通过授予MF萨尔瓦多来自国家衰老研究所（AG040261）研究补助金的一部分提供资金用于这项工作。作者还要感谢维多利亚字段她的协助与工作相关的行为实验提出和Mackenzie果园，他协助建设逆止器的跑步机。

下面所有的程序都是按照机构动物照顾及使用委员会在路易斯安那州立大学健康科学中心 - 什里夫波特进行的，符合卫生指引研究所。 1，运动前程序注:所有在该强制运动养生相关手续的时间线图解如图1所示 。 <img alt="图1" fo:content-width="5in" src="/files/ftp_upload/51827/51827fig1highres.jpg" width="500" /> 图1:强制锻炼方案时间表 。这个时间表详细介绍每个星期的研究期间，这项工作方案的各个阶段的重要事件。预驯化阶段（周1-2）涉及实验者处理和基线运动活性。在适应阶段（​​3​​-4周），所有大鼠进行驯化培养3天瓦特在这里，他们坐在固定的跑步机和适应训练，跑步机，然后10天。在运动训练阶段（5-10周）是必需的最少两轮运动（1圆=连续12天），其次是2周的休息时间。行为和/或神经化学物质的测量可以在这些休息时间进行。此外，该方案可以被修改，以包括多轮的运动在这个阶段。体重是衡量整个研究过程中每周的各个阶段，在此之前（第一次会议）后（最后一届）这项工作方案的每一个阶段。跑步机运动训练成绩是1-4分配的所有锻炼和运动训练课程后，和射程，4为可能的最高得分。简单地说，对4训练成绩时，只无帮助锻炼整个训练会话分配。同样，指派3的训练成绩时，只需要最低限度的援助（定义为:协助小于25％的训练课从实验者的时间）。指定2的训练成绩时，只需要太多援助（定义为:协助培训会议的时间大于25％）的实验者在锻炼期间。最后，拨出1培训比分老鼠是不符合要求，未能完成练习会话。 <ol><li>抵达后的动物群体，独立的，单独容纳所有的科目为标准鞋盒笼。提供食物和水随意向受试者在整个研究。 </li><li>保持反12小时明暗周期（灯从1800到0600小时），在动物群体，并让受试者以适应这一周期至少一周。被检者的活动状态（暗）的周期中执行所有行为学实验，和1-2小时之前的明暗周期变化。 </li><li>之前的任何行为实验开始的两周内，办理所有考试科目数分茨每日熟习实验者的主体，并以降低的新颖性引起的应力不熟悉的环境中，如在运动室和跑步机。 </li><li>测量每个受试者的体重，每周一次，并立即前（第一次会议）后（最后一届）行使范式的每个主要区间。 </li></ol> 2，基线活动性评估:确定强制运动组分配<ol><li>评估为60分钟/天，每个主题的自发活动，连续5天，以建立基准的运动能力。开展自主活动会在自动旷场自发活动室包括一个有机玻璃箱（45×45×20 毫升 ），用红外光束的电网安装水平和垂直方向在产品说明书中建议的。 </li><li>在运动活性的评估，负载中的每一天的开始和设置在计算机和softwa重新计划连接到旷场运动活动室，以便每个会话期间，记录光束中断（自发活动）的数量。 </li><li>加入约2杯松木片的床上用品，以在每次会话的运动室的地板上。 </li><li>之前立即运出动物群体的自发活动室的主题在他们的家笼子里的自发活动的会话。加载对象到其指定的运动室下一个红灯，然后开始60分钟会话的计算机上一旦被放置到室内。 </li><li>在完成自发活动会话完成后，从运动室取出科目​​，并将其在红光下回到自己的家乡笼。立即返回受试者回到动物的殖民地。 </li><li>从室中取出被褥和每次会议后清洗室的墙壁和地板用50％的乙醇。 </li><li>每增加一个运动课程重复步骤2.3-2.6每天根据需要，直到所有的科目进行了检验。进行所有的运动活动会话在黑暗中，在动物的活性周期，和1-2小时之前的明暗周期变化。 </li><li>考虑到习惯化到活动室的可能性，旋转中的可用的腔室，使得自发活动是由前一日测量在不同的腔室的每个主题。 <ol><li>如果多个会话需要在一天内进行，变更测试，使得主题以不同的顺序，比前一交易日进行测试的顺序。 </li></ol></li><li>从由用于测量运动活动，确定与以下5个运动参数每只动物的基线运动活性的软件生成的数据:总行驶距离（厘米），水平活动（光束中断的次数），移动号码（开始动作的次数） ，花的时间移动（秒），并且移动速度（厘米/秒）。 <ol><li>为了分配练习的研究e和nonexercise组，计算出在所有5次的平均值为每上述步骤2.9中列出的5运动参数。 </li></ol></li><li>从这5个参数，确定锻炼和nonexercise（对照）组，使得同等范围内自发活动是代表在这两个群体。以这种方式，基于类似的运动功能配对大鼠在一起，并分配一个鼠至运动组和一个鼠的nonexercise基。因此，例如，配对的两只大鼠表现出最高平均所有5个运动参数的合计，并分配一个鼠至运动组和其他的nonexercise组（组分配是随机选择的）。其余的科目重复此过程。 <ol><li>保持配对整个强迫运动养生和对所有数据进行分析。 </li></ol></li><li>如果实验者有兴趣的运动对自发活动的影响，测量运动后的自发活动ü唱在步骤2.1-2.8.1上次跑步运动后开始一天中所述的相同步骤。注意:如果研究者有兴趣和/或自动自发活动的软件许可证，检验焦虑有关的措施（ 即在时间与外围活动室的中心略去）评估对象在自发活动的评估相对的焦虑。 </li></ol> 3，锻炼的跑步机<ol><li>进行在电动啮齿动物跑步机的所有平板运动有关的程序与由透明的有机玻璃墙中动物的活性周期分隔车道，以及1-2小时之前的明暗周期变化。 </li><li>代替位于每个车道的背面的电击线圈，用有机玻璃逆止专门设计，以适应在跑步机上，使得受试者可以连续运动而不与电休克线圈接触。为了防止逆止在培训班滑动，保持逆止到位，在C形夹1.5。 </li><li>由于实验者处理所需的跑步机运动训练，进行的灯还亮着所有的跑步机上锻炼会话。 </li><li>在他们的家笼子里的跑步机运输科前夕，跑步机运动训练。当跑步机是静止的，只加载到跑步机在其指定的车道通过向上滑动逆止刚好够，受试者可以逆止器下操纵。回向下滑动逆止到静止位置，一旦受到坐落在跑步机带。不要启动会话，直到所有的科目都被放置在其指定的车道为特定的会话。 <ol><li>在驯化的第一阶段，将每只大鼠的四车道的固定踏车的（如在步骤3.4中所述）为5-10分钟/天，连续3天中的一个。增加每个会话的时间每次连续ðAY（ 例如，5分钟，第1天，7分钟的第2天，并在第3天10分钟）作为大鼠熟悉跑步机装置，并减少对新颖性应力的潜力，在跑步机装置。重要提示:不要将老鼠在跑步机上（ 例如，停用的电击线圈），除了固定的皮带在这个阶段的任何地方。 </li></ol></li><li>在适应训练的第二阶段，将每只大鼠在固定跑步机的一个车道，在逆止门前。当所有的科目都到位，把跑步机上以缓慢的步行速度（ 例如，6米/分钟）。无论实验分组，培养所有大鼠在这个阶段上行走，跑步机，以确保从所述强制锻炼方案观察到的任何潜在的影响从运动训练导致，而不是简单地从锻炼的能力。 </li><li>由于跑步机的皮带开始滚动，确保所有的老鼠都走在了前进方向。协助大鼠为NEcessary由定向或轻轻捅了捅他们，直到他们正在向前推进。在适应训练期间，训练大鼠行走在跑步机上，使得它们不断地走在跑步机上未经实验者的援助。 <ol><li>通过逐渐增加速度过一段5-10分钟/天，连续10天适应各组大鼠的跑步机。以进一步促进遵守踏车运动，在所述第一5驯化会话训练大鼠在低速（6-8米/分钟）5-10分钟，并增加至适中的速度（9-10米/分钟）5在过去的5驯化会话-10分钟。 </li><li>确保所有的大鼠可被驯化训练期间的最后一届维持9-10米/分钟的速度进行10分钟。 </li></ol></li><li>在跑步机上锻炼会话完成时，如在步骤3.4中所述删除中用于加载对象的相同方式，跑步机主体。立即返回主题，以他们的家笼运回来的动物群体。 </li><li>擦拭每个单独的车道包括每个单独的踏车运动训练后的壁和跑步机带，用50％的乙醇。 </li><li>通过指定一个跑步锻炼的分数在每天的会议结束确定整个运动期间各组大鼠的运动能力。指定的4训练成绩时，只无帮助锻炼整个训练课。同样，指派3的训练成绩时，只需要最低限度的援助（定义为:协助培训会议的时间不超过25％）的实验者。指定2的训练成绩时，只需要太多援助（定义为:协助培训会议的时间大于25％）的实验者在锻炼期间。最后，拨出1培训比分老鼠是不符合要求，未能完成练习会话。 注:本活动计分制适用在某种程度上，其它研究19。 <ol><li>排除老鼠是无法完成驯化阶段，由于违规和/或防止主体行使（ 例如，这包括获得1跑步机运动训练成绩驯化训练的多个连续两天只）受伤。 </li></ol></li><li>平均在每个训练期间（ 如驯化培养和锻炼培训）在跑步机上的分数来确定每个主题的运动能力。确保所有的老鼠都能够赚到4跑步机上锻炼得分的适应期的最后一堂训练课中。 <ol><li>如果有的老鼠是不符合被他人在整个训练保持着顶级的运动速度，调整速度，这些老鼠，让他们能够继续行使的相同的时间量，但在整个培训课程以较慢的速度。在这种情况下，训练大鼠在组合模式慢吃了会议，并提高速度为主体适应较慢的速度。 </li></ol></li><li>为了避免对在跑步机上运动训练的跑步机车道偏移的可能性，旋转跑步机，让他们在训练中不同的车道，以后每次练习会话的可用通道中各组大鼠。 </li></ol> 4，跑步机运动训练<ol><li>这一天开始运动训练课程继去年驯化训练，并继续行使大鼠连续12天。在动物的活性周期进行这些会话和1-2小时之前的明暗周期变化。 </li><li>按照相同的设置程序，在驯化训练中使用的每个跑步机运动训练，如步骤3.2和3.4描述。 </li><li>在每届会议开始时，打开跑步机，以较低的，热身的速度（8米/分），比训练速度（9-11米/分钟），将维持throug豪特的演习训练的剩余部分。继续进行这场热身速度在会议开始时5-10分钟。调整速度9-11米/分钟的培训课程的其余部分。培训科目平均30分钟每一届整个12天运动训练。 </li><li>每次运动训练后，分配一个跑步机上锻炼的比分都使用在步骤3.9中描述的同样的运动评分系统只在运动组。不分配在跑步机上运动训练期间nonexercise鼠训练成绩。 </li><li>进行这项活动，分别nonexercise会议。将nonexercise大鼠在固定跑步机，他们的锻炼人群每届会议期间受过训练的时间相同。 </li><li>如步骤3.11所述，旋转每只大鼠的跑步机，让他们在训练中不同的车道，以后每次练习会话的可用通道之中。 </li><li>在完成一个循环完成运动，休息科目为14天。注意:根据从图2A和图2B中描述的试验性研究的结果是选择了该休息期。 </li><li>如果需要，进行各种行为和/或神经化学物质的评估，在12天的培训期结束。反之，则继续运动训练之后14天的休息时间随后的几轮。 </li></ol>

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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dichotomy+of+tyrosine+hydroxylase+and+dopamine+regulation+between+somatodendritic+and+terminal+field+areas+of+nigrostriatal+and+mesoaccumbens+pathways.">Dichotomy of tyrosine hydroxylase and dopamine regulation between somatodendritic and terminal field areas of nigrostriatal and mesoaccumbens pathways.</a> PLoS One. 7 (1), e29867 (2012).</li><li>Pruett, B. S., Salvatore, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nigral+GFR+&#945;-1+infusion+in+aged+rats+increases+locomotor+activity,+nigral+tyrosine+hydroxylase,+and+dopamine+content+in+synchronicity.">Nigral GFR &#945;-1 infusion in aged rats increases locomotor activity, nigral tyrosine hydroxylase, and dopamine content in synchronicity.</a> Mol. Neurobiol. 47 (3), 988-999 (2013).</li><li>Abercrombie, E. D., Zigmond, M. J., Bloom, ,. F. .. E., Kupfer, 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=Chapter+31:+Modification+of+central+categcholaminergic+systems+by+stress+and+injury:+Functional+significance+and+clinical+implications.">Chapter 31: Modification of central categcholaminergic systems by stress and injury: Functional significance and clinical implications.</a> Psychopharmacology: The Fourth Generation of Progress. 31, 355-361 (1995).</li><li>Ong, , 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=Neurobiological+consequences+of+acute+footshock+stress;+effects+on+tyrosine+hydroxylase+phosphorylation+and+activation+in+the+rat+brain+and+adrenal+medulla.">Neurobiological consequences of acute footshock stress; effects on tyrosine hydroxylase phosphorylation and activation in the rat brain and adrenal medulla.</a> J. Neurochem. , (2013).</li><li>Al-Jarrah, 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=Endurance+exercise+promotes+cardiorespiratory+rehabilitation+without+neurorestoration+in+the+chronic+mouse+model+of+parkinsonism+with+severe+neurodegeneration.">Endurance exercise promotes cardiorespiratory rehabilitation without neurorestoration in the chronic mouse model of parkinsonism with severe neurodegeneration.</a> J. Neurosci. 149 (1), 28-37 (2007).</li><li>Dishman, R. 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=Neurobiology+of+exercise.">Neurobiology of exercise.</a> Obesity. 14 (3), 345-356 (2006).</li></ol>

The authors declare no conflicts of interest.

有证据表明，生活方式的策略，如运动，可减少对慢性年龄有关的疾病和其主要症状，包括神经变性疾病如阿尔茨海默氏病和帕金森病20的风险。强制运动在跑步机上的好处，而不是自愿运动在跑轮，是研究者能够确定的频率，速度，和/或运动的强度和持续时间的练习会话。因此，具体的运动后的效果，如增加运动或认知能力，可以预测或预期下给定的方案的工作。此外，强迫跑步机锻?...

非侵入性的生活方式策略，可以防止或减轻损害，以认知和运动，两者常常与衰老有关，是获得牵引力为可行的做法，保持健康和福祉1,2。例如，相比与推进3个年龄相似的中年同行中到严格的训练，每周和一致的基础上的中年男人显著提高运动能力。此外，越来越多的证据表明，这些生活方式的策略，如锻炼，可以减轻或者甚至逆转与神经变性疾病如帕金森氏病（PD）4相关联的损伤。 努力了解的老化相关的损伤的分子机制，也已经朝向确定如何非侵入性策略，如锻炼，逆转或减轻，有助于这类损伤的机制。跑步机运动就是这样的一个STRAT埃及正在越来越多地应用在帕金森病5-7和认知功能障碍1款，其中仍在确定背后的改善运动或认知功能的机制。然而，它指出，老化是帕金森病的神经生物学背景是很重要的。因此，对于人类生存条件，以实现在动物模型中的任何运动利益的任何潜在的翻译必须考虑到老龄化的神经生物学背景。例如，在帕金森病的进展延缓运动障碍的代偿机制可以通过老化8的过程中受到损害。因此，按理说，锻炼范式必须开发，不仅考虑在任一的存在或不存在疾病的病理老化的影响，但也可以发起和维持老年啮齿类动物。 因此，考虑到大鼠品系上的神经生物学背景下老化，在选择应该由调查员进行精心考虑。一些大鼠品系可用于衰老研究，特别是菲舍尔344和布朗挪威/菲舍尔344传真:1（BNF）的混合体。常用只SD大鼠（远交株）也是适合于这种用途，因为它是常用的神经变性疾病模型，如6 - 羟基多巴胺PD模型。我们的实验室在衰老和神经退行性疾病的工作，同时使用只SD和BNF株。在这份报告中，我们将介绍使用结果两个菌株突出我们的工作协议。对于那些研究者严格聚焦于年龄相关的研究中，BNF应变提供了一些重要的优点。第一，它是比较不容易受到老化相关的疾病（如肿瘤），并具有出色的寿命（典型寿命大于30个月）相比于其它菌株。他们也有各种生理和行为的结果9，少变化d为也适合于探讨与衰老过程干预方法。此外，实验中如强迫运动需要相当的处理由研究者和BNF应变的温柔性格是有利的。纹状体和中脑组织多巴胺含量与老化相关的变化，以及自发活动的变化是BNF老鼠和灵长类动物10-11相似。我们的实验室还与表征和操纵纹状体和中脑多巴胺信号和自发活动的BNF应变12-16丰富的经验。因此，由于其他的神经生物学基础疾病的风险随着年龄增加，神经变性疾病的动物模型中应考虑使用啮齿类动物品系与使用在老化研究中的广泛记录。 我们的协议在此也解决了一些关键问题的调查必须考虑在他们的结果obtaine的解释d从演习的协议。啮齿动物跑步机设备（它的使用，我们将在本报告中突出显示），通常配有触电的线圈可以停用跑步机的每条车道的后面。然而，当这些电击线圈被激活时，一个小的电击递送给受试者，因为它来自于与线圈接触。这种策略通常被用在运动的研究，以促进遵守跑步锻炼。这是一个关键点的考虑，特别是对那些参与行为研究调查是由多巴胺和去甲肾上腺素的信号的影响。电气电击是一种生理应激，其在两个神经递质系统的影响是证据充分的，与酪氨酸羟化酶17-18的活性增加。因此，无论是增加神经递质的合成可能混淆的任何锻炼效果的诠释，使得研究者负责解释是否有任何在运动后的行为观察到的变化是由于严格的锻炼方案或电击的压力。重要的是，所提出的强制运动疗法不采用使用电击的在跑步机上锻炼的任何点或运动训练时段。 成功的运动疗法也需要遵守最高行使的考试科目。电击的，以实现符合性就业可能混淆实验结果的解释当从属措施涉及神经递质的信号是受电击的影响（如前面所讨论的）。因此，面临的挑战是让啮齿动物，尤其是鼠类岁，符合了运动处方。任何运动养生的理想目标是达到近100％达标，因为这将减少所需完成的运动养生动物的数量。具体而言，最大的合规性锻炼和运动养生的结果的解释可以直接使用作为几个运动前的程序，包括:1）反向明暗周期，使动物在其活动（唤醒）循环锻炼的结果在我们的运动处方得到E，2）确保运动性能基线是之前的隔离等控制和运动组，和3）将所述测试受试者的逐渐引入他们的锻炼方案的要求的适应期。在这里，上述实验的考虑，必须实现近100％达标措施，跑步机锻炼的老年啮齿类动物（&gt; 18月龄）进行评估并提交。最后，与运动相关联的环境中可能是有压力的，因此，本潜在混淆，以确定特定的行使的生理效应。我们的协议还包括在驯化阶段的每一个环节的nonexercise集团控制了暴露在跑步机上的潜在压力诱导的环境（包括MODEST跑步机上运行）和在跑步机上运动训练过程中把他们在固定的跑步机。因此，该协议的目的是描述需要单独确定运动的生理造成影响。

<table><tbody><tr><td>Exer-4 Treadmill for Rats</td><td>Columbus Instruments&amp;nbsp;</td><td>96157-2</td><td>Columbus has a variety of treadmills for animals with varying numbers of lanes and other options such as metabolic measurements</td></tr><tr><td>Animal Activity Meter: Opto-Varimex 4-Auto Track&amp;nbsp;</td><td>Columbus Instruments&amp;nbsp;</td><td>0170-R4</td><td>Columbus has several options for activity chambers with varying arena sizes, beam spacing, and sensors</td></tr></tbody></table>

getting to compliance in forced exercise in rodents: a critical standard to evaluate exercise impact in aging-related disorders and disease

有在运动对改善多种疾病有神经生物学基础的积极影响的认识大幅度增加;这些措施包括改善认知功能和物理性能。作为一个结果，存在增加在动物研究中使用的运动的数目。有人认为，强迫运动的一个内在价值是研究者拥有可影响运动对行为结果的影响因素的控制，尤其是运动频率的运动养生的，持续时间和强度。然而，遵从在强制运动疗法可能是一个问题，特别是如果是采用足部电击的潜在的困惑要避免。同样重要的是要考虑到，因为大多数的认知和运动障碍罢工老年个体，确定这些障碍应该考虑使用老年啮齿类动物与合规性的最高级别的运动的影响，以保证测试的苏最小需求bjects。在这里，相关的步骤和要达到近100％符合跑步机锻炼的老年啮齿类动物模型中考虑将介绍和讨论。尽管具体的锻炼方案所采用的调查，我们的协议应该是使用到调查是在强迫运动对衰老相关的损伤，包括老龄化相关的帕金森病和帕金森氏病的潜在影响，特别感兴趣。

在所有三个阶段都被迫锻炼方案的事件的时间线的示意图示于图1中 ，除了反转的光-暗循环，每天处理之前，任何实验测试，并驯化培养，其中受试者坐的第1阶段在固定的跑步机5-10分钟/天，连续3天，实施了以提高符合强制锻炼方案。这些程序被添加到协议中，以尽量减少与新颖的环境相关的应力，并且使受试者会更倾向于上行走的运动跑步机，如果他们已经熟悉的固定装置。这些?...

Watch this Scientific Journal Video about Getting to Compliance in Forced Exercise in Rodents: A Critical Standard to Evaluate Exercise Impact in Aging-related Disorders and Disease at JoVE.com

Getting to Compliance in Forced Exercise in Rodents: A Critical Standard to Evaluate Exercise Impact in Aging-related Disorders and Disease

Exercise may alleviate behavioral impairments associated with neurodegenerative diseases. Studies of the mechanisms underlying exercise-mediated effects have increased in disease models, but the neurobiological background accompanying these diseases, specifically aging, is likely not considered. Here, we illustrate measures to increase the likelihood of a successful exercise paradigm in aged rats.

让合规的强制锻炼鼠害:一项关键的标准在衰老相关的疾病和疾病评估运动的影响

There is a major increase in the awareness of the positive impact of exercise on improving several disease states with neurobiological basis; these ...

getting-to-compliance-forced-exercise-rodents-critical-standard-to

Universidad de Cartagena UDEC

Research

JoVE Journal

Behavior

14.1K 次观看.  Louisiana State University Health Sciences Center. Exercise may alleviate behavioral impairments associated with neurodegenerative diseases. Studies of the mechanisms underlying exercise-mediated effects have increased in disease models, but the neurobiological background accompanying these diseases, specifically aging, is likely not considered. Here, we illustrate measures to increase the likelihood of a successful exercise paradigm in aged rats.

视频: Getting to Compliance in Forced Exercise in Rodents: A Critical Standard to Evaluate Exercise Impact in Aging-related Disorders and Disease

Assessment of Murine Exercise Endurance Without the Use of a Shock Grid: An Alternative to Forced Exercise

Using laboratory mouse models, the molecular pathways responsible for the metabolic benefits of endurance exercise are beginning to be defined. The most common method for assessing exercise endurance in mice utilizes forced running on a motorized treadmill equipped with a shock grid. Animals who quit running are pushed by the moving treadmill belt onto a grid that delivers an electric foot shock; to escape the negative stimulus, the mice return to running on the belt. However, avoidance behavior and psychological stress due to use of a shock apparatus can interfere with quantitation of running endurance, as well as confound measurements of postexercise serum hormone and cytokine levels. Here, we demonstrate and validate a refined method to measure running endurance in na&#239;ve C57BL/6 laboratory mice on a motorized treadmill without utilizing a shock grid. When mice are preacclimated to the treadmill, they run voluntarily with gait speeds specific to each mouse. Use of the shock grid is replaced by gentle encouragement by a human operator using a tongue depressor, coupled with sensitivity to the voluntary willingness to run on the part of the mouse. Clear endpoints for quantifying running time-to-exhaustion for each mouse are defined and reflected in behavioral signs of exhaustion such as splayed posture and labored breathing. This method is a humane refinement which also decreases the confounding effects of stress on experimental parameters.

A method to assess exercise endurance in laboratory mice without the use of a shock grid is demonstrated. This method is a humane refinement that can decrease the confounding effects of stress on experimental parameters.

小鼠运动耐力的评估没有一个冲击电网的使用:另一种强迫运动

Using laboratory mouse models, the molecular pathways responsible for the metabolic benefits of endurance exercise are beginning to be defined. The most ...

科研

行为学

Supramaximal Intensity Hypoxic Exercise and Vascular Function Assessment in Mice

Exercise training is an important strategy for maintaining health and preventing many chronic diseases. It is the first line of treatment recommended by international guidelines for patients suffering from cardiovascular diseases, more specifically, lower extremity artery diseases, where the patients' walking capacity is considerably altered, affecting their quality of life. 
Traditionally, both low continuous exercise and interval training have been used. Recently, supramaximal training has also been shown to improve athletes' performances via vascular adaptations, amongst other mechanisms. The combination of this type of training with hypoxia could bring an additional and/or synergic effect, which could be of interest for certain pathologies. Here, we describe how to perform supramaximal intensity training sessions in hypoxia on healthy mice at 150% of their maximal speed, using a motorized treadmill and a hypoxic box. We also show how to dissect the mouse in order to retrieve organs of interest, particularly the pulmonary artery, the abdominal aorta, and the iliac artery. Finally, we show how to perform ex vivo vascular function assessment on the retrieved vessels, using isometric tension studies.

High-intensity training in hypoxia is a protocol that has been proven to induce vascular adaptations potentially beneficial in some patients and to improve athletes&#39; repeated sprint ability. Here, we test the feasibility of training mice using that protocol and identify those vascular adaptations using ex vivo vascular function assessment.

小鼠低血压运动的下位强度及血管功能评估

Exercise training is an important strategy for maintaining health and preventing many chronic diseases. It is the first line of treatment recommended by ...

Short Session High Intensity Interval Training and Treadmill Assessment in Aged Mice

High intensity interval training (HIIT) is emerging as a therapeutic approach to prevent, delay, or ameliorate frailty. In particular short session HIIT, with regimens less than or equal to 10 min is of particular interest as several human studies feature routines as short as a few minutes a couple times a week. However, there is a paucity of animal studies that model the impacts of short session HIIT. Here, we describe a methodology for an individually tailored and progressive short session HIIT regimen of 10 min given 3 days a week for aged mice using an inclined treadmill. Our methodology also includes protocols for treadmill assessment. Mice are initially acclimatized to the treadmill and then given baseline flat and uphill treadmill assessments. Exercise sessions begin with a 3 min warm-up, then three intervals of 1 min at a fast pace, followed by 1 min at an active recovery pace. Following these intervals, the mice are given a final segment that starts at the fast pace and accelerates for 1 min. The HIIT protocol is individually tailored as the speed and intensity for each mouse are determined based upon initial anaerobic assessment scores. Additionally, we detail the conditions for increasing or decreasing the intensity for individual mice depending on performance. Finally, intensity is increased for all mice every two weeks. We previously reported in this protocol enhanced physical performance in aged male mice and here show it also increases treadmill performance in aged female mice. Advantages of our protocol include low administration time (about 15 min per 6 mice, 3 days a week), strategy for individualizing for mice to better model prescribed exercise, and a modular design that allows for the addition or removal of the number and length of intervals to titrate exercise benefits.

Short session (&#8804;10 min) high intensity interval training (HIIT) is emerging as an alternative to longer exercise modalities, yet the shorter variants are rarely modeled in animal studies. Here, we describe a 10 min, 3 day a week, uphill treadmill HIIT protocol that enhances physical performance in male and female aged mice.

老年小鼠短期高强度间隔训练与跑步机评价

High intensity interval training (HIIT) is emerging as a therapeutic approach to prevent, delay, or ameliorate frailty. In particular short session HIIT, ...

A Method to Study the Impact of Chemically-induced Ovarian Failure on Exercise Capacity and Cardiac Adaptation in Mice

The risk of cardiovascular disease (CVD) increases in post-menopausal women, yet, the role of exercise, as a preventative measure for CVD risk in post-menopausal women has not been adequately studied. Accordingly, we investigated the impact of voluntary cage-wheel exercise and forced treadmill exercise on cardiac adaptation in menopausal mice. The most commonly used inducible model for mimicking menopause in women is the ovariectomized (OVX) rodent. However, the OVX model has a few dissimilarities from menopause in humans. In this study, we administered 4-vinylcyclohexene diepoxide (VCD) to female mice, which accelerates ovarian failure as an alternative menopause model to study the impact of exercise in menopausal mice. VCD selectively accelerates the loss of primary and primordial follicles resulting in an endocrine state that closely mimics the natural progression from pre- to peri- to post-menopause in humans. To determine the impact of exercise on exercise capacity and cardiac adaptation in VCD-treated female mice, two methods were used. First, we exposed a group of VCD-treated and untreated mice to a voluntary cage wheel. Second, we used forced treadmill exercise to determine exercise capacity in a separate group VCD-treated and untreated mice measured as a tolerance to exercise intensity and endurance.

Two exercise paradigms were tested on a newly developed chemically induced menopausal mouse model to examine the impact of menopause on exercise capacity and cardiac adaption to exercise.

一个方法研究的化学诱导卵巢早衰的运动能力和适应心脏的影响在小鼠

The risk of cardiovascular disease (CVD) increases in post-menopausal women, yet, the role of exercise, as a preventative measure for CVD risk in ...

医学

Tracking Individual Running Metrics in Mice Using a Voluntary Wheel Running Protocol that Minimizes Social Isolation

Voluntary wheel running (VWR) in rodents is used to evaluate performance and endurance and to simulate exercise. In contrast to other exercise paradigms that require aversive stimulus to induce active movement, wheel running is voluntary. VWR generally consists of placing a running wheel inside the mouse home cage. However, analyzing exercise parameters can typically only be conducted when mice are individually housed, while overall cage effects can be reported when mice are group-housed. Therefore, the current models force experimenters to choose between measuring individual exercise parameters in single-housed mice or measuring group effects in group-housed mice; depending on the research question and model, this could be limiting. 
Therefore, the goal of this method is to demonstrate a protocol that assesses VWR in a mouse model by quantifying running parameters in individual mice, with minimal isolation. We report on device design, preoperative preparations, and experiment setup. Results of a 6-week training protocol using this VWR model suggest it provides sufficient stimulus to increase spatial object recognition memory, comparable to the clinical effect of exercise. This simple and inexpensive model facilitates investigation into the effect of VWR on multiple measurements of wheel activity, minimizing factors that could limit or affect experimental outcomes.

This protocol describes an effective voluntary wheel-running model that can be implemented in various research contexts. Unlike other voluntary running models, this method avoids isolation housing while tracking running metrics in individual mice.

Voluntary wheel running (VWR) in rodents is used to evaluate performance and endurance and to simulate exercise. In contrast to other exercise paradigms ...

Conducting Maximal and Submaximal Endurance Exercise Testing to Measure Physiological and Biological Responses to Acute Exercise in Humans

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic responses to acute exercise are predominantly influenced by the duration and intensity of the exercise regimen. As exercise is increasingly thought of as a therapeutic treatment and/or diagnostic tool, it is important that standardizable methodologies be utilized to understand the variability and to increase the reproducibility of exercise outputs and measurements of responses to such regimens. To that end, we describe two different cycling exercise regimens that yield different physiologic outputs. In a maximal exercise test, exercise intensity is continually increased with a greater workload resulting in an increasing cardiopulmonary and metabolic response (heart rate, stroke volume, ventilation, oxygen consumption and carbon dioxide production). In contrast, during endurance exercise tests, the demand is increased from that at rest, but is raised to a fixed submaximal exercise intensity resulting in a cardiopulmonary and metabolic response that typically plateaus. Along with the protocols, we provide suggestions on measuring physiologic outputs that include, but are not limited to, heart rate, slow and forced vital capacity, gas exchange metrics, and blood pressure to enable the comparison of exercise outputs between studies. Biospecimens can then be sampled to assess cellular, protein, and/or gene expression responses. Overall, this approach can be easily adapted into both short- and long-term effects of two distinct exercise regimens.

To assess the influence of exercise intensity on physiologic and biologic responses, two different exercise testing protocols were utilized. Methods outlining exercise testing on a cycle ergometer as an incremental maximal oxygen consumption test and endurance, steady state submaximal endurance test are described.

进行最大和次极大耐力运动测试, 测量人类急性运动的生理和生物学反应

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic ...

Dosage-Adjusted Resistance Training in Mice with a Reduced Risk of Muscle Damage

Progressive resistance training (PRT), which involves performing muscle contractions against progressively greater external loads, can increase muscle mass and strength in healthy individuals and in patient populations. There is a need for precision rehabilitation tools to test the safety and effectiveness of PRT to maintain and/or restore muscle mass and strength in preclinical studies on small and large animal models. The PRT methodology and device described in this article can be used to perform dosage-adjusted resistance training (DART). The DART device can be used as a standalone dynamometer to objectively assess the concentric contractile torque generated by the ankle dorsiflexors in mice or can be added to a pre-existing isokinetic dynamometry system. The DART device can be fabricated with a standard 3D printer based on the instructions and open-source 3D print files provided in this work. The article also describes the workflow for a study to compare contraction-induced muscle damage caused by a single bout of DART to muscle damage caused by a comparable bout of isometric contractions (ISOM) in a mouse model of limb-girdle muscular dystrophy type 2B/R2 (BLAJ mice). The data from eight BLAJ mice (four animals for each condition) suggest that less than 10% of the tibialis anterior (TA) muscle was damaged from a single bout of DART or ISOM, with DART being less damaging than ISOM.

The present protocol describes a unique technique called dosage-adjusted resistance training (DART), which can be incorporated into precision rehabilitation studies performed in small animals, such as mice.

降低肌肉损伤风险的小鼠剂量调整阻力训练

Progressive resistance training (PRT), which involves performing muscle contractions against progressively greater external loads, can increase muscle ...

神经科学

Activating Autophagy by Aerobic Exercise in Mice

Autophagy is a lysosomal degradation pathway essential for cell homeostasis, function and differentiation. Under stress conditions, autophagy is induced and targets various cargos, such as bulk cytosol, damaged organelles and misfolded proteins, for degradation in lysosomes. Resulting nutrient molecules are recycled back to the cytosol for new protein synthesis and ATP production. Upregulation of autophagy has beneficial effects against the pathogenesis of many diseases, and pharmacological and physiological strategies to activate autophagy have been reported. Aerobic exercise is recently identified as an efficient autophagy inducer in multiple organs in mice, including muscle, liver, heart and brain. Here we show procedures to induce autophagy in vivo by either forced treadmill exercise or voluntary wheel running. We also demonstrate microscopic and biochemical methods to quantitatively analyze autophagy levels in mouse tissues, using the marker proteins LC3 and p62 that are transported to and degraded in lysosomes along with autophagosomes.

Autophagy activation is beneficial in the prevention of a number of diseases. One of the physiological approaches to induce autophagy in vivo is physical exercise. Here we show how to activate autophagy by aerobic exercise and measure autophagy levels in mice.

小鼠中自噬激活有氧运动

Autophagy is a lysosomal degradation pathway essential for cell homeostasis, function and differentiation. Under stress conditions, autophagy is induced ...

生物学

Versatility of Protocols for Resistance Training and Assessment Using Static and Dynamic Ladders in Animal Models

Resistance training is a physical exercise model with profound benefits for health throughout life. The use of resistance exercise animal models is a way to gain insight into the underlying molecular mechanisms that orchestrate these adaptations. The aim of this article is to describe exercise models and training protocols designed for strength training and evaluation of resistance in animal models and provide examples. In this article, strength training and resistance evaluation are based on ladder climbing activity, using static and dynamic ladders. These devices allow a variety of training models as well as provide precise control of the main variables which determine resistance exercise: volume, load, velocity, and frequency. Furthermore, unlike resistance exercise in humans, this is a forced exercise. Thus, aversive stimuli must be avoided in this intervention to preserve animal welfare. Prior to implementation, a detailed design is necessary, along with an acclimatization and learning period. Acclimatization to training devices, such as ladders, weights, and clinical tape, as well as to the manipulations required, is necessary to avoid exercise rejection and to minimize stress. At the same time, the animals are taught to climb up the ladder, not down, to the resting area on the top of the ladder. Resistance evaluation can characterize physical strength and permit adjusting and quantifying the training load and the response to training. Furthermore, different types of strength can be evaluated. Regarding training programs, with appropriate design and device use, they can be sufficiently versatile to modulate different types of strength. Furthermore, they should be flexible enough to be modified depending on the adaptive and behavioral response of the animals or the presence of injuries. In conclusion, resistance training and assessment using ladders and weights are versatile methods in animal research.

The present protocol describes resistance training and testing using static and dynamic ladders in animal models.

在动物模型中使用静态和动态梯子进行阻力训练和评估方案的多功能性

Resistance training is a physical exercise model with profound benefits for health throughout life. The use of resistance exercise animal models is a way ...

A Simple and Inexpensive Running Wheel Model for Progressive Resistance Training in Mice

Previously developed rodent resistance-based exercise models, including synergistic ablation, electrical stimulation, weighted-ladder climbing, and most recently, weighted-sled pulling, are highly effective at providing a hypertrophic stimulus to induce skeletal muscle adaptations. While these models have proven invaluable for skeletal muscle research, they are either invasive or involuntary and labor-intensive. Fortunately, many rodent strains voluntarily run long distances when given access to a running wheel. Loaded wheel running (LWR) models in rodents are capable of inducing adaptations commonly observed with resistance training in humans, such as increased muscle mass and fiber hypertrophy, as well as stimulation of muscle protein synthesis. However, the addition of moderate wheel load either fails to deter mice from running great distances, which is more reflective of an endurance/resistance training model, or the mice discontinue running nearly entirely due to the method of load application. Therefore, a novel high-load wheel running model (HLWR) has been developed for mice where external resistance is applied and progressively increased, enabling mice to continue running with much higher loads than previously utilized. Preliminary results from this novel HLWR model suggest it provides sufficient stimulus to induce hypertrophic adaptations over the 9 week training protocol. Herein, the specific procedures to execute this simple yet inexpensive progressive resistance-based exercise training model in mice are described.

This procedure describes a translatable progressive loaded running wheel resistance training model in mice. The primary advantage of this resistance training model is that it is entirely voluntary, thus reducing stress for the animals and the burden on the researcher.

一种简单且廉价的跑轮模型，用于小鼠的渐进式阻力训练

Previously developed rodent resistance-based exercise models, including synergistic ablation, electrical stimulation, weighted-ladder climbing, and most ...