The authors would like to thank Dr. Jhi-Joung Wang, who is the Vice Superintendent of Education at Chi-Mei Medical Center, and Dr. Chih-Chan Lin from the Laboratory Animal Center, Department of Medical Research, Chi-Mei Medical Center, 901 Zhonghua, Yongkang Dist., Tainan City 701, Taiwan, for providing the shooting venue. They would also like to thank Miss Ling-Yu Tang and Mr. Chung-Ham Wang from the Department of Medical Research, Chi-Mei Medical Center, Tainan, Taiwan, for their valuable assistance in demonstrating the prototype system in real experiments with rats. The author gratefully acknowledges the support provided for this study by the Ministry of Science and Technology (MOST 104- 2218-E-167-001-) of Taiwan.

伦理声明:实验过程是由科学技术实验动物中心南台科技大学，国家科学委员会，中华民国（台湾台南）的动物伦理委员会批准。 1.构建跑轮结构注:所有丙烯酸应该是透明的。用清水清洗拆卸车轮，然后用酒精擦拭每次使用后的橡胶履带和亚克力板材。 <ol><li>得到丙烯酸跑轮为55厘米，直径在宽15厘米 注意:此轮比传统传动轮（直径= 35厘米;宽度= 12cm）中较大的（F igure 1A）。 </li><li>使用切割机，切四分之一圆开口进入运行车轮的一侧充当一个入口和一个出口以及对大鼠（ 图1B）。将高摩擦橡胶履带上一层ACR的内ylic轮（ 图1B）。 </li><li>放置铁棒用轴承连接正在运行的车轮（ 图1B）。放置在运行车轮的两侧的两个丙烯酸三角形列充当支撑框架（ 图1B）。 </li><li>附加一个1mm厚的半圆形，透明压克力板用螺钉两个三角柱的外部侧面。使用此表来部署红外传感器。确保丙烯酸系片材是从正在运行的轮的每一侧约3厘米。 </li></ol> 2.部署红外传感器和定义的有效行使区注意:考虑到正在运行的车轮尺寸和在红外系统的设计大鼠长度。一只老鼠只能触发一次一个传感器。在该实验中，老鼠是厘米20和23之间长。 <ol><li>钻一个洞，压克力板材每45°（ARC间隔= 21厘米），随着距离两孔之间是大致相当于一个测试大鼠的长度。使孔的大小相同的红外传感器（ 图2A）。 注:对于传统MRWs，钻一个孔每隔70°（ARC间隔= 21厘米， 图2B）。 </li><li>期间PRW实验，维持大鼠在0℃之间运行，以135℃的稳定状态。 注:这样定义这个区域作为有效演习区域，同时鉴于所有其他部分为无效运动领域。对于传统MRWs，限定有效行使区域为140°（ 图2B）0°之间的部分。 </li></ol> 3.驾驶跑轮<ol><li>使用无刷DC电动机和电动机驱动器来驱动的运行车轮。 </li><li>安装在电机的中轴线（ 图1B）一个直径10厘米的橡胶盘。 </li><li>使用铁框和弹簧以支撑电机，从连接橡胶磁盘电机的中心轴线到正在运行的车轮的外侧。 注:弹簧必须用螺丝合作，允许动态电机高度调节，防止橡胶盘被断开，因为弹簧松动的运行轮距。 </li><li>操作马达来驱动使用微控制器10厘米直径的橡胶盘，并观察车轮转动由于橡胶盘和车轮的跑道之间的摩擦，形成一个机动运行车轮的平台。 </li><li>安装4红外传感器顺序地为0°至135°（ 图2A）之间。 注:对于传统MRWs，0°之间安装传感器以140°（ 图2B）。 </li><li>连接4双安装在两个丙烯酸类片材，使用单芯电缆，从而形成一定位跑轮系统微控制器的一般销红外线传感器。 </li></ol> 4.构建自适应Accelerati曲线上<ol><li>官方3周运动训练开始前三天，通过手动操作运转轮系老鼠。 注意:我们的目标是让大鼠熟悉的运行环境，并且是检测各大鼠是否能够忍受在20米/分钟运行。 <ol><li>在手工操作的培训，逐步加快运行速度，直到老鼠是无法跟上时代的步伐。发生这种情况时，降低速度，直到老鼠重新获得稳定的运行速度，然后逐步直到老鼠达到20米/分钟（ 图3虚线）再次提高速度。该手册的培训涉及7只构建培训曲线。 </li></ol></li><li>使用数字式，以适应在手动测试的第3天的测定数据，计算最靠近手动训练（有圆圈的曲线， 图3）的加速度曲线。拟合方程1的原始数据，其中C INI = 8，C 翅 </suB&gt; = 20，τ= 30表示的初始速度，最后的速度和时间常数，分别地）。 注:此公式适应老鼠的身体状况。因此，请参照所计算的曲线作为自适应加速训练模式。 <img alt="式（1）" src="/files/ftp_upload/54354/54354eq1.jpg" /> （1） </li><li>用公式1为正式训练1周。 </li><li>对于周2和培训3，调整公式1的参数，也就是说，改变从12到22，让速度达到30米/分钟。 </li></ol> 5.控制软件程序注:独占开发代码为基于微控制器的马达的操作和用于从红外传感器用于随后的数据分析的计算机信号传输。 <ol><li>用C编程语言编写包含一个主程序和两个中断服务例程的定时器的软件控制程序微控制器18。 <ol><li>确保主程序初始化微控制器的寄存器和构造在微控制器的存储器中的自适应加速曲线模型。 </li><li>使用定时器0的中断服务程序激活自适应加速曲线，并计算出整个训练持续时间。 </li><li>用定时器1的中断服务例程，以从红外传感器提取信号数据和将数据传送到计算机。 </li><li>使用主节目录制0°的位置来调整运行车轮的速度。 </li></ol></li><li>一旦在0℃的接收红外传感器被触发，它解释为跌倒发生率，这是由主程序积累。此刻秋季发生率的发生次数击中大鼠位置检测数的10％的阈值时，自动deaccelerate运行车轮作为训练大鼠安全措施。注:速度O˚F运行车轮降低，直到老鼠可以返回到安全区域（0°到135），并保持稳定的运行状态的安全措施。 </li></ol> 6.操作定位跑轮系统<ol><li>打开微控制器和等待操作员按下按钮开始每星期的训练模式。 <ol><li>按"开始"按钮，开始培养模式1周。 注:电机，直到达到20米/分钟，自动后30分钟停止基于自适应加速曲线自动加速。 </li><li>按"开始"按钮，开始培养模式2周。 注:电机，直到达到30米/分钟，自动后30分钟停止基于自适应加速曲线自动加速。 </li><li>按"开始"按钮，开始培养模式为3周。 注:电机自动accelera基于自适应加速度曲线上，直到达到30米/分钟，并自动TES 60分钟后停止。 注意:在整个培养过程中，发送从红外传感器接收到的计算机无线地将信号数据。 </li></ol></li><li>使用计算机，分析所述位置数据，以获得在整个锻炼过程中有效的锻炼度量（ <img alt="公式（2）" src="/files/ftp_upload/54354/54354eq2.jpg" /> ）。见公式2。 <img alt="公式3" src="/files/ftp_upload/54354/54354eq3.jpg" /> （2） 注:EEE，EED和IED代表有效行使的措施，有效和无效的锻炼持续时间分别。 </li></ol> 7.训练大鼠<ol><li>随机分为成年雄性SD大鼠分为五组（n = 9每个组）:假，控制，跑步机，MRW和PRW组。 </li><li>进行3周的运动训练三个运动组， 即 TReadmill，MRW和PRW群体，而假组和对照组则没有。 注意:每个练习组3周的运动训练为20米/分钟的第1周30分钟，30米/分钟30分钟2周期间和30米/分钟60分钟3周期间。 </li></ol> 8.动物和中风模型<ol><li>如第说明。 7.1，随机分为所有涉及的成年雄性SD大鼠，250〜280 g的体重，分成5组。 </li><li>权衡所有的动物，以确保准确的药物剂量计算。麻醉用戊巴比妥钠的大鼠（25毫克/千克，腹膜内[IP]）和含混合物氯胺酮（4.4毫克/千克，肌肉注射[IM]），阿托品（0.02633毫克/千克，[IM]）和赛拉嗪（6.77毫克/公斤，[IM]）。 <ol><li>通过监测呼吸率（正常70-115次/ min），节奏，呼吸，粘膜颜色和反射的定期检测的深度， 例如脚趾捏，尾捏，眼睑/睫毛评估麻醉深度和眼睑。 </li></ol></li><li>将温度探头插入直肠，并使用独立的加热灯保持37之间的直肠温度37.5℃。 </li><li>通过插入灯丝插入内颈动脉以阻塞通过外颈动脉的方法19中脑动脉的孔诱导局灶性缺血，暂时性大脑中动脉闭塞（MCAO）。 <ol><li>执行在假手术动物相同的操作程序，而不要插入灯丝插入颈内动脉。维持脑缺血1小时后，取出长丝，关闭切口，然后离开1厘米尼龙缝合突出，这可以被收回，以允许再灌注。 </li></ol></li><li>辖皮下（sc）止痛剂（丁丙诺啡（0.05毫克/公斤，SC））的注射剂，成动物镇痛一天两次3天。 </li></ol> 9.评估神经损伤<ol><li> EVAL审视你们神经和运动功能，分别由神经严重性评分（MNSS）20和一个倾斜面测试21。 注:MNSS是电动机（肌肉状态，异常的运动），感觉（视觉，触觉和本体）和反射测试的复合物。给予对未执行任务一点。在0-18的刻度率神经功能（正常得分= 0;最大缺陷评分= 18）。 </li><li>前一天和每日评估所有大鼠中的行为性能方面在手术后7天的时间跨度。 </li><li>测量使用斜面的大鼠后肢握力。 <ol><li>放置在每天的基础上的倾斜攀登装置大鼠和试验前1周使其恢复的大鼠的装置和试验条件。 </li><li>放置在该装置的每个大鼠和鼓励大鼠爬上平台直到该装置的过程中的环境适应期的顶部。 </li><li>放置在老鼠与在测试过程中向下方的头装置的顶部。确保开始在25°的角度大鼠停留的体轴沿一个20×20厘米2橡胶带肋表面上的倾斜面。 </li><li>增加角度动态使用与步进电机相连的滚珠丝杠，以确定在该动物可容纳于所述平面的最大角度。增大倾斜面的角度逐渐直到鼠标未能保持在倾斜面，然后检测一个往下滑事件。倾斜面的基线程度是在开始时25°。 </li><li>请两位观察员（不知道已经给大鼠什么待遇），独立审查和评分所有的行为测试， 即左侧和右侧的最大角度的平均值。 </li></ol></li><li> Sacri科幻CE所有第7天的动物缺血后。 （100毫克/千克，IP戊巴比妥钠）用盐水灌注22深麻醉下的动物的心</SUP&gt;。删除，然后浸入冷生理盐水的脑组织进行5分钟，然后将其切成使用组织切片机222.0毫米冠状切片。 </li><li>淹没新鲜脑切片成2，3，5-三苯基氯化四唑（TTC）中于37℃进行30分钟，然后在4℃转移片至5％的甲醛溶液为音响xation 24小时。将染色的脑片上有机玻璃支架上。 </li><li>拍摄TTC染色切片用使用链接到装载有图像处理软件的个人计算机的CCD照相机校准的刻度。使用半自动化图像分析系统，以及估算从每个TTC染色的脑切片23上的梗塞面积（mm 2）。 </li><li>计算所有脑片梗死区的总和，每一片总梗死体积。标记分别对2毫米厚的片每侧未染色区域（缺血性脑），然后计算梗死体积和的平均值。 </li><li> Calcu后期修正梗死体积（CIV）作为 CIV = {长线（RT-RI）} <img alt="公式4" src="/files/ftp_upload/54354/54354eq4.jpg" /> D（3） 注意:如果Lt和Rt表示为mm 2的左和右半球的区域，分别RI为以mm 2梗塞面积，d = 2毫米是切片厚度。 </li></ol>

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<li>Longa, E. Z., Weinstein, P. R., Carlson, S., Cummins, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Reversible+middle+cerebral+artery+occlusion+without+craniectomy+in+rats%5BTitle%5D)+AND+%22Stroke%22%5BJournal%5D)">Reversible middle cerebral artery occlusion without craniectomy in rats.</a> Stroke. 20 (1), 84-91 (1989).</li>
<li>Chen, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Therapeutic+benefit+of+intravenous+administration+of+bone+marrow+stromal+cells+after+cerebral+ischemia+in+rats%5BTitle%5D)+AND+%22Stroke%22%5BJournal%5D)">Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats.</a> Stroke. 32 (4), 1005-1011 (2001).</li>
<li>Chang, M. -W., Young, M. -S., Lin, M. -T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+inclined+plane+system+with+microcontroller+to+determine+limb+motor+function+of+laboratory+animals%5BTitle%5D)+AND+%22J+Neurosci+Methods%22%5BJournal%5D)">An inclined plane system with microcontroller to determine limb motor function of laboratory animals.</a> J Neurosci Methods. 168 (1), 186-194 (2008).</li>
<li>Gartshore, G., Patterson, J., Macrae, I. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Influence+of+ischemia+and+reperfusion+on+the+course+of+brain+tissue+swelling+and+blood-brain+barrier+permeability+in+a+rodent+model+of+transient+focal+cerebral+ischemia%5BTitle%5D)+AND+%22Exp+Neurol%22%5BJournal%5D)">Influence of ischemia and reperfusion on the course of brain tissue swelling and blood-brain barrier permeability in a rodent model of transient focal cerebral ischemia.</a> Exp Neurol. 147 (2), 353-360 (1997).</li>
<li>Chen, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rodent+stroke+induced+by+photochemical+occlusion+of+proximal+middle+cerebral+artery%3A+Evolution+monitored+with+MR+imaging+and+histopathology%5BTitle%5D)+AND+%22Eur+J+Radiol%22%5BJournal%5D)">Rodent stroke induced by photochemical occlusion of proximal middle cerebral artery: Evolution monitored with MR imaging and histopathology.</a> Eur J Radiol. 63 (1), 68-75 (2007).</li>
<li>Almenning, I., Rieber-Mohn, A., Lundgren, K. M., Lovvik, T. S., Garnaes, K. K., Moholdt, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+high+intensity+interval+training+and+strength+training+on+metabolic%2C+cardiovascular+and+hormonal+outcomes+in+women+with+polycystic+ovary+syndrome%3A+a+pilot+study%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Effects of high intensity interval training and strength training on metabolic, cardiovascular and hormonal outcomes in women with polycystic ovary syndrome: a pilot study.</a> PLoS One. 10 (9), (2015).</li>
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</ol>

The authors have nothing to disclose.

这个协议描述了用于减少在动物中的中风的影响的严重性一种高效运行的轮系统。作为一只老鼠友好的测试平台，该平台是在一个稳定的运行速度，可以由老鼠在整个运行过程中通过预定的自适应加速曲线来保持这样一种方式设计为好。在典型的培训制度，培训预置速度和持续时间手动设置。一旦运动开始，达到预设的速度很快。在这种情况下，这是非常可能的是老鼠都无法达到更高的速度，使?...

笔画持续存在，作为一个财政负担在全球范围内的国家，留下了无数患者的身体和智力残疾1,2。有临床证据表明，经常运动可以改善神经再生和神经加强连接3，4，它也表明，锻炼可以降低患缺血性中风5的风险。无论使用哪种跑步机或正在运行的轮作为运动训练系统，啮齿动物，如老鼠，作为代理人类在绝大多数的临床实验测试6演习的有效性- 8。一个培训系统通常包括训练老鼠在一定时间内，在此期间，大鼠以一定的速度运行。 8 -因此，训练强度是根据运动速度和持续时间6一般计算。同样的方法也适用于估计的运动对神经生理学保护所需的量。然而，实验练习，有时发现是无效的，比如当老鼠绊倒，跌倒，或抓住栏杆，一旦他们无法赶上跑车轮转速9 - 11。不用说，无效运动的事件显著减少运动益处。即使没有任何普遍接受的做法目前量化减少脑损伤的有效练习，有效的锻炼水平依然矗立作为一个客观的评价，为临床研究来说明神经生理学学科锻炼的好处。 存在于在今天的脑损伤减少实验12中使用市售的动物移动系统许多限制。在跑步机上的情况下，老鼠被迫用电击的方式运行，引起了巨大的心理应力的动物，从而干扰在最终神经生理学测试结果8，13，14。运行车轮可分为两种类型，即自愿和被迫的。自愿转轮让老鼠自然运行，创造到期大鼠的身体特征和能力15的差异过大的可变性，而电动行驶轮（MRWs）采用电机转动轮子，迫使老鼠跑。尽管也被强迫训练的一种形式，MRWs强加给大鼠比跑步机13，16，17心理压力较少。然而，使用MRWs实验已经报道，大鼠通过抓住轮轨道上的轨道和拒绝在速度超过20米/分钟9运行有时中断锻炼。这些例子表明，目前可用的动物移动系统具有固有的缺点，即抑制有效锻炼。对于客观鼠训练目的，一种高效的培训体系的发展，但与低干扰，因此被视为神经生理学实验练习一个紧迫的问题。 这项研究提出关于减少的行程11的影响的严重性为实验一种高效运行的轮系统。除了干扰因素的数量减少时的训练过程中，这个系统利用嵌入在轮，从而实现有效的运动活动的更可靠的估计的红外传感器的大鼠的运行位置。由传统的跑步机和在MRWs都频繁行使中断所施加的心理应激歪斜所得运动估计的客观性。在本研究中提出的定位跑步轮（PRW）系统，以试图最小化不必要的干扰发达而用于量化有效的exe提供了可靠的训练模型rcise。

<table><tbody><tr><td>Brushless DC motor</td><td>Oriental Motor</td><td>BLEM512-GFS</td><td></td></tr><tr><td>Motor driver</td><td>Oriental Motor</td><td>BLED12A</td><td></td></tr><tr><td>Motor reducer</td><td>Oriental Motor</td><td>GFS5G20</td><td></td></tr><tr><td>Speedometer</td><td>Oriental Motor</td><td>OPX-2A</td><td></td></tr><tr><td>Treadmill</td><td>Columbus Instruments</td><td>Exer-6M</td><td></td></tr><tr><td>Infrared transmitter&amp;nbsp;</td><td>Seeed Studio</td><td>TSAL6200</td><td></td></tr><tr><td>Infrared Receiver</td><td>Seeed Studio</td><td>TSOP382</td><td></td></tr><tr><td>Microcontroller</td><td>Silicon Labs</td><td>C8051F330</td><td></td></tr><tr><td>CCD camera</td><td>Canon Inc.</td><td>EOS 450D</td><td></td></tr><tr><td>Image processing software</td><td>Adobe Systems Incorporated</td><td>ADOBE Photoshop CS5 12.0</td><td></td></tr><tr><td>Image analysis</td><td>Media Cybernetics</td><td>Pro Plus 4.50.29</td><td></td></tr><tr><td>Sodium pentobarbital</td><td>Sigma-Aldrich (Saint Louis, MO, USA)</td><td>SIGMA P-3761</td><td></td></tr><tr><td>Ketamine</td><td>Pfizer (Kent, UK)</td><td>&amp;nbsp;1867-66-9</td><td></td></tr><tr><td>Atropine</td><td>Taiwan Biotech Co., Ltd. (Taoyuan, Taiwan)</td><td>A03BA01</td><td></td></tr><tr><td>Xylazine</td><td>Sigma-Aldrich (Saint Louis, MO, USA)</td><td>SIGMA X1126</td><td></td></tr><tr><td>Buprenorphine</td><td>Sigma-Aldrich (Saint Louis, MO, USA)</td><td>B9275</td><td></td></tr></tbody></table>

an innovative running wheel-based mechanism for improved rat training performance

这项研究提出的动物移动性系统，配备有定位跑步轮（PRW），作为一种方法来量化的锻炼活动的功效降低的大鼠中风的影响的严重性。该系统提供了比商用系统，如跑步机和电动跑步轮（MRWs）更有效的动物运动训练。与此相反的MRW，只有能够达到以下速度20米/分钟，只被允许在30米的速度稳定运行/分钟由15厘米宽的丙烯酸轮与支持的更宽敞，高密度橡胶跑道一个直径为55厘米的这项工作。使用预定义的自适应加速曲线，该系统不仅降低了操作错误，但也训练的老鼠，直到达到规定的强度持续运行。作为一种方法来评估锻炼效果，大鼠的实时位置被四对部署在跑轮红外线传感器检测到。一旦自适应加速曲线是使用微控制器发起的，由红外传感器得到的数据自动记录和分析的计算机。为了便于比较，3个星期的训练中使用了跑步机，MRW和PRW老鼠进行的。手术诱导大脑中动脉闭塞（MCAO）后，修饰的神经严重性评分（MNSS）和一个倾斜面测试进行了对神经损伤评估给大鼠。 PRW，实验验证作为该动物移动系统中最有效的。此外，运动效果的措施，根据大鼠位置分析，表明存在有效行使和梗死体积之间的高的负相关，并且可用于量化在任何类型的脑减少损伤实验的大鼠的训练。

这部分是专门的比较，手术后制成1周，在MNSS分数，斜面测试结果和脑的梗死体积五组之一。 图4A和图4B分别呈现平均MNSS分数和斜面试验结果的平均值。该PRW组作为最好的出现在MNSS改善方面。 PRW和MRW之间以及跑步机和PRW之间的差异显著清楚地表明，PRW更有效地预防中风比目前市面上其他动物的移动系统。倾斜的平面测试在所有的运动组显著陡倾?...

Watch this Scientific Journal Video about An Innovative Running Wheel-based Mechanism for Improved Rat Training Performance at JoVE.com

An Innovative Running Wheel-based Mechanism for Improved Rat Training Performance

这项研究提出了一个创新型轮运行动物的调动制度的大鼠量化一个有效的锻炼活动。将大鼠友好测试平台构建，使用预定义的自适应加速曲线，并有效地行使率和梗塞体积之间的高相关性表明该协议的中风预防实验潜力。

为提高训练大鼠绩效的创新运行基于轮机制

This study presents an animal mobility system, equipped with a positioning running wheel (PRW), as a way to quantify the efficacy of an exercise activity ...

an-innovative-running-wheel-based-mechanism-for-improved-rat-training

Research

JoVE Journal

Behavior

8.9K 次观看.  National Chin-Yi University of Technology. 这项研究提出了一个创新型轮运行动物的调动制度的大鼠量化一个有效的锻炼活动。将大鼠友好测试平台构建，使用预定义的自适应加速曲线，并有效地行使率和梗塞体积之间的高相关性表明该协议的中风预防实验潜力。 

视频: An Innovative Running Wheel-based Mechanism for Improved Rat Training Performance

A Fully Automated Rodent Conditioning Protocol for Sensorimotor Integration and Cognitive Control Experiments

Rodents have been traditionally used as a standard animal model in laboratory experiments involving a myriad of sensory, cognitive, and motor tasks. Higher cognitive functions that require precise control over sensorimotor responses such as decision-making and attentional modulation, however, are typically assessed in nonhuman primates. Despite the richness of primate behavior that allows multiple variants of these functions to be studied, the rodent model remains an attractive, cost-effective alternative to primate models. Furthermore, the ability to fully automate operant conditioning in rodents adds unique advantages over the labor intensive training of nonhuman primates while studying a broad range of these complex functions.
Here, we introduce a protocol for operantly conditioning rats on performing working memory tasks. During critical epochs of the task, the protocol ensures that the animal's overt movement is minimized by requiring the animal to 'fixate' until a Go cue is delivered, akin to nonhuman primate experimental design. A simple two alternative forced choice task is implemented to demonstrate the performance. We discuss the application of this paradigm to other tasks.

A fully automated protocol for rodent operant conditioning is proposed. The protocol relies on precise temporal control of behavioral events to investigate the extent to which this control influences neural activity underlying sensorimotor integration and cognitive control experiments.

为感觉运动整合与认知控制实验完全自动化的啮齿动物调理协议

Rodents have been traditionally used as a standard animal model in laboratory experiments involving a myriad of sensory, cognitive, and motor tasks. ...

科研

行为学

Kinematics and Ground Reaction Force Determination: A Demonstration Quantifying Locomotor Abilities of Young Adult, Middle-aged, and Geriatric Rats

Behavior, in its broadest definition, can be defined as the motor manifestation of physiologic processes. As such, all behaviors manifest through the motor system. In the fields of neuroscience and orthopedics, locomotion is a commonly evaluated behavior for a variety of disease models. For example, locomotor recovery after traumatic injury to the nervous system is one of the most commonly evaluated behaviors 1-3. Though locomotion can be evaluated using a variety of endpoint measurements (e.g. time taken to complete a locomotor task, etc), semiquantitative kinematic measures (e.g. ordinal rating scales (e.g. Basso Beattie and Bresnahan locomotor (BBB) rating scale, etc)) and surrogate measures of behaviour (e.g. muscle force, nerve conduction velocity, etc), only kinetics (force measurements) and kinematics (measurements of body segments in space) provide a detailed description of the strategy by which an animal is able to locomote 1. Though not new, kinematic and kinetic measurements of locomoting rodents is now more readily accessible due to the availability of commercially available equipment designed for this purpose. Importantly, however, experimenters need to be very familiar with theory of biomechanical analyses and understand the benefits and limitations of these forms of analyses prior to embarking on what will become a relatively labor-intensive study. The present paper aims to describe a method for collecting kinematic and ground reaction force data using commercially available equipment. Details of equipment and apparatus set-up, pre-training of animals, inclusion and exclusion criteria of acceptable runs, and methods for collecting the data are described. We illustrate the utility of this behavioral analysis technique by describing the kinematics and kinetics of strain-matched young adult, middle-aged, and geriatric rats.

Locomotion is often examined as a behavioural outcome in various models of disease in fields such as neuroscience and orthopedics. This video paper intends to describe a method for collecting ground reaction forces and kinematics from rats during unrestrained locomotion.

运动学与地面反作用力的测定：一个示范量化青年，中年和老年大鼠的运动能力能力

Behavior, in its broadest definition, can be defined as the motor manifestation of physiologic processes.  As such, all behaviors manifest through the ...

神经科学

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

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

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

收购高精度熟练的前肢到达任务大鼠

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

Investigating Motor Skill Learning Processes with a Robotic Manipulandum

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

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

调查运动技能的学习过程与机器人Manipulandum

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

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

Tasks that accurately measure dexterity in animal models are critical to understand hand function. Current rat behavioral tasks that measure dexterity largely use video analysis of reaching or food manipulation. While these tasks are easy to implement and are robust across disease models, they are subjective and laborious for the experimenter. Automating traditional tasks or creating new automated tasks can make the tasks more efficient, objective, and quantitative. Since rats are less dexterous than primates, central nervous system (CNS) injury produces more subtle deficits in dexterity, however, supination is highly affected in rodents and crucial to hand function in primates. Therefore, we designed a semi-automated task that measures forelimb supination in rats. Rats are trained to reach and grasp a knob-shaped manipulandum and turn the manipulandum in supination to receive a reward. Rats can acquire the skill within 20 &plusmn; 5 days. While the early part of training is highly supervised, much of the training is done without direct supervision. The task reliably and reproducibly captures subtle deficits after injury and shows functional recovery that accurately reflects clinical recovery curves. Analysis of data is performed by specialized software through a graphical user interface that is designed to be intuitive. We also give solutions to common problems encountered during training, and show that minor corrections to behavior early in training produce reliable acquisition of supination. Thus, the knob supination task provides efficient and quantitative evaluation of a critical movement for dexterity in rats.

This manuscript describes a semi-automated task that quantifies supination in rats. Rats reach, grasp, and supinate a spherical manipulandum. The rat is rewarded with a pellet if the turn angle exceeds a criterion set by the user. This task increases throughput, sensitivity to injury, and objectivity compared to traditional tasks.

旋钮旋任务: 一种半自动评估大鼠前肢功能的方法

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

A Novel Single Animal Motor Function Tracking System Using Simple, Readily Available Software

We have recently demonstrated that implanting intracortical microelectrodes in the motor corteces of rats&#160;results in immediate and lasting motor deficits. Motor impairments were manually quantified through an open field grid test to measure the gross motor function and through a ladder test to measure the fine motor function. Here, we discuss a technique for the automated quantification of the video-recorded tests using our custom Capadona Behavioral Video Analysis System: Grid and Ladder Test, or BVAS. Leveraging simple and readily available coding software (see the Table of Materials), this program allows for the tracking of a single animal on both the open field grid and the ladder tests. In open field grid tracking, the code thresholds the video for intensity, tracks the position of the rat over the 3 min duration of the grid test, and analyzes the path. It then computes and returns measurements for the total distance traveled, the maximum velocity achieved, the number of left- and right-handed turns, and the total number of grid lines crossed by the rat. In ladder tracking, the code again thresholds the video for intensity, tracks the movement of the rat across the ladder, and returns calculated measurements including the time it took the rat to cross the ladder, the number of paw slips occurring below the plane of the ladder rungs, and the incidence of failures due to stagnation or reversals. We envision that the BVAS developed here can be employed for the analysis of motor function in a variety of applications, including many injury or disease models.

The current study aimed to automate the quantification of motor deficits in rats. The initial evaluation model assesses motor loss resulting from an intracortical microelectrode implantation in the motor cortex. We report on the development and use of a tracking algorithm using easily adaptable, simple, and readily available coding software.

一种新的单动物运动功能跟踪系统, 使用简单、易于使用的软件

We have recently demonstrated that implanting intracortical microelectrodes in the motor corteces of rats&#160;results in immediate and lasting motor ...

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

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

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

自动大鼠单丸到达与3维重建的爪子和数字轨迹

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

Utilizing a Reconfigurable Maze System to Enhance the Reproducibility of Spatial Navigation Tests in Rodents

Several maze shapes are used to test spatial navigation performance and behavioral phenotypes. Traditionally, each experiment requires a unique maze shape, thus requiring several separate mazes in different configurations. The maze geometry cannot be reconfigured in a single environment to accommodate scalability and reproducibility. The reconfigurable maze is a unique approach to address the limitations, allowing quick and flexible configurations of maze pathways in a repeatable manner. It consists of interlocking pathways and includes feeders, treadmills, movable walls, and shut-off sensors. The current protocol describes how the reconfigurable maze can replicate existing mazes, including the T-shaped, plus-shaped, W-shaped, and figure-eight mazes. Initially, the T-shaped maze was constructed inside a single experimental room, followed by modifications. The rapid and scalable protocol outlined herein demonstrates the flexibility of the reconfigurable maze, achieved through the addition of components and behavioral training phases in a stepwise manner. The reconfigurable maze systematically and precisely assesses the performance of multiple aspects of spatial navigation behavior.

The present protocol describes a reconfigurable maze, a unique system for testing spatial navigation and behavioral phenotypes in rodents. The adaptability of this maze system enables the execution of various experiments in a single physical environment. The ease of structural rearrangement generates reliable and reproducible experimental results.

利用可重构的迷宫系统提高啮齿动物空间导航测试的可重复性

Several maze shapes are used to test spatial navigation performance and behavioral phenotypes. Traditionally, each experiment requires a unique maze ...

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 ...

使用 Erasmus Ladder 自动评估小鼠的精细运动性能

Monitoring Fine and Associative Motor Learning in Mice Using the Erasmus Ladder

Behavior is shaped by actions, and actions necessitate motor skills such as strength, coordination, and learning. None of the behaviors essential for sustaining life would be possible without the ability to transition from one position to another. Unfortunately, motor skills can be compromised in a wide array of diseases. Therefore, investigating the mechanisms of motor functions at the cellular, molecular, and circuit levels, as well as understanding the symptoms, causes, and progression of motor disorders, is crucial for developing effective treatments. Mouse models are frequently employed for this purpose.
This article describes a protocol that allows the monitoring of various aspects of motor performance and learning in mice using an automated tool called the Erasmus Ladder. The assay involves two phases: an initial phase where mice are trained to navigate a horizontal ladder built of irregular rungs (&#34;fine motor learning&#34;), and a second phase where an obstacle is presented in the path of the moving animal. The perturbation can be unexpected (&#34;challenged motor learning&#34;) or preceded by an auditory tone (&#34;associative motor learning&#34;). The task is easy to conduct and is fully supported by automated software.
This report shows how different readouts from the test, when analyzed with sensitive statistical methods, allow fine monitoring of mouse motor skills using a small cohort of mice. We propose that the method will be highly sensitive to evaluate motor adaptations driven by environmental modifications as well as early-stage subtle motor deficits in mutant mice with compromised motor functions.

作者聚焦：通过使用 Erasmus Ladder 对运动学习和髓鞘可塑性研究进行自动评估来揭示神经机制 

This article presents a protocol that allows a non-invasive and automated assessment of fine motor performance, as well as adaptive and associative motor learning upon challenges, using a device called the Erasmus Ladder. Task difficulty can be titrated to detect motor impairment ranging from major to subtle degrees.

使用伊拉斯谟量程监测小鼠的精细和联想运动学习

Behavior is shaped by actions, and actions necessitate motor skills such as strength, coordination, and learning. None of the behaviors essential for ...