我们要感谢玛格丽特Hulsker为她照相援助和获得小鼠的图像和审稿人非常有建设性的意见帮助。这项工作是由ZonMw，TREAT-NMD（合同号利星行-CT-2006-036825）和杜氏父项目的支持。

这里所描述的实验是批准莱顿大学医学中心（LUMC）的动物伦理委员会（DEC）。小鼠经LUMC的动物饲养设施，并保持在单独通风笼12小时光线暗周期。他们有自由采食饮水和标准饲料。 当执行任何下文所述的功能测试，实验条件必须严格控制，以减少偏差。最好是，年龄和性别匹配的老鼠应该使用，年龄和性别之间的性能差异。属于同窝小鼠应该随机对实验组。动物应该由同一运营商，谁是失明的实验组进行测试。测试应在白天和平日，同室的同时进行均衡的气味，噪音等个别小鼠和时间点之间14大变化可以用于所有被观察功能测试，因此6-8只/实验组应该被使用。功能测试性能也可以在很大程度上不同的近交系野生型菌株之间的差异。因此，实验组和对照野生型小鼠一定要有相应的背景（在mdx小鼠的使用C57BL/10ScSnJ野生型菌株）。这里描述的所有数据已取得与C57BL/10ScSnJ野生型菌株，其我们称之为野生型从这里开始。这里所描述的测试可以使用纵向从MDX和野生型小鼠至少1-19个月的年龄。测试不应该被重复超过每周一次，以防止老鼠失去兴趣和意愿来执行任务。 1。前肢握力测试使用前肢握力测试来测量前肢的强度。该测试是基于鼠标的倾向，本能地抓住一个网格时的尾巴15暂停，ADAPTEd从<a href="http://www.treat-nmd.eu/downloads/file/sops/dmd/MDX/dmd_m.2.2_001.pdf" target="_blank">DMD_M2.2.001.pdf</a> 。 <ol><li>设备设置:将一个网格，力传感器，测量拉过程中应用通过鼠标对电网的最大力量。确保设置是山顶上的张力模式（T-PK）的拉动。力的单位可以在任一盎司 - 力的，克 - 力的，磅力的，公斤 - 力的，或牛顿进行调整。 注意:我们喜欢与克为单位的值来工作。多种米市售，但只有轴向传感器给出可靠的结果为杠杆式力传感器是通过杠杆效应的物理规律负面影响。无论是nonflexible网格或三角形可用于棒是1-2毫米的直径。 </li><li>在测试之前，评估体重的鼠标，从而使归一化对于体重。 </li><li>用克为单位的值。重置仪表在每个RECO开始录制。 </li><li>通过抓住尾巴和水平移动对电网从笼中取出鼠标。 </li><li>检查鼠标掌握电网紧密地与两个前爪。 </li><li>拖动鼠标远离电网，以便其掌握被打破，适用于电网的最高力量将换能器的显示，它可以是手动或自动记录上显示。 </li><li>只需要拉考虑在鼠标显示抗实验者。拒绝，其中只有一个前爪或后肢中使用，并且其中鼠标拉过程中关措施。 </li><li>让鼠标拉格三次行，然后在笼子里返回它至少一分钟的休息期注:系列之间拉一个休息期是必要的鼠标恢复和避免习惯的形成。 </li><li>然后让鼠标执行四大系列拉，每个后跟一个短的休息时间。在这种方式的按钮鼠E有拉共15倍（3再换×5倍= 15拉）。 </li><li>确定最大握力和通过采取三个最高值的平均值出收集的15个值的归一化供体的重量。 </li><li>可选:通过计算前两个的平均值，最后两个系列拉1 +2 +3 = A，4 +5 +6 = B，10 +11 +12 = C和13 +14 +之间的递减确定疲劳15 = D。下式:（C + D）/（A + B）给出的1对小鼠不属于疲劳值。这可以用百分比来表示，使得鼠标不疲劳为0％的值和一个鼠标的前肢完全疲劳时具有100％的值。 </li></ol> 2。挂测试带悬挂试验，平衡，协调和肌肉状况进行评估。这些测试是基于小鼠都渴望继续挂在电线或网格，直到耗尽16的知识。有其中两个独特的悬挂试验在试验开始任只有两个前肢或四肢分别使用用金属丝或网格。使用线和网格挂试验是适于从最长的中止时间的方法<a href="http://www.treat-nmd.eu/downloads/file/sops/dmd/MDX/DMD_M.2.1.004.pdf" target="_blank">DMD_M.2.1.004.pdf</a>和<a href="http://www.treat-nmd.eu/downloads/file/sops/dmd/MDX/DMD_M.2.1.005.pdf" target="_blank">DMD_M.2.1.005.pdf</a>分别。用600秒的固定悬挂极限。大多数野生型小鼠的可以挂600秒，而营养不良的小鼠不能。以减少花费的时间进行该试验中，最大吊时间被设置到位。在此之前该脱落的线或网格小鼠给予长达两个多尝试。这样做是为了再保险，小鼠确实不能挂，不因笨拙下降。 <ol><li>吊试验与两肢<ol><li>设备设置:紧紧固定为2毫米厚的金属衣架用胶带架子和周围37保持架0;厘米上面有一层铺垫。注:另外，还可以使用55厘米，宽2mm厚的金属丝被紧紧地2纵看台之间的抵押。 37厘米的距离足以鼓励老鼠留挂，但也足够低，跌倒时防止老鼠从受伤。导线不应产生振动或在试验过程中置换出来，因为这可能会干扰小鼠的表现。 </li><li>通过尾部处理鼠标并把它的线附近。 </li><li>让鼠标抓住电线，只有两个前爪，及降低后肢在这样一种方式，鼠标只挂起与在电线上（ 图2B），这两个前爪。 </li><li>直接启动计时器，当鼠标被释放。释放后，强烈的小鼠试图抓住电线与所有的四肢和尾巴，这是允许的（ 图2C）。 </li><li>当鼠标显示了不恰当的行为（如平衡或故意跳下丝为SH拥有图2D和2E），直接通过更换鼠标的电线不停止计时器解决这个问题。 </li><li>当鼠标脱落的电线，停止计时并记录挂时间。 </li><li>当老鼠是能挂600秒，把他们赶走了电线，并将其返回到笼子里。在此之前限制落在老鼠都获得了最大两个的尝试。 </li><li>记录最大悬挂时间（ 即最长的试验），并使用它进行进一步的分析。 </li></ol></li><li>吊试验与四肢<ol><li>设备设置:使用一个手工制作的方形或一个大笼子里的老鼠或兔子本次测试的盖子。定位格25厘米以上柔软的被褥，以防止老鼠从坠落后伤害自己，而且要阻止小鼠故意跳下格。紧紧固定网格，使实验者不具备实验，这些运动过程中用手把握网格可能我nterfere用鼠标的性能。 </li><li>将鼠标放置在网格上，使其掌握它与它的四只爪子。 </li><li>颠倒的网格，使鼠标挂，直接启动定时器。 </li><li>测试会话结束的小鼠能够挂起600秒的持续时间。给予该脱落电网较早最多两个多尝试的小鼠。 </li><li>使用的最大吊时间（ 即 ，时间最长的试验）进行进一步分析。 </li></ol></li></ol> 3。旋转杆运行随着旋转试验的肌肉力量，协调，平衡和条件可确定17。 <ol><li>设备设置:对于此测试，小鼠必须在一个旋转管运行。确保稳定的速度设定为每分钟转速（rpm）5旋转，而当开始从5-45转在第15秒的速度增加。在此之后它具有保持其速度。 </li><li>将小鼠在转棒的管当它旋转时5rpm的缓慢稳定的速度。五只小鼠可同时进行测试。 </li><li>一旦开始所有小鼠被放置在运行。内的前15秒的管的速度加速从5-45转之后，它保持该速度。 </li><li>监视运行。运行时间由软件连续记录。运行时会自动停止，当鼠标脱落的管，因为这会激活位于管下方的时间条。重新定位该扭转的管朝向相反的方向，而不停止管旋转运行时，小鼠。 </li><li>结束测试阶段的小鼠都能够运行500秒的持续时间。给小鼠最多两次机会使他们能够改善他们的运行时间，当他们更早掉落。 </li><li>使用的最大运行时间（ 即最长的审判）进行进一步分析。 </li></ol> 4。跑步机练习该跑步机可以用三种方式中的预临床研究的工具。首先，强迫跑步机跑步可以作为本协议中所述加剧疾病的病理（参见: <a href="http://www.treat-nmd.eu/downloads/file/sops/dmd/MDX/DMD_M.2.1.001.pdf" target="_blank">DMD_M2.1.001.pdf</a> ）。其次，小鼠和治疗效果的在此的最大运转能力进行评估（参见让老鼠运行直到用尽该方法<a href="http://www.treat-nmd.eu/downloads/file/sops/dmd/MDX/DMD_M.2.1.003.pdf">DMD_M.2.1.003.pdf</a> ）。最后，跑步机的运行可使用之前的另一个功能测试用尽鼠标，这样它在所述第二测试13执行较差。这是通过如下文所述，之后直接跟随，任一个在协议1-3所描述的功能性测试的锻炼小鼠两次或每周3次进行。 <ol><li>设备设置:上有哪几个老鼠可以茹市售几个跑步机Ñ​​同时且为可抬高，持续时间和速度调整。一些跑步机配有网格提供低强度冲击，以鼓励老鼠跑。然而，mdx小鼠是对应力敏感的，并且可以很容易地通过一个温和的推带的运行方向上的手受到激励以友好的方式。因此，强烈建议不要使用冲击电网。一般来说，刺激手第一次运行会话期间，才需要。 </li><li>将小鼠在水平的跑步机。 </li><li>开始跑步机以12米/分的运行速度。较低的速度（8米/分）在老年小鼠中使用（&gt; 15个月），其中较高的速度容易导致疲劳。 </li><li>在第一次会议上，鼓励老鼠轻轻推他们，当他们靠近皮带的运行结束。 </li><li>当老鼠已经跑了30分钟的时间，将它们放回笼子。 </li><li>重复此每周两次， 例如 12周。 </LI&gt; <li>允许在需要的时候休息时间。例如，一些mdx小鼠必须停止运行，并应允许休息几个分钟。如果发生这种情况，把皮带脱落，给所有小鼠两分钟的休息期，把皮带上两分钟，在4米/分钟。在此之后，增加速度以12米/分钟，并允许小鼠以完成该协议。所有小鼠完成整个运行的协议是很重要的。 注意:如果mdx小鼠需要休息期间，考虑一个热身的30分钟运动方案之前。这种热身阶段组成:一个2分钟适应期在4米/分钟的速度，紧跟着的一个8分钟的热身以8米/分钟。 在我们的手中4-16周龄雌性mdx小鼠都能够完成30分钟运动方案没有休息。也有人报道，在年龄匹配的雄性mdx小鼠的老鼠45％的人需要休息期间，完成练习。热身协议减少停止12 s的量。 </li></ol>

<ol>
	<li>Blake, D. J., Weir, A., Newey, S. E., Davies, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Function+and+genetics+of+dystrophin+and+dystrophin-related+proteins+in+muscle.">Function and genetics of dystrophin and dystrophin-related proteins in muscle.</a> Physiol. Rev. 82, 291-329 (2002).</li><li>Hoffman, E. P., Brown, R. H., Kunkel, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dystrophin:+the+protein+product+of+the+Duchenne+muscular+dystrophy+locus.">Dystrophin: the protein product of the Duchenne muscular dystrophy locus.</a> Cell. 51, 919-928 (1987).</li><li>Bushby, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis+and+management+of+Duchenne+muscular+dystrophy,+part+1:+diagnosis,+and+pharmacological+and+psychosocial+management.">Diagnosis and management of Duchenne muscular dystrophy, part 1: diagnosis, and pharmacological and psychosocial management.</a> Lancet Neurol. 9, 77-93 .</li><li>Bulfield, G., Siller, W. G., Wight, P. A., Moore, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=X+chromosome-linked+muscular+dystrophy+(mdx)+in+the+mouse.">X chromosome-linked muscular dystrophy (mdx) in the mouse.</a> Proc. Natl. Acad. Sci U.S.A. 81, 1189-1192 (1984).</li><li>Bowles, D. 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=Phase+1+gene+therapy+for+Duchenne+muscular+dystrophy+using+a+translational+optimized+AAV+vector.">Phase 1 gene therapy for Duchenne muscular dystrophy using a translational optimized AAV vector.</a> Mol. Ther. 20, 443-455 (2012).</li><li>Cirak, 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=Exon+skipping+and+dystrophin+restoration+in+patients+with+Duchenne+muscular+dystrophy+after+systemic+phosphorodiamidate+morpholino+oligomer+treatment:+an+open-label,+phase+2,+dose-escalation+study.">Exon skipping and dystrophin restoration in patients with Duchenne muscular dystrophy after systemic phosphorodiamidate morpholino oligomer treatment: an open-label, phase 2, dose-escalation study.</a> Lancet. 378, 595-605 .</li><li>Goemans, N. 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=Systemic+administration+of+PRO051+in+Duchenne's+muscular+dystrophy.">Systemic administration of PRO051 in Duchenne's muscular dystrophy.</a> N. Engl. J. Med. 364, 1513-1522 (2011).</li><li>Malik, V., 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=Gentamicin-induced+readthrough+of+stop+codons+in+Duchenne+muscular+dystrophy.">Gentamicin-induced readthrough of stop codons in Duchenne muscular dystrophy.</a> Ann. Neurol. 67, 771-780 (2010).</li><li>Skuk, D., 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=First+test+of+a+&amp;#34;high-density+injection&amp;#34;+protocol+for+myogenic+cell+transplantation+throughout+large+volumes+of+muscles+in+a+Duchenne+muscular+dystrophy+patient:+eighteen+months+follow-up.">First test of a &amp;#34;high-density injection&amp;#34; protocol for myogenic cell transplantation throughout large volumes of muscles in a Duchenne muscular dystrophy patient: eighteen months follow-up.</a> Neuromuscul. Disord. 17, 38-46 (2007).</li><li>van Putten, 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=A+3+months+mild+functional+test+regime+does+not+affect+disease+parameters+in+young+mdx+mice.">A 3 months mild functional test regime does not affect disease parameters in young mdx mice.</a> Neuromuscul. Disord. 20, 273-280 (2010).</li><li>De Luca, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gentamicin+treatment+in+exercised+mdx+mice:+Identification+of+dystrophin-sensitive+pathways+and+evaluation+of+efficacy+in+work-loaded+dystrophic+muscle.">Gentamicin treatment in exercised mdx mice: Identification of dystrophin-sensitive pathways and evaluation of efficacy in work-loaded dystrophic muscle.</a> Neurobiol. Dis. 32, 243-253 (2008).</li><li>Radley-Crabb, H., 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=A+single+30min+treadmill+exercise+session+is+suitable+for+'proof-of+concept+studies'+in+adult+mdx+mice:+A+comparison+of+the+early+consequences+of+two+different+treadmill+protocols.">A single 30min treadmill exercise session is suitable for 'proof-of concept studies' in adult mdx mice: A comparison of the early consequences of two different treadmill protocols.</a> Neuromuscul. Disord. , (2011).</li><li>van Putten, 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+effects+of+low+levels+of+dystrophin+on+mouse+muscle+function+and+pathology.">The effects of low levels of dystrophin on mouse muscle function and pathology.</a> PLoS.One. , (2012).</li><li>Willmann, 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=Enhancing+translation:+Guidelines+for+standard+pre-clinical+experiments+in+mdx+mice.">Enhancing translation: Guidelines for standard pre-clinical experiments in mdx mice.</a> Neuromuscul. Disord. 1, 43-49 (2011).</li><li>Connolly, A. M., Keeling, R. M., Mehta, S., Pestronk, A., Sanes, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three+mouse+models+of+muscular+dystrophy:+the+natural+history+of+strength+and+fatigue+in+dystrophin-,+dystrophin/utrophin-,+and+laminin+alpha2-deficient+mice.">Three mouse models of muscular dystrophy: the natural history of strength and fatigue in dystrophin-, dystrophin/utrophin-, and laminin alpha2-deficient mice.</a> Neuromuscul. Disord. 11, 703-712 (2001).</li><li>Rafael, J. A., Nitta, Y., Peters, J., Davies, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testing+of+SHIRPA,+a+mouse+phenotypic+assessment+protocol+on+Dmd(mdx)+and+Dmd(mdx3cv)+dystrophin-deficient+mice.">Testing of SHIRPA, a mouse phenotypic assessment protocol on Dmd(mdx) and Dmd(mdx3cv) dystrophin-deficient mice.</a> Mamm. Genome. 11, 725-728 (2000).</li><li>Chapillon, P., Lalonde, R., Jones, N., Caston, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+development+of+synchronized+walking+on+the+rotorod+in+rats.+Effects+of+training+and+handling.">Early development of synchronized walking on the rotorod in rats. Effects of training and handling.</a> Behav. Brain Res. 93, 77-81 (1998).</li><li>Massett, M. P., Berk, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strain-dependent+differences+in+responses+to+exercise+training+in+inbred+and+hybrid+mice.">Strain-dependent differences in responses to exercise training in inbred and hybrid mice.</a> Am. J. Physiol. Regul. Integr. Comp. Physiol. 288, 1006-1013 (2005).</li><li>Lerman, I., 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=Genetic+variability+in+forced+and+voluntary+endurance+exercise+performance+in+seven+inbred+mouse+strains.">Genetic variability in forced and voluntary endurance exercise performance in seven inbred mouse strains.</a> J. Appl. Physiol. 92, 2245-2255 (2002).</li><li>Sharp, P. S., Jee, H., Wells, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiological+characterization+of+muscle+strength+with+variable+levels+of+dystrophin+restoration+in+mdx+mice+following+local+antisense+therapy.">Physiological characterization of muscle strength with variable levels of dystrophin restoration in mdx mice following local antisense therapy.</a> Mol. Ther. 19, 165-171 (2011).</li><li>Klein, S. 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=Noninvasive+in+vivo+assessment+of+muscle+impairment+in+the+mdx+mouse+model--a+comparison+of+two+common+wire+hanging+methods+with+two+different+results.">Noninvasive in vivo assessment of muscle impairment in the mdx mouse model--a comparison of two common wire hanging methods with two different results.</a> J. Neurosci. Methods. 203, 292-297 (2012).</li></ol>

作者什么都没有透露。

这里介绍的功能测试是可重复的，易于实施，并适用于野生型和独立的年龄的营养不良小鼠。该测试提供有用的工具，临床前评估肌肉功能，强度，条件和协调。当测试的化合物对疾病的自然史的影响，非侵入性的试验方法如下所述（前肢握力，这两个挂试验和旋转试验）可以很好地结合在这些测试是在连续数天进行功能测试制度。这些协议是不损害mdx小鼠，并且可以在纵向方式10...

假肥大型肌营养不良症（DMD）是最常见的神经肌肉疾病影响1:5,000新生男婴。这个严重的和渐进性肌肉萎缩病是由在DMD基因的突变，破坏开放读码框，并防止功能性肌养蛋白的合成所引起。肌纤维缺乏抗肌萎缩蛋白很容易受到运动性损伤。因肌肉的再生能力耗尽，并且由于受损的肌肉的慢性炎症，纤维由结缔组织和脂肪替代，随​​后导致功能丧失。一般情况下，DMD患者失去在第二个十年早期下肢行走。后来，又在胳膊和肩胛带肌肉受到影响，患者往往发展胸腰段脊柱侧凸，由于配套的脊髓肌肉不对称弱化。辅助通气，通常需要在晚十几岁或二十出头。呼吸和心脏衰竭铅死在第三或第四个十年1。 虽然致病基因已被发现在25年前2，没有可用于DMD的治疗。然而，改善医疗保健和使用糖皮质激素已在西方世界3提高寿命。通过使用像mdx小鼠的动物模型，主要步骤迈进的潜在治疗策略的发现已经作出。 mdx小鼠是最常用的DMD小鼠模型。它在小鼠DMD基因的外显子23点突变，因而缺乏抗肌萎缩蛋白4。在过去的几年中，许多建议策略已进展到临床试验5-9。在这些试验中，改善肌肉功能是主要终点，相关测试期间的前临床阶段测试化合物对肌肉功能的好处在小鼠中的重要性。 像DMD患者，也mdx小鼠的抗肌萎缩蛋白阴性肌纤维很容易受到运动损伤和比较C57BL/10ScSnJ野生型小鼠的肌肉功能受损。这种损害可以具有多种功​​能性测试进行评估。这些测试是无创，不与肌肉病理（ 如前肢握力，挂测试和转棒运行）干扰。因此它们可用于监测疾病的自然史或确定化合物对疾病进展的影响。得到一个在化合物对mdx小鼠的肌肉功能的影响的深度图像，功能测试制度不与疾病进展，包括所有这些测试的干扰可以用于10。 可替换地，跑步机强制运行可用于故意加重疾病的发展和测试化合物11的保护能力。跑步机也可以是在运行时间，直到耗尽测量12，或作为一种工具，疲劳mdx小鼠，使它们在随后的功能测试，确保在性能上治疗组13之间较大的差异表现稍逊作为观察指标。当选择功能测试，他们对疾病进展的影响应该测试营养不良小鼠像mdx小鼠14特别是当牢记。 我们在这里详细描述了如何执行基于来自TREAT-NMD网络中可用的标准作业程序以可靠和可重复的方式是最常用的功能测试。 <a href="http://www.treat-nmd.eu/resources/research-resources/dmd-sops/" target="_blank">点击这里访问TREAT-NMD</a> 。

<table border="0" cellpadding="0" cellspacing="0" width="832">
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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Mouse grip strength meter</td>
			<td style="width:331px;">Chatillon DFE (re-sold by Columbus Instruments)</td>
			<td style="width:119px;"># 80529</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hanging wire 2 limbs device</td>
			<td style="width:331px;">Cloth hanger or custom made device</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hanging wire 4 limbs device</td>
			<td style="width:331px;">Lid of rat cage or custom made device</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Rotarod</td>
			<td style="width:331px;">Ugo Basil</td>
			<td style="width:119px;"># 47600</td>
			<td></td>
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			<td height="21" style="height:21px;width:216px;">Treadmill for mice Exer 3/6</td>
			<td style="width:331px;">Columbus Instruments</td>
			<td style="width:119px;"># 1055SRM</td>
			<td></td>
		</tr>
	</tbody>
</table>

assessing functional performance in the mdx mouse model

假肥大型肌营养不良症（DMD）是一种严重和渐进性肌肉萎缩病也无法根治可用。然而，一些潜在的药物化合物和基因治疗方法已经进展到临床试验。随着改善肌肉功能是在这些试验中最重要的终点，很多重点放在了建立可靠，重现性好，易于进行功能测试，临床前评估肌肉功能，强度，条件，并在协调mdx小鼠模型的DMD。既侵袭性和非侵袭性测试可用。测试不恶化的疾病，可以用于确定疾病的自然史和治疗性干预（ 例如 ，前肢握力测试中，使用一个金属丝或栅和旋转杆运行两个不同的悬挂的测试）的影响。可替换地，跑步机强制运行可用于提高疾病进展和/或评估对疾病的病理治疗干预的保护作用。在这里，我们描述了如何在一个可靠和可重复的方式执行这些最常用的功能测试。使用这些协议基于标准作业程序使不同实验室之间的数据比较。

4-12周龄的野生型和mdx小鼠的前肢握力增加，再次降低了在老年小鼠。在力障碍已经可以在年轻的mdx小鼠进行观察。 9周龄的雌性小鼠的代表性数据示于图1A和 1B。虽然疲劳不菌株之间在这个年龄段有所不同的是，mdx小鼠比野生型小鼠弱。我们没有数据，但对疲劳性在旧的mdx和野生型小鼠。 获得可靠的和可重复的结果，多个评估需要由同...

Watch this Scientific Journal Video about Assessing Functional Performance in the Mdx Mouse Model at JoVE.com

Assessing Functional Performance in the Mdx Mouse Model

在对神经肌肉疾病的临床试验的主要终点是普遍改善肌肉功能。因此，评估潜在的治疗化合物对肌肉性能在小鼠模型中的临床前的效果是非常重要的。我们在这里介绍几种功能测试，以解决这一问题。

评估在功能性能<em&gt; MDX</em&gt;小鼠模型

Duchenne muscular dystrophy (DMD) is a severe and progressive muscle wasting disorder for which no cure is available. Nevertheless, several potential ...

assessing-functional-performance-in-the-mdx-mouse-model

Research

JoVE Journal

Behavior

Assessing Functional Performance in the Mdx Mouse Model

32.1K 次观看.  Leiden University Medical Center. 在对神经肌肉疾病的临床试验的主要终点是普遍改善肌肉功能。因此，评估潜在的治疗化合物对肌肉性能在小鼠模型中的临床前的效果是非常重要的。我们在这里介绍几种功能测试，以解决这一问题。 

视频: Assessing Functional Performance in the Mdx Mouse Model

Simultaneous Scalp Electroencephalography (EEG), Electromyography (EMG), and Whole-body Segmental Inertial Recording for Multi-modal Neural Decoding

Recent studies support the involvement of supraspinal networks in control of bipedal human walking. Part of this evidence encompasses studies, including our previous work, demonstrating that gait kinematics and limb coordination during treadmill walking can be inferred from the scalp electroencephalogram (EEG) with reasonably high decoding accuracies. These results provide impetus for development of non-invasive brain-machine-interface (BMI) systems for use in restoration and/or augmentation of gait- a primary goal of rehabilitation research. To date, studies examining EEG decoding of activity during gait have been limited to treadmill walking in a controlled environment. However, to be practically viable a BMI system must be applicable for use in everyday locomotor tasks such as over ground walking and turning. Here, we present a novel protocol for non-invasive collection of brain activity (EEG), muscle activity (electromyography (EMG)), and whole-body kinematic data (head, torso, and limb trajectories) during both treadmill and over ground walking tasks. By collecting these data in the uncontrolled environment insight can be gained regarding the feasibility of decoding unconstrained gait and surface EMG from scalp EEG.

Simultaneous Scalp Electroencephalography EEG, Electromyography EMG, and Whole-body Segmental Inertial Recording for Multi-modal Neural Decoding

Development of an effective brain-machine-interface (BMI) system for restoration and rehabilitation of bipedal locomotion requires accurate decoding of user's intent. Here we present a novel experimental protocol and data collection technique for simultaneous non-invasive acquisition of neural activity, muscle activity, and whole-body kinematics during various locomotion tasks and conditions.

同时头皮脑电图（EEG），肌电图（EMG）和全身分部惯性录制多模态神经解码

Recent  studies support the involvement of supraspinal networks in control of bipedal  human walking. Part of this evidence  encompasses studies, ...

科研

行为学

A Method for Investigating Age-related Differences in the Functional Connectivity of Cognitive Control Networks Associated with Dimensional Change Card Sort Performance

The ability to adjust behavior to sudden changes in the environment develops gradually in childhood and adolescence. For example, in the Dimensional Change Card Sort task, participants switch from sorting cards one way, such as shape, to sorting them a different way, such as color. Adjusting behavior in this way exacts a small performance cost, or switch cost, such that responses are typically slower and more error-prone on switch trials in which the sorting rule changes as compared to repeat trials in which the sorting rule remains the same. The ability to flexibly adjust behavior is often said to develop gradually, in part because behavioral costs such as switch costs typically decrease with increasing age. Why aspects of higher-order cognition, such as behavioral flexibility, develop so gradually remains an open question. One hypothesis is that these changes occur in association with functional changes in broad-scale cognitive control networks. On this view, complex mental operations, such as switching, involve rapid interactions between several distributed brain regions, including those that update and maintain task rules, re-orient attention, and select behaviors. With development, functional connections between these regions strengthen, leading to faster and more efficient switching operations. The current video describes a method of testing this hypothesis through the collection and multivariate analysis of fMRI data from participants of different ages.

This video presents a method of examining age-related changes in functional connectivity of cognitive control networks engaged by targeted tasks/processes. The technique is based on multi-variate analysis of fMRI data.

针对调查中的认知控制网络与关联维度变化卡片分类性能的功能连通性年龄相关性差异的方法

The ability to adjust behavior to sudden changes in the environment develops gradually in childhood and adolescence. For example, in the Dimensional ...

The Emotional Stroop Task: Assessing Cognitive Performance under Exposure to Emotional Content

The emotional Stroop effect (ESE) is the result of longer naming latencies to ink colors of emotion words than to ink colors of neutral words. The difference shows that people are affected by the emotional content conveyed by the carrier words even though they are irrelevant to the color-naming task at hand. The ESE has been widely deployed with patient populations, as well as with non-selected populations, because the emotion words can be selected to match the tested pathology. The ESE is a powerful tool, yet it is vulnerable to various threats to its validity. This report refers to potential sources of confounding and includes a modal experiment that provides the means to control for them. The most prevalent threat to the validity of existing ESE studies is sustained effects and habituation wrought about by repeated exposure to emotion stimuli. Consequently, the order of exposure to emotion and neutral stimuli is of utmost importance. We show that in the standard design, only one specific order produces the ESE.

The emotional Stroop effect (ESE) is the result of longer naming latencies to ink colors of emotion words than those of neutral words. This report refers to potential sources of confounding and includes a modal experiment that provides the means to control for them.

情绪Stroop任务:评估下暴露的认知表现的情感内容

The emotional Stroop effect (ESE) is the result of longer naming latencies to ink colors of emotion words than to ink colors of neutral words. The ...

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

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

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

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

鼠类精神运动警觉测试（rPVT）:一个在大鼠和小鼠的神经行为评估性能的方法

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

Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and validated for rat use. The Radial Water Tread (RWT) maze test of spatial memory (designed specifically for mice and requiring no swimming) has been shown previously to successfully distinguish between controlled cortical impact-induced TBI mice and sham controls. Here, a detailed protocol for this task is presented. The RWT maze capitalizes on the natural tendency of mice to avoid open areas in favor of hugging the sides of an apparatus (thigmotaxis). The walls of the maze are lined with nine escape holes placed above the floor of the apparatus, and mice are trained to use visual cues to locate the escape hole that leads out of the maze. The maze is filled with an inch of cold water, sufficient to motivate escape but not deep enough to require that the mouse swim. The acquisition period takes only four training days, with a test of memory retention on day five and a long-term memory test on day 12. The results reported here suggest that the RWT maze is a feasible alternative to rat-validated, swimming-based cognitive tests in the assessment of spatial memory deficits in mouse models of TBI.

Here we present a protocol for a mouse-specific test of cognition that does not require swimming. This test can be used to successfully distinguish controlled cortical impact-induced traumatic brain injury mice from sham controls.

使用径向水痕迷宫评估外伤性脑损伤小鼠模型中的空间记忆障碍

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and ...

Measuring Active and Passive Tameness Separately in Mice

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two behavioral components: a reluctance to avoid humans (passive tameness) and a motivation to approach humans (active tameness). To quantify these components in mice, we previously developed behavioral tests for active tameness, passive tameness, and the willingness to stay on a human hand, each designed to be completed within 3 min. The data obtained were used for selective breeding, with a large number of mice analyzed per generation. The active tameness test measures the movement of the mouse toward a human hand and the contact it engages in. The passive tameness test measures the duration of time that a mouse tolerates human touch. In the stay-on-hand test, a mouse is placed on a human hand and touched slowly using the thumb of that hand; the duration of time that the animal remains on the hand is measured. Here, we describe the test set-up and apparatus, explain the procedures, and discuss the appropriate data analysis. Finally, we explain how to interpret the results.

Tameness in animals includes a reduction of avoidance responses to humans (passive tameness) and an increase in active approaches to humans (active tameness). Here, we describe detailed protocols for three behavioral tests (active tameness, passive tameness, and stay-on-hand tests) to separately measure the active and passive tameness of mice.

小鼠主动和被动驯服的测定

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two ...

Biomechanical Analysis Methods to Assess Professional Badminton Players' Lunge Performance

Under the condition of simulating a badminton court in the laboratory, this study used the injury mechanism model to analyze the maximal right lunge movements of eight professional badminton players and eight amateur players. The purpose of this protocol is to study the differences in kinematics and joint moment of the right knee and ankle. A motion capture system and force plate were used to capture data of the joint movements of the lower extremity and the vertical ground reaction force (vGRF). Sixteen young men who did not have any sports injuries in the past 6 months took part in the study. The subjects performed a maximal right lunge from the start position with their right foot, stepping on and fully contacting with the force plate, hit the shuttlecock with an underhand stroke to the designated position in the backcourt, and then returned to the start/end position. All subjects wore the same badminton shoes to avoid a difference in impact from different badminton shoes. The amateur players showed a greater range of ankle movement and reverse joint moment on the frontal plane, and a larger internal joint rotation moment on the horizontal plane. The professional badminton players exhibited greater knee moment on the sagittal and frontal planes. Therefore, these factors should be considered in the development of the training program to reduce the risk of sports injuries in knee and ankle joints. This study simulates the real badminton court and calibrates the range of activities of each movement of the subjects so that the subjects complete the experimental action in a natural state with high quality. A limitation of this study is that it does not combine joint load and muscle activity. Another limitation is that the sample size is small and should be expanded in future studies. This research method can be applied to the lower limb biomechanical research of other footwork in the badminton project.

Here, we present a protocol to evaluate the differences in injury mechanisms between professional and amateur players when performing a badminton maximal right lunge movement by analyzing lower limb kinematics.

生物力学分析方法,以评估职业羽毛球运动员的隆起表现

Under the condition of simulating a badminton court in the laboratory, this study used the injury mechanism model to analyze the maximal right lunge ...

Assessing the Coherence of Parents' Short Narratives Regarding their Child Using the Five-Minute Speech Sample Procedure

Valid and efficient measures for assessing the quality of parent-child relationships are needed to facilitate research and evidence-based practice with parents. This paper focuses on such a method, namely Five-Minute Speech Sample-Coherence (FMSS-Coherence). In this method, a parent is asked to speak for five uninterrupted minutes about her/his child and their relationship. The resulted narrative is coded for coherence, namely the extent to which the parent provides&#160;in the narrative&#160;a clear, consistent, multidimensional and well-supported portrayal of the child. FMSS-Coherence is based on attachment research that shows that the coherence of parents&#8217; narratives is indicative of the quality of the parent-child relationship and child adjustment. It overcomes the limitations of attachment narrative measures which are typically labor intensive. FMSS-Coherence is less nuanced than extant attachment narrative measures. Yet, studies of families from different cultural backgrounds, across different child ages and in the context of typically developing children as well as children with special needs suggest that coherence can be reliably evaluated using the FMSS procedure. Furthermore, parents&#8217; FMSS-Coherence is associated with parenting quality and children&#8217;s socio-emotional adjustment. Thus, it holds promise for researchers and practitioners who seek a relatively time- and cost-effective method for assessing the coherence of parents&#8217; narratives regarding their child.

This paper introduces a method for assessing the coherence of parents' short narratives regarding their child and their relationship using the five-minute speech sample procedure.

使用五分钟言语样本程序评估家长对孩子的简短叙述的一致性

Valid and efficient measures for assessing the quality of parent-child relationships are needed to facilitate research and evidence-based practice with ...

A Task for Assessing the Impact of a Partner on the Speed and Accuracy of Motor Performance in Rats

To our knowledge, no study has examined the effect of mere presence on accuracy of performance in animals. Therefore, we developed an experimental task to measure rats&#8217; motor performance (speed and accuracy) in a social condition. Rats were trained to run on a runway and pull down a lever at the end of the runway. In testing, rats performed the task solitarily (single) or in the presence of a confederate rat beyond the lever (pair or a social condition). As indices of the performance speed, we measured the time needed to start running, run through the runway, and pull down the lever. As the index of performance accuracy, we counted the number of trials in which rats could pull down the lever during their first attempt. One-way and two-way repeated-measure analyses of variance were used to analyze the data. This run-and-pull task enabled us to examine the effect of the presence of another conspecific on both speed and accuracy of motor performance in one experiment. The results showed that rats performed the task faster but less accurately in pair sessions than in single sessions. This protocol would be a valid animal model to examine the effect of mere presence on speed and accuracy of motor performance in rats.

A procedure to measure the speed and accuracy of rats&rsquo; motor performance in a social condition is described. The protocol enables us to investigate the effect of the mere presence of others on speed and accuracy of motor performance in one experiment.

评估合作伙伴对大鼠电机性能速度和准确性的影响的任务

To our knowledge, no study has examined the effect of mere presence on accuracy of performance in animals. Therefore, we developed an experimental task to ...

Training Dogs for Awake, Unrestrained Functional Magnetic Resonance Imaging

We present a canine functional Magnetic Resonance Imaging (fMRI) training protocol that can be done in a cost-effective manner, with high-energy dogs, for acquisition of functional and structural data. This method of training dogs for awake, unrestrained fMRI employs a generalization procedure of stationing in several dissimilar locations to facilitate transfer of the stationing behavior to the real MRI scan environment; it does so without the need for extensive training time in the MRI scan environment, which can be expensive. Further, this method splits the training of a stationing (i.e., chin rest) behavior from desensitization to the MRI environment (i.e., 100+ decibel scan audio), the latter accomplished during dedicated Auditory Exposure conditioning sessions. The complete training and testing protocol required 14 hours and resulted in immediate transfer to novel locations. We also present examples of canine fMRI data that have been acquired from visual face processing and olfactory discrimination paradigms.

Magnetic resonance imaging (MRI) on unrestrained awake dogs is a new method with several advantages over imaging with physical or chemical restraint. This protocol introduces a cost-effective training method that minimizes training in the MRI environment, which can be expensive, and maximizes the subject pool available for canine functional MRI.

训练犬为觉醒，无节制的功能磁共振成像

We present a canine functional Magnetic Resonance Imaging (fMRI) training protocol that can be done in a cost-effective manner, with high-energy dogs, for ...