locomotor activating effect of ethanol. A prominent effect of ethanol is behavioral disinhibition. Behavioral sensitization to the disinhibition effect of ethanol, however, is underrepresented. To address this issue, we developed the Flypub assay that allows measuring the escalated increase in disinhibited courtship activities upon recurring ethanol exposure in Drosophila melanogaster.&#160;Here, we report the step-by-step Flypub assay including assembly of ethanol exposure chambers, setup of the assay station, criteria for fly care and collection, ethanol delivery, quantification of disinhibited courtship activities, data processing and statistical analysis. Also provided are how to troubleshoot critical steps, overcome limitations and expand its utility to assess additional ethanol-induced behaviors. The Flypub assay in combination with powerful genetic tools in Drosophila melanogaster will facilitate the task of discovering the mechanism underlying ethanol-induced behavioral sensitization.

The Flypub assay measures the behaviors that the fruit fly Drosophila melanogaster displays under the influence of ethanol. The assay can be readily mastered by experimenters at all levels and applied to various vaporized stimuli, facilitating substance abuse and addiction studies.

飞行酒吧研究乙醇诱导的行为抑制和敏感性

Alcohol use disorder (AUD) remains a serious problem in our society. To develop effective interventions for addiction, it is important to understand the ...

An Adjustable High-Definition Imaging System for Behavioral Studies of Drosophila Adults

Drosophila melanogaster is a very powerful model in biological research, but a bad model for photography or videography. This paper describes a simple but effective method to observe and document the behavior or morphology of flies. Flies were placed in a translucent observation chamber c.a. &#216;15 x 5mm (no food inside) or &#216;15 x 12 mm (with an 8 mm-high piece of food inside). After covering with an ultraviolet (UV)/clear filter with high light transmittance, the chamber was placed under a 5-50x zoom stereo microscope, and mini light-emitting diode (LED) video lights were placed on both sides of the microscope to illuminate the chamber to obtain uniform, soft, bright, and nearly shadow-free light. Then, a compact digital camera with 3-5x optical zoom, which can record 1080 P high-definition or higher resolution video (at a frame rate of &#8805;30 fps), was connected to the eyepiece of microscope through a bracket, and photographs or videos were taken through the eyepiece. By adjusting the zoom knob of the zoom stereo microscope, it was very easy to track the flies and take panoramic or detailed close-up images as needed, while the camera recorded everything under the microscope. Because the flies can stay at any position in the chamber, they can be observed and recorded from all directions. The photographs or videos taken are of good image quality. This method can be used both for scientific research and teaching.

An Adjustable High-Definition Imaging System for Behavioral Studies of Drosophila Adults

This protocol describes how to make a simple adult Drosophila behavior observation chamber, and how to take high-definition photographs/videos of the morphology or behavior of different types of fruit flies in the observation chamber through relatively simple and affordable methods.

可调高清成像系统，用于 果蝇 成人的行为研究

Drosophila melanogaster is a very powerful model in biological research, but a bad model for photography or videography. This paper describes a simple but ...

An Open-Source, Fully Customizable 5-Choice Serial Reaction Time Task Toolbox for Automated Behavioral Training of Rodents

The 5-choice serial reaction time task (5-CSRTT) is a behavioral test often used to study visuospatial attention and impulsiveness in rodents. The task requires animals to allocate attention to a horizontal array of five small apertures equipped with light sources and, within a limited time window, nose-poke one illuminated target aperture to get a food reward at the food magazine located in the opposite wall of the chamber. The task considers behavioral control measures such as response accuracy and reaction times and allows to infer selective attention and impulsivity. Task difficulty can be controlled by modifying the stimulus duration and task design in general. Commercially available apparatus usually consists of an experimental chamber and particular software to specify task parameters, but due to fixed hard- and software, they pose many limitations on changes in the general experimental design and specific task requirements and the related data output. This article explains a fully customizable alternative based on an easy-to-use single-board microcontroller and standard electrotechnical components, an open-access Arduino script, and a Matlab-toolbox for hardware control and behavioral task specifications, respectively. The toolbox includes an optional staircase procedure, enabling automated behavioral training. The complete hardware setup, which can be installed in customized chambers, and the freely adaptable software encourage non-standardized task and chamber design. The design of the system and the open-source code for hardware control and experimental setup are described.

The present protocol describes the development of an open-source 5-choice serial reaction time task toolbox for rodent animal models, using Arduino and related hardware and a versatile Matlab toolbox, including an optional script for automated behavioral training. The scripts are customizable and facilitate the implementation of different trial-and-test designs.

一个开源的，完全可定制的5选择串行反应时间任务工具箱，用于啮齿动物的自动行为训练

The 5-choice serial reaction time task (5-CSRTT) is a behavioral test often used to study visuospatial attention and impulsiveness in rodents. The task ...