Name: a simple and low-cost assay for measuring ambulation in mouse models of muscular dystrophy
Uploaded: 2017-12-29T14:00:00
Description: Watch this Scientific Journal Video about A Simple and Low-cost Assay for Measuring Ambulation in Mouse Models of Muscular Dystrophy at JoVE.com

这项工作得到了国家卫生研究院的资助 [R01 AR048179 和 R01 HL126204 到钢筋混凝土 W, T32 AR059033 和 F32 AR069469to 福斯特 G] 和肌肉萎缩症协会美国 [274143 和416364至 R.C.W.]。

这里描述的方法得到了加州大学洛杉矶分校动物保育和使用委员会的批准。野生类型 (C57BL/6J) 和mdx小鼠 (C57Bl/10ScSn 背景) 是从商业来源购买的。mdx: utrophin-Tg 小鼠是来自 Ervasti 的礼物, 得到了凯戴维斯的许可, 并在mdx背景上进行了维护。6月龄雄性小鼠进行了步行试验。老鼠在 Terasaki 生命科学 Vivarium 被维护了由动物看护和用途委员会建立的指南在加利福尼亚大学洛杉矶 (批准 #2000...

<ol>
	<li>Nagaraju, K., Carlson, G., De Luca, 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+locomotor+measurements+using+open+field+animal+activity+monitoring+system.">Behavioral and locomotor measurements using open field animal activity monitoring system.</a> TREAT-NMD SOP Number M2.1.002. 2. 2, (2010).</li><li>Tatem, K. S., 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=Behavioral+and+locomotor+measurements+using+an+open+field+activity+monitoring+system+for+skeletal+muscle+diseases.">Behavioral and locomotor measurements using an open field activity monitoring system for skeletal muscle diseases.</a> J Vis Exp. (91), e51785 (2014).</li><li>Kobayashi, Y. M., 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=Sarcolemma-localized+nNOS+is+required+to+maintain+activity+after+mild+exercise.">Sarcolemma-localized nNOS is required to maintain activity after mild exercise.</a> Nature. 456 (7221), 511-515 (2008).</li><li>Belanto, J. 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=Microtubule+binding+distinguishes+dystrophin+from+utrophin.">Microtubule binding distinguishes dystrophin from utrophin.</a> Proc Natl Acad Sci U S A. 111 (15), 5723-5728 (2014).</li><li>McDonald, C. M., 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=The+6-minute+walk+test+in+Duchenne/Becker+muscular+dystrophy:+longitudinal+observations.">The 6-minute walk test in Duchenne/Becker muscular dystrophy: longitudinal observations.</a> Muscle Nerve. 42 (6), 966-974 (2010).</li><li>Mazzone, 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=Star+Ambulatory+Assessment,+6-minute+walk+test+and+timed+items+in+ambulant+boys+with+Duchenne+muscular+dystrophy.">Star Ambulatory Assessment, 6-minute walk test and timed items in ambulant boys with Duchenne muscular dystrophy.</a> Neuromuscul Disord. 20 (11), 712-716 (2010).</li><li>Kobayashi, Y. M., Rader, E. P., Crawford, R. W., Campbell, K. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endpoint+measures+in+the+mdx+mouse+relevant+for+muscular+dystrophy+pre-clinical+studies.">Endpoint measures in the mdx mouse relevant for muscular dystrophy pre-clinical studies.</a> Neuromuscul Disord. 22 (1), 34-42 (2012).</li><li>Acosta, A. R., 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=Use+of+the+six-minute+walk+test+to+characterize+golden+retriever+muscular+dystrophy.">Use of the six-minute walk test to characterize golden retriever muscular dystrophy.</a> Neuromuscul Disord. 26 (12), 865-872 (2016).</li><li>Sousa, N., Almeida, O. F., Wotjak, C. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+hitchhiker's+guide+to+behavioral+analysis+in+laboratory+rodents.">A hitchhiker's guide to behavioral analysis in laboratory rodents.</a> Genes Brain Behav. 5, 5-24 (2006).</li><li>Gibbs, E. M., 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=High+levels+of+sarcospan+are+well+tolerated+and+act+as+a+sarcolemmal+stabilizer+to+address+skeletal+muscle+and+pulmonary+dysfunction+in+DMD.">High levels of sarcospan are well tolerated and act as a sarcolemmal stabilizer to address skeletal muscle and pulmonary dysfunction in DMD.</a> Hum Mol Genet. 25 (24), 5395-5406 (2016).</li><li>Gillis, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multivariate+evaluation+of+the+functional+recovery+obtained+by+the+overexpression+of+utrophin+in+skeletal+muscles+of+the+mdx+mouse.">Multivariate evaluation of the functional recovery obtained by the overexpression of utrophin in skeletal muscles of the mdx mouse.</a> Neuromuscul Disord. 12, S90-S94 (2002).</li><li>Tinsley, 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=Expression+of+full-length+utrophin+prevents+muscular+dystrophy+in+mdx+mice.">Expression of full-length utrophin prevents muscular dystrophy in mdx mice.</a> Nat Med. 4 (12), 1441-1444 (1998).</li><li>Song, 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=Suite+of+clinically+relevant+functional+assays+to+address+therapeutic+efficacy+and+disease+mechanism+in+the+dystrophic+mdx+mouse.">Suite of clinically relevant functional assays to address therapeutic efficacy and disease mechanism in the dystrophic mdx mouse.</a> J Appl Physiol. 122 (3), 593-602 (2017).</li><li>Bolivar, V. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrasession+and+intersession+habituation+in+mice:+from+inbred+strain+variability+to+linkage+analysis.">Intrasession and intersession habituation in mice: from inbred strain variability to linkage analysis.</a> Neurobiol Learn Mem. 92 (2), 206-214 (2009).</li></ol>

野外试验提供了一种有效而非侵入性的方法来检测肌肉疾病的小鼠模型中的运动功能, 而这种检测的变异可以作为临床前研究的一项重要的终点措施。这类化验的一个主要限制是活动监测系统的高成本和有限的可及性。在这项研究中, 我们展示了一个低成本的开放现场活动分析系统, 可以产生类似的结果, 现有的商业软件和设备。该系统可以在 vivaria 的地方使用, 从而避免了将动物转移到核心设施的...

可靠和可重复的测量肌肉功能是关键的评估潜在的治疗 DMD 的疗效。DMD 是由肌营养不良蛋白基因突变引起的遗传性疾病, 导致渐进性肌肉无力, 移动丧失, 最终心肺衰竭。最广泛使用的 DMD 的动物模型是肌营养不良蛋白-空mdx鼠标。在mdx鼠标以及其他营养和病的类似动物模型中, 一组功能测试已成为评估疾病进展的常规方法。通常使用的在体内测试包括测量前肢握力, 导线悬挂时间, rotarod 最大, 在跑步机跑步的时间和运动活动跟踪的疲劳。在这一领域, 为了使这些测试标准化, 有了很大的努力, 目的是减少前临床研究的变异性, 提高在小鼠体内测试的治疗的转化潜能1,2。
临床前测试的一个重要类别是测量自愿运动, 这一参数在小鼠肌肉萎缩症模型中经常被改变。这通常是通过基于野外活动监测的分析来测试的, 并且可以在几分钟或数小时的过程中评估水平 (行走) 或垂直 (饲养) 运动,2,3,4。一些研究表明, 在mdx小鼠中, 特别是在运动之后, 自愿改变的运动, 这些测量结果对药物治疗和疾病进展很敏感。执行这些化验的一个主要限制是需要专门的、高成本的设备。在这里, 一个低成本的方法, 跟踪鼠标移动使用现成的资源。
6分钟步行距离是一种常用的标准作为临床评估工具的个人杜氏肌营养不良症5,6。这项措施的修改已被用来评估杜氏动物模型的结果, 包括mdx小鼠7和金毛猎犬肌肉萎缩症 (GRMD) 狗8。在这项研究中, 我们记录自愿开放野外运动在6分钟后, 紧接着一个温和的运动挑战。然后使用自由开源软件计算移动距离, 以测量随时间推移的水平移动。
这种方法的主要优点是可以在不同的环境中测试动物, 而无需专门的设备或昂贵的商业软件进行分析。这一分析的一个重要方面是, 它可以在一个基本的实验室设置, 而无需移动或转移到一个专门的核心设施 vivarium 小鼠。这里描述的视频跟踪协议非常适合于对相对较短的青年进行移动评估, 并且能够检测出野生类型和mdx小鼠之间的活动差异, 并揭示了救援中的功能改进模型的 DMD。

<table>
	<tbody>
		<tr>
			<td>Video camera</td>
			<td>Apple Inc.</td>
			<td>ME553LL/A</td>
			<td>For recording ambulation video, iPhone 5S Plus (or equivalent)</td>
		</tr>
		<tr>
			<td>Kinovea software (version 0.8.15)</td>
			<td>Kinovea Association</td>
			<td></td>
			<td>Open source video analysis software. Free download, PC compatible (Version 0.8.15, www.kinovea.org)</td>
		</tr>
		<tr>
			<td>iMovie (version 10.0.6)</td>
			<td>Apple Inc.</td>
			<td></td>
			<td>Any similar software can be used to reduce video frame rate (optional)</td>
		</tr>
		<tr>
			<td>Roughneck 32 Gallon Black Round Trash Bin (Open field chamber)</td>
			<td>Rubbermaid</td>
			<td># 1778013</td>
			<td>Any open field chamber system can be adapted for recording. This study uses a recording chamber constructed out of a tray on a platform, at the bottom of a large trash bin.</td>
		</tr>
		<tr>
			<td>Avant White Plastic Tray 15&#34;W x 10&#34;D x 1.45&#34;H (Open field chamber)</td>
			<td>US Acrylic, LLC</td>
			<td></td>
			<td>Any open field chamber system can be adapted for recording. This study uses a recording chamber constructed out of a tray on a platform, at the bottom of a large trash bin.</td>
		</tr>
		<tr>
			<td>C57BL/6J</td>
			<td>Jackson Laboratory</td>
			<td>#000664</td>
			<td>Male 6 month mice</td>
		</tr>
		<tr>
			<td>C57BL/10ScSn-Dmd/J (mdx)</td>
			<td>Jackson Laboratory</td>
			<td>#001801</td>
			<td>Male 6 month mice</td>
		</tr>
		<tr>
			<td>mdx: utrophin-Tg (fiona)</td>
			<td>Gift from from James Ervasti, with permission from Kay Davies</td>
			<td></td>
			<td>Male 6 month mice</td>
		</tr>
	</tbody>
</table>

a simple and low-cost assay for measuring ambulation in mouse models of muscular dystrophy

在肌营养不良症的治疗中, 测量功能性结局是临床前测试的一个重要方面。在小鼠模型中自愿移动的评估是一种非侵入性的和可重复的活性测定方法, 与患者步行6分钟的测量和相关的移动评分直接类似。许多测试鼠标移动速度和距离的常用方法都是基于野外试验, 在这个测试中, 动物在竞技场中的自由运动是随着时间的推移而测量的。这种方法的一个主要缺点是, 用于高分辨率运动跟踪的商用软件和设备成本高昂, 可能需要将鼠标转移到专门的设备进行测试。在这里, 我们描述了一个低成本, 视频的测量鼠标移动的系统, 利用自由和开源软件。使用这个协议, 我们证明, 自愿移动在肌营养不良蛋白-无效的假mdx小鼠模型的杜氏肌肉萎缩症 (DMD) 是减少相对于野生类型的鼠标活动。在表达 utrophin 转基因的mdx小鼠中, 没有观察到这些活动的缺陷, 并且总的移动距离与野生型小鼠是无法区分的。这种方法是有效的测量变化的自愿步行与不良病理学, 并提供了一个多才多艺的平台, 可以很容易地适应不同的研究设置。

开放野外试验的变化已被证明是一种有效的方法来测试疾病进展的模型肌肉萎缩症。在这里, 我们演示了一种通用的视频跟踪方法, 用于分析 two-dimensional 鼠标移动使用现成的记录设备和开源软件 (图 1A-B)。与类似分析报告的结果一致, 我们显示mdx小鼠在测试之前, 在一个6分钟的野外任务中, 比野生型小鼠更少的自愿运动...

Watch this Scientific Journal Video about A Simple and Low-cost Assay for Measuring Ambulation in Mouse Models of Muscular Dystrophy at JoVE.com

A Simple and Low-cost Assay for Measuring Ambulation in Mouse Models of Muscular Dystrophy

一种简便、低成本的肌营养不良小鼠步行模型的测定方法

该协议描述了一个灵活的, 低成本的测量鼠标移动在野外活动的测试系统。我们显示, 基于这个系统的6分钟的步行试验检测到在mdx小鼠中自愿运动的减少, 并准确地区分了这些动物的肌肉特异性抢救的改进。

Measuring functional outcomes in the treatment of muscular dystrophy is an essential aspect of preclinical testing. The assessment of voluntary ambulation ...

Research

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巴恩斯迷宫测试策略与小型和大型啮齿类动物模型

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