WDA is supported by grant funding from NIH-NICHD (5K12HD001097-17) and Cure SMA. SJK is supported by grant funding from NINDS (K08NS067282 and U01NS079163).

该协议被批准，并粘附在美国俄亥俄州立大学的Wexner医学中心的动物护理和伦理准则。 1.动物准备及麻醉<ol><li>在处理小鼠戴上手套。 </li><li>麻醉小鼠吸入异氟醚和发生在俯卧位。诱导使用3-5％异氟醚和每分钟O 2流量1升麻醉。继诱导麻醉，麻醉维持在2-3％，并每分钟O 2流量1升。 <ol><li>调节O 2流量和异氟烷百分比用于根据动物的疾病状态，年龄，和呼吸率充分麻醉。较小或较弱的动物可能需要适当的麻醉（ 即 1.5-2.5％异氟醚），异氟醚较少。 </li><li>通过施加光后肢脚掌的压力与物体如钳子证明缺乏戒断反应的确认充分麻醉。; </li></ol></li><li>保持温度在37℃的表面温度用的恒温加温板如温度变化​​可能会影响CMAP大小和持续时间。 </li><li>应用兽医石油基软膏的眼睛，以防止干燥。监测麻醉观察呼吸率水平并评估以下压力通过钳子施加到脚垫戒断反应。 </li><li>使用快船待研究后肢清除头发。去除毛发从后肢（S）以进行研究后，轻轻延伸后肢在膝盖处，绑架在臀部和固定到使用胶带的工作表面（ 见图1）。 </li><li>继CMAP和MUNE录音和麻醉停药，不要让动物无人看管，直到它重新获得足够的意识，以保持胸骨斜卧。不返回到动物的其他动物的公司直到完全恢复。 </li></ol> 2。RECORDING设置和设备<ol><li>地方的CMAP和MUNE记录电极如下图如图 1所示。 </li><li>用两个细环状电极的记录电极。 <ol><li>放置活性（E1）环上的皮肤覆盖在后肢的腓肠肌的近侧部分，在膝关节电极，和在皮肤上的脚的中间跖骨部基准（E2）环形电极。 </li><li>为了减少阻抗，外套皮肤环状电极底层用凝胶充分饱和残毛和最大化电极 - 皮肤接触。避免电极凝胶的过量施用，因为这可能会导致电极之间的桥梁，可以防止准确地记录。 </li></ol></li><li>对于坐骨神经的近端后肢刺激，使用两个绝缘28克单极针作为阴极和阳极。插入该阴极在近端后肢的区域并插入肛德更近侧在皮下组织覆盖骶骨。 <ol><li>避免插入刺激电极过于接近坐骨神经或太深，这将直接伤害坐骨神经或其它结构。 图1示出的电极放置。 </li></ol></li><li>为接地电极，放置在对侧后肢或尾部一次性表面电极。 </li></ol> 3.数据采集<ol><li>坐骨神经CMAP <ol><li>通过刺激坐骨神经为0.1毫秒持续时间方波脉冲和强度不等的1-10毫安获得坐骨神经CMAP响应。 </li><li>获得CMAP响应随刺激强度直到不再增加了响应的幅度。然后，为了确保超最大刺激，增加的刺激，以用于获得一个最大应答的刺激强度的〜120％，将获得的附加响应。如果没有进一步增加我n个CMAP大小，记录这个响应，最大CMAP。 </li><li>记录基线至峰和峰-峰振幅CMAP以mV（图2）。 </li></ol></li><li>平均单运动单位电位（SMUP）尺寸和MUNE计算<ol><li>确定平均单个运动单位电位（SMUP）大小的增量增产技术1。为了获得增量响应，提供0.1毫秒的持续时间最大刺激在1赫兹的频率，同时增加了强度0.03毫安步骤获得最小的全或无响应。获得与0.21毫安和0.70 mA的刺激强度的初步反应。 <ol><li>如果初始响应不与0.21 mA和0.70 mA的刺激强度发生，调整刺激阴极位置更近或远离坐骨神经的位置在近端大腿以减少或增加所需的刺激强度分别。 </li><li>如果初始我获得ncremental响应与0.21 mA和0.70 mA的之间的刺激强度，并满足下面指出（3.2.2），商店的标准并记录额外的增量而增加刺激强度是通过以下步骤0.03毫安调整，以获得总共9个附加增量是符合既定标准。 </li></ol></li><li>在增量响应的测量结果，确保每个增量满足以下条件。 <ol><li>确保增量响应的初始负峰值在时间上显示为CMAP说明的图 2中的阴影部分的最大CMAP响应的负峰值内对齐。 </li><li>确保每个增量响应是稳定的，没有分馏，通过观察三个重复反应确立。区分实时（叠加在先前记录单位）目视增量响应。 注:每个增量应该是视觉上不同并与前述的反应（图3）放大比较。分析实时允许识别较大振幅（增量值）的反应相比于现有的反应，和归属于背景噪声小的变化可以忽略不计。增量为10甲叠加图示于图4（B和D），以进一步说明这一点。 </li><li>后目视确认每个增量，确保所测量的振幅差（确认响应振幅的现有响应=振幅差）为至少25μV。 </li><li>如果增量小于25μV，丢弃和重新测量的响应。记录10增量后的反应，评估增量，以确保每个单独的增量响应的幅度不大于所有十个的增量的总和 （即最终响应的总幅度）的1/3。如果这个条件不满足，重新测量增量响应。 </li></ol></li><li>平均10个增量值，得到平均单个马达单元电位（SMUP）振幅的估计（图3）。注意: 图3详细说明了平均SMUP计算的基础，但平均SMUP振幅可以由最终增量响应的整个振幅除以增量的总数来简单地计算（即，10）。 实施例个别SMUP计算（图3中示出）: SMUP 1 =峰 - 峰值增量1的振幅 SMUP 1 = 0.050毫伏 SMUP 2 =（峰 - 峰振幅增量2） - （增量峰 - 峰振幅1） SMUP 2 = 0.150毫伏0.050 MV = 0.100毫伏<ol><li>计算每个后续的递增（最多总共10），并进行平均十个的增量。 </li></ol></li><li>计算MUNE由平均SMUP振幅除以最大CMAP波幅（峰 - 峰）（峰到峰）。 （MUNE = CMAP /平均SMUP）。在一些电系统中，SMUP增量测量μV，而CMAP在毫伏一般规定。必要时，转换CMAP和SMUP结果之前MUNE计算类似的单位。 </li></ol></li></ol>

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The authors have nothing to disclose.

MUNE和CMAP经常用在调查研究和监测患者的神经肌肉疾病如ALS和脊髓性肌萎缩（SMA）9,10临床相关的措施，例如，在形状记忆合金，CMAP和MUNE很好地相关与年龄，严重性和临床功能10-14措施。两个措施是微创和允许的功能评估纵向在同一个人。重要的是，这些措施不能测量活化或招募电动机单元的皮质运动神经元，但它们提供的运动神经元的完整性和其功能对应，电动机单元一个临床相关的评估。 神经肌肉疾病的动物模型是人类疾病的发病机制的理解以及对潜在的有效治疗剂的临床前的发展是至关重要的。翻译结果的措施和生物标志物，可以的能力跨物种的利用可以促进和加快有前途的临床前研究结果对人类临床试验的翻译。几个研究小组先前已经利用两个电和力（机械）的测量来估算运动单位的功能在小鼠模型2-4，15-22。由于措施的相对复杂性，我们还改进了直观的格式，这些技术允许更广泛地使用和实施的小鼠。视频演示和指令的格式，允许强调过程的关键步骤和潜在的缺陷加以解决。这些技术在运动神经元疾病的潜在疗法的临床前试验中的应用可能会提高翻译公认的治疗方法，从小鼠到人类疾病。 有收购的CMAP和MUNE反应的过程中的几个关键步骤。正确的和一致的记录电极放置和用后腿足够的电极接触肢是幅度和减小背景噪声重现的测量是至关重要的。因此，后肢皮肤和电极之间的密切接触，应始终如一地证实。我们已发现，表面电极提供更加一致和CMAP MUNE录音比针电极。由于非常薄的皮下组织，针记录表面的小的移动可导致在CMAP的幅度很大的差异。此外，针状电极更侵入性是不是最佳的新生小鼠或纵向研究，由于潜在的肌肉破坏和伤害。的非选择性，表面电极记录一个潜在缺点涉及的减弱表现型分辨率的可能性，如果一个特定肌肉是或多或少参与与另一个相比，这已被报道的ALS小鼠模型21。 相比于CMAP获取平均SMUP尺寸在技术上是更具挑战性。由于较小responsE尺寸（以μV而非毫伏的范围）的背景噪声可以更成问题。背景噪声可以通过调整接地电极，阴极，阳极，和检查等电气设备的实验装置附近被减小。法拉第笼，通常用于细胞内电应用中，并不需要。视觉确定个人SMUP反应是最困难的技巧掌握，并采取实践有足够的可重复性一致的结果。以确保正在记录SMUPs最大CMAP响应的持续时间内启动是重要的。我们已经确定的标准验收的各个增量反应，使这个过程更易于执行，增加区域内和跨信度。 增量MUNE技术的一个潜在的缺点包括高估的功能的运动单位的数量，由于摩托车的交替的可能性R单元。我们已经使用类似于Shefner 等人的技术。在每个反应，应再现地看出，共3次，以减少这种现象3的影响。 根据我们的经验，临床电生理系统本文所述由于改善考官，电学系统接口的人体工程学设计，允许容易控制的研究进行了优化。在我们的实验室中使用的双通道系统配备有使用的放大器具有24比特模拟到数字转换器和每个通道48 kHz的采样率的两个非切换放大器通道。硬件增益可以从10nV调整到100 mV /分。低频滤波器具有范围从0.2赫兹-5千赫和高频滤波器设置范围从30赫兹-10千赫。恒定电流刺激时（强度:0〜100毫安，持续时间:0.02-1毫秒）。大多数临床系统具有类似的适当的功能，并且可以进行调整，以充分地记录和CMAP MUNE响应。一个dditionally，标准电生理钻机可组装到适当记录和CMAP MUNE，但接口可能需要调整为便于刺激调整和快速识别和CMAP SMUP响应。 我们以前曾利用CMAP的技术和MUNE这里描述以允许在产后早期到成年5在小鼠后肢的坐骨神经神经支配的肌肉的快速和可重复的评估。这些技术允许评估在小鼠模型中当行为测试为运动功能是不可行的或者是不可靠的。促进这种技术新生小鼠应用运动单位发展的研究，并有扩大我们的运动神经元的神经支配和修剪理解的潜力。例如，我们已经表明，记录有MUNE官能马达单元的数量将在头两个个星期从polyneuronal修剪到mononeuronal神经支配期间增加生活S在新生小鼠5。测试小鼠过长时间使用这种技术的能力适合于电机单元响应于外周神经损伤，遗传性神经肌肉疾病和老化的研究。

马达单元数估计（MUNE）最初是由麦科马斯等人所述 。超过三十年以前1。原来的技术是，使用的刺激逐渐增加，以获得次最大增量的复合肌肉动作电位（CMAP）记录技术的改进。这些增量进行相加并进行平均，以确定一个马达单元电位（SMUP）的估计大小。这个大小被分为CMAP响应来估计电动机单元的数目支配被测试的肌肉。以下的原始描述，同时使用电生理反应和增量力许多变化（机械）测量已用于人类研究和动物模型2。该MUNE技术是由Shefner和同事改性调查肌萎缩性侧索硬化症（ALS）3，4的小鼠模型。 在当前的描述中，我们详细介绍simplif的MUNE技术是快速执行IED修改。重要的是，CMAP和MUNE允许在这两个新生和成年小鼠5-8可靠的措施。有经验的人可以在每头牲畜10-20分钟执行这些措施，并重复措施是允许收购纵向数据5可行的。在目前的研究中，我们采用了临床电诊断系统。根据我们的经验，临床电生理系统在体内电生理数据的快速，高效地捕捉优化，但标准的电钻机可以很容易地适应这种应用。

<table><tbody><tr><td>Pro trimmer Pet Grooming Kit</td><td>Oster</td><td>078577-010-003</td><td>clippers for hair removal</td></tr><tr><td>Synergy T2 EMG system</td><td>Natus Neurology</td><td>Model no longer available</td><td>portable electrodiagnostic system</td></tr><tr><td>monopolar needles 28 G</td><td>Teca</td><td>017K121</td><td>cathode and anode stimulating electrodes</td></tr><tr><td>Alpine Biomed Digital Ring Electrode with twisted wires and 1.5 mm TP connectors.</td><td>Alpine Biomed</td><td>9013S0312</td><td>recording electrodes</td></tr><tr><td>Helping Hands alligator clip with iron base</td><td>Radio Shack</td><td>64-079</td><td>Maintaining recording electrode placement&amp;nbsp;</td></tr><tr><td>Spectra 360 Electrode Gel&amp;nbsp;</td><td>Parker Laboratories</td><td>9013G5012</td><td>applied to reduce skin impedance</td></tr><tr><td>monoject curved tip irrigating syringe</td><td>Covidien</td><td>81412012</td><td>utilized for application of electrode gel</td></tr><tr><td>EMG needle cable</td><td>Teca</td><td>902-RLC-TP&amp;nbsp;</td><td>to connect monopolar electrodes to electrodiagnostic stimulator</td></tr><tr><td>Disposable 2&quot; x 2&quot; Electrode or similar trimmed as needed</td><td>Carefusion</td><td>019-415000&amp;nbsp;</td><td>ground electrode</td></tr><tr><td>Small Heating Plate with built-in RTD sensor, 15 x10 cm</td><td>World Precision Instruments</td><td>61830</td><td>warming plate used with animal temperature controller to transmit heat to animal</td></tr><tr><td>Silicone pad for use with ATC2000</td><td>World Precision Instruments</td><td>503573</td><td>conductive removable pad to cover warming plate for easy cleaning</td></tr><tr><td>Animal temperature controller</td><td>World Precision Instruments</td><td>ATC2000</td><td>low noise animal heating system for maintaining animal temperature</td></tr><tr><td>Veterinarian petroleum-based ophthalmic ointment&amp;nbsp;</td><td>Puralube</td><td>26870</td><td>applied during anesthesia to avoid corneal injury</td></tr></tbody></table>

electrophysiological motor unit number estimation (mune) measuring compound muscle action potential (cmap) in mouse hindlimb muscles

Compound muscle action potential (CMAP) and motor unit number estimation (MUNE) are electrophysiological techniques that can be used to monitor the functional status of a motor unit pool in vivo. These measures can provide insight into the normal development and degeneration of the neuromuscular system. These measures have clear translational potential because they are routinely applied in diagnostic and clinical human studies. We present electrophysiological techniques similar to those employed in humans to allow recordings of mouse sciatic nerve function. The CMAP response represents the electrophysiological output from a muscle or group of muscles following supramaximal stimulation of a peripheral nerve. MUNE is an electrophysiological technique that is based on modifications of the CMAP response. MUNE is a calculated value that represents the estimated number of motor neurons or axons (motor control input) supplying the muscle or group of muscles being tested. We present methods for recording CMAP responses from the proximal leg muscles using surface recording electrodes following the stimulation of the sciatic nerve in mice. An incremental MUNE technique is described using submaximal stimuli to determine the average single motor unit potential (SMUP) size. MUNE is calculated by dividing the CMAP amplitude (peak-to-peak) by the SMUP amplitude (peak-to-peak). These electrophysiological techniques allow repeated measures in both neonatal and adult mice in such a manner that facilitates rapid analysis and data collection while reducing the number of animals required for experimental testing. Furthermore, these measures are similar to those recorded in human studies allowing more direct comparisons.

CMAP和MUNE的技术本报告中所述的记录允许利用微创电极放置（图1）的坐骨神经支配后肢肌肉的神经肌肉功能的。超最大CMAP大小，其表示从一肌群的输出总，可以使用振幅和区（图2）的参数进行描述，但是，在目前的方法中，我们使用振幅量化CMAP和SMUP尺寸。由于CMAP应对措施累加肌纤维的去极化肌肉内，病理从运动神经元的肌纤维随时随地都可以导致减少CMAP大小。因此，CMAP给人总的功能状态极好的措施。正如预期的那样，CMAP规模将发展5期间增加。由于可能发生以下去神经（即侧支出芽）代偿性改变，CMAP大小可被保持，尽管马达neur的流程或运动性轴索损失。因此，MUNE的技术中是必需的，以确定运动神经元轴突或输入到肌肉或肌肉群的被测试。个别增量记录（ 图3）允许估计单电机单元（SMUP尺寸）的平均输出的，得到约运动单位的功能状态的更详细信息。 CMAP和MUNE可以用来测量在神经肌肉疾病的各种小鼠模型神经肌肉功能。 在图4中，在成年对照小鼠和成年。鼠标11周后坐骨神经的调查结果进行了对比。继坐骨神经，宗严重降低50估计功能的运动单位与278功能的运动单位在对照组小鼠正常的结果进行比较。与此相反，CMAP波幅在压碎的动物（39.6毫伏基线至峰，74.9毫伏峰 - 峰值）是表示仅轻微减少与对照相比（49.0毫伏基线到- 峰值，84.2毫伏峰 - 峰）由于侧支出芽。 <img alt="图1" src="/files/ftp_upload/52899/52899fig1.jpg" /> 图1.电极放置的黑（E1）"激活"电极（A）和红色（E2）"参考"记录电极（B）的放置在腓肠肌在腓肠肌在膝盖处的近端部分。上述刺激阴极（黑色）（C）和阳极（红色）（D）中被插入皮下近端到记录电极，以产生远端响应。一次性盘电极（D）放置在后肢，尾骶骨或作为地面，以减少工件。 <a href="https://www.jove.com/files/ftp_upload/52899/52899fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 图2.复合肌肉动作电位，图为是代表性的CMAP响应的示图:（A）基准-峰振幅是从等电基线测量到初始负峰值（负电压基线以上描绘）。（ B）中的峰-峰振幅是从负峰 ​​值电压到正峰值电压测量。灰色阴影区为负峰面积。 <a href="https://www.jove.com/files/ftp_upload/52899/52899fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/52899/52899fig3.jpg" /> 图3.增量反应。两个有代表性的增量响应如图叠加和伊索拉化。对于每个增量的MUNE计算幅度被测定的峰 - 峰值。增量＃1是初始全或无响应记录和表示单个马达单元电位（SMUP）。每个后续增量（＃2-10）代表增加量子叠加在之前的回应。因此，为了获得SMUP幅度的递增2-10，现有响应的幅度从获得的增值幅度减去。 <a href="https://www.jove.com/files/ftp_upload/52899/52899fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/52899/52899fig4.jpg" /> 图4.实施例坐骨神经CMAP和MUNE。（A）的坐骨神经复合肌肉动作电位（CMAP）的成人（6月龄）对照小鼠与基线至峰振幅的49.0毫伏和峰到84.2 mV的峰值。屏幕灵敏度=每格10毫伏和屏幕持续时间10毫秒。（B）的十个相应的增量的反应（对照小鼠）与3.028毫伏总幅度被除以10，以确定平均SMUP尺寸（0.3028毫伏）。屏幕的灵敏度= 0.5毫伏和每格1毫秒的扫描速度。计算MUNE = 278（MUNE = CMAP /平均SMUP（84.2毫伏/ 0.3028毫伏））（C）坐骨神经CMAP11周以下的成年小鼠坐骨神经（6个月以下），显示轻度降低基线到峰值（ 39.6毫伏）和峰 - 峰幅度（74.9毫伏）。屏幕灵敏度=每分和每格1毫秒扫描速度为10mV。（D）的十个相应的增量响应（在小鼠与神经压迫）与14.923毫伏总峰-峰振幅除以10获得平均SMUP的1.4923 mV的大小。屏幕的灵敏度= 2每分毫伏和每格1 ms的扫描速度。计算MUNE = 50（MUNE = CMAP/平均SMUP（74.9毫伏/ 1.4923毫伏））。 （**注不同的灵敏度坐骨神经挤压鼠标的增量答复）。 <a href="https://www.jove.com/files/ftp_upload/52899/52899fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Electrophysiological Motor Unit Number Estimation MUNE Measuring Compound Muscle Action Potential CMAP in Mouse Hindlimb Muscles at JoVE.com

Electrophysiological Motor Unit Number Estimation MUNE Measuring Compound Muscle Action Potential CMAP in Mouse Hindlimb Muscles

我们提出精致的协议，允许在电动机单元功能的小鼠体内监测。技术测量复合肌肉动作电位（CMAP）和电动机单元数估计（MUNE）在坐骨神经支配鼠标后肢肌肉进行说明。

电电机组数估计（MUNE）在小鼠后肢肌肉测量复合肌肉动作电位（CMAP）

Compound muscle action potential (CMAP) and motor unit number estimation (MUNE) are electrophysiological techniques that can be used to monitor the ...

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Electrophysiological Motor Unit Number Estimation (MUNE) Measuring Compound Muscle Action Potential (CMAP) in Mouse Hindlimb Muscles

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21.0K 次观看.  The Ohio State University Wexner Medical Center. 我们提出精致的协议，允许在电动机单元功能的小鼠体内监测。技术测量复合肌肉动作电位（CMAP）和电动机单元数估计（MUNE）在坐骨神经支配鼠标后肢肌肉进行说明。 

视频: Electrophysiological Motor Unit Number Estimation MUNE Measuring Compound Muscle Action Potential CMAP in Mouse Hindlimb Muscles

Simultaneous Intracellular Recording of a Lumbar Motoneuron and the Force Produced by its Motor Unit in the Adult Mouse In vivo

The spinal motoneuron has long been a good model system for studying neural function because it is a neuron of the central nervous system with the unique properties of (1) having readily identifiable targets (the muscle fibers) and therefore having a very well-known function (to control muscle contraction); (2) being the convergent target of many spinal and descending networks, hence the name of "final common pathway"; and (3) having a large soma which makes it possible to penetrate them with sharp intracellular electrodes. Furthermore, when studied in vivo, it is possible to record simultaneously the electrical activity of the motoneurons and the force developed by their muscle targets. Performing intracellular recordings of motoneurons in vivo therefore put the experimentalist in the unique position of being able to study, at the same time, all the compartments of the "motor unit" (the name given to the motoneuron, its axon, and the muscle fibers it innervates1): the inputs impinging on the motoneuron, the electrophysiological properties of the motoneuron, and the impact of these properties on the physiological function of the motoneurons, i.e. the force produced by its motor unit. However, this approach is very challenging because the preparation cannot be paralyzed and thus the mechanical stability for the intracellular recording is reduced. Thus, this kind of experiments has only been achieved in cats and in rats. However, the study of spinal motor systems could make a formidable leap if it was possible to perform similar experiments in normal and genetically modified mice.
For technical reasons, the study of the spinal networks in mice has mostly been limited to neonatal in vitro preparations, where the motoneurons and the spinal networks are immature, the motoneurons are separated from their targets, and when studied in slices, the motoneurons are separated from most of their inputs. Until recently, only a few groups had managed to perform intracellular recordings of motoneurons in vivo2-4 , including our team who published a new preparation which allowed us to obtain very stable recordings of motoneurons in vivo in adult mice5,6. However, these recordings were obtained in paralyzed animals, i.e. without the possibility to record the force output of these motoneurons. Here we present an extension of this original preparation in which we were able to obtain simultaneous recordings of the electrophysiological properties of the motoneurons and of the force developed by their motor unit. This is an important achievement, as it allows us to identify the different types of motoneurons based on their force profile, and thereby revealing their function. Coupled with genetic models disturbing spinal segmental circuitry7-9, or reproducting human disease10,11, we expect this technique to be an essential tool for the study of spinal motor system.

Simultaneous Intracellular Recording of a Lumbar Motoneuron and the Force Produced by its Motor Unit in the Adult Mouse In vivo

This new method permits the simultaneous intracellular recording of a single adult mouse motoneuron and the measurement of the force produced by its muscle fibers. The combined investigation of the electrical and mechanical properties of motor units in normal and genetically modified animals is a breakthrough for the study of the neuromuscular system.

同时细胞内记录的车组在成年小鼠的腰椎运动神经元产生的力<em在体内</em

The  spinal motoneuron has long been a good model system for studying neural  function because it is a neuron of the central nervous system with the ...

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神经科学

Assessment of Neuromuscular Function Using Percutaneous Electrical Nerve Stimulation

Percutaneous electrical nerve stimulation is a non-invasive method commonly used to evaluate neuromuscular function from brain to muscle (supra-spinal, spinal and peripheral levels). The present protocol describes how this method can be used to stimulate the posterior tibial nerve that activates plantar flexor muscles. Percutaneous electrical nerve stimulation consists of inducing an electrical stimulus to a motor nerve to evoke a muscular response. Direct (M-wave) and/or indirect (H-reflex) electrophysiological responses can be recorded at rest using surface electromyography. Mechanical (twitch torque) responses can be quantified with a force/torque ergometer. M-wave and twitch torque reflect neuromuscular transmission and excitation-contraction coupling, whereas H-reflex provides an index of spinal excitability. EMG activity and mechanical (superimposed twitch) responses can also be recorded during maximal voluntary contractions to evaluate voluntary activation level. Percutaneous nerve stimulation provides an assessment of neuromuscular function in humans, and is highly beneficial especially for studies evaluating neuromuscular plasticity following acute (fatigue) or chronic (training/detraining) exercise.

We present a protocol to assess changes in neuromuscular function. Percutaneous electrical nerve stimulation is a non-invasive method that evokes muscular responses. Electrophysiological and mechanical properties of these responses permit the evaluation of neuromuscular function from brain to muscle (supra-spinal, spinal and peripheral levels).

神经肌肉功能的应用经皮神经电刺激评估

Percutaneous electrical nerve stimulation is a non-invasive method commonly used to evaluate neuromuscular function from brain to muscle (supra-spinal, ...

CMAP Scan MUNE (MScan) - A Novel Motor Unit Number Estimation (MUNE) Method

Like other methods for motor unit number estimation (MUNE), compound muscle action potential (CMAP) scan MUNE (MScan) is a non-invasive electrophysiologic method to estimate the number of functioning motor units in a muscle. MUNE is an important tool for the assessment of neuropathies and neuronopathies. Unlike most MUNE methods in use, MScan assesses all the motor units in a muscle, by fitting a model to a detailed stimulus-response curve, or CMAP scan. It thereby avoids the bias inherent in all MUNE methods based on extrapolating from a small sample of units. Like &#39;Bayesian MUNE,&#39;&#160;MScan analysis works by fitting a model, made up of motor units with different amplitudes, thresholds, and threshold variabilities, but the fitting method is quite different, and completed within five minutes, rather than several hours. The MScan off-line analysis works in two stages: first, a preliminary model is generated based on the slope and variance of the points in the scan, and second, this model is then refined by adjusting all the parameters to improve the fit between the original scan and scans generated by the model.
This new method has been tested for reproducibility and recording time on 22 amyotrophic lateral sclerosis (ALS) patients and 20 healthy controls, with each test repeated twice by two blinded physicians. MScan showed excellent intra- and inter-rater reproducibility with ICC values of &#62;0.98 and a coefficient of variation averaging 12.3 &#177; 1.6%. There was no difference in the intra-rater reproducibility between the two observers. Average recording time was 6.27 &#177; 0.27 min.
This protocol describes how to record a CMAP scan and how to use the MScan software to derive an estimate of the number and sizes of the functioning motor units. MScan is a fast, convenient, and reproducible method, which may be helpful in diagnoses and monitoring disease progression in neuromuscular disorders.

CMAP Scan MUNE MScan - A Novel Motor Unit Number Estimation MUNE Method

This protocol describes a new method to estimate the number of functioning motor units in a muscle, by fitting a model to a detailed stimulus-response curve of the compound muscle action potential. It is quick and easy to perform and analyze and has excellent reproducibility.

CMAP 扫描免疫性 (MScan)-一种新的电机单元数估计 (免疫性) 方法

Like other methods for motor unit number estimation (MUNE), compound muscle action potential (CMAP) scan MUNE (MScan) is a non-invasive electrophysiologic ...

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

Assessing the functionality of the nerve axon provides detailed information on the progression of neuromuscular disorders. Electrophysiological recordings provide a sensitive approach to measure nerve conduction in humans and rodent models. To broaden the technical possibilities for electromyography in mice, the measurement of compound muscle action potentials (CMAPs) from the brachial plexus nerve in the forelimb using needle electrodes is described here. CMAP recordings after stimulating the sciatic nerve in hindlimbs have been previously described. The newly introduced method here allows for the evaluation of the nerve conductivity at an additional site, and thus provides a more profound overview of the neuromuscular functionality. The technique provides information on both the relative number of functional axons and the myelination level. Thereby, this method can be applied to assess both axonal diseases as well as demyelinating conditions. This minimally invasive method does not require extraction of the nerve and therefore it is suitable for repeated measurements for longitudinal follow-up in the same animal. Similar recordings are performed in clinical setups to emphasize the translational relevance of the method.

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

The measurement of nerve conduction is a useful tool to assess mouse models of neurodegeneration but it is frequently only applied to stimulate the sciatic nerve in hindlimbs. Here, we describe a technique to measure compound muscle action potential (CMAP) in vivo in the mouse forelimb muscles innervated by the brachial plexus.

体内运动神经元变性小鼠前肢肌作用电位的电生理测量

Assessing the functionality of the nerve axon provides detailed information on the progression of neuromuscular disorders. Electrophysiological recordings ...

Non-invasive Assessment of Dorsiflexor Muscle Function in Mice

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively affect skeletal muscle. This can result in a loss of muscle mass (atrophy) and/or loss of muscle quality (reduced force per unit of muscle mass), both of which are prevalent in chronic disease, muscle-specific disease, immobilization, and aging (sarcopenia). Skeletal muscle function in animals can be evaluated by a range of different tests. All tests have limitations related to the physiological testing environment, and the selection of a specific test often depends on the nature of the experiments. Here, we describe an in vivo, non-invasive technique involving a helpful and easy assessment of force frequency-curve (FFC) in mice that can be performed on the same animal over time. This permits monitoring of disease progression and/or efficacy&#160;of a potential therapeutic treatment.

Measurement of rodent skeletal muscle contractile function is a useful tool that can be used to track disease progression as well as efficacy of therapeutic intervention. We describe here the non-invasive, in vivo assessment of the dorsiflexor muscles that can be repeated over time in the same mouse.

小鼠肌功能的无创评价

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively ...

医学

Functional and Physiological Methods of Evaluating Median Nerve Regeneration in the Rat

The main goal of this investigation is to show how to create and repair different types of median nerve (MN) lesions in the rat. Moreover, different methods of simulating postoperative physiotherapy are presented. Multiple standardized strategies are used to assess motor and sensory recovery using an MN model of peripheral nerve lesion and repair, thus permitting easy comparison of the results. Several options are included for providing a postoperative physiotherapy-like environment to rats that have undergone MN injuries. Finally, the paper provides a method to evaluate the recovery of the MN using several noninvasive tests (i.e., grasping test, pin prick test, ladder rung walking test, rope climbing test, and walking track analysis), and physiological measurements (infrared thermography, electroneuromyography, flexion strength evaluation, and flexor carpi radialis muscle weight determination). Hence, this model seems particularly appropriate to replicate a clinical scenario, facilitating extrapolation of results to the human species.
Although the sciatic nerve is the most studied nerve in peripheral nerve research, analysis of the rat MN presents various advantages. For example, there is a reduced incidence of joint contractures and automutilation of the affected limb in MN lesion studies. Furthermore, the MN is not covered by muscle masses, making its dissection easier than that of the sciatic nerve. In addition, MN recovery is observed sooner, because the MN is shorter than the sciatic nerve. Also, the MN has a parallel path to the ulnar nerve in the arm. Hence, the ulnar nerve can be easily used as the nerve graft for repairing MN injuries. Finally, the MN in rats is located in the forelimb, akin to the human upper limb; in humans, the upper limb is the site of most peripheral nerve lesions.

Presented is a protocol to produce different types of median nerve (MN) lesions and repair in the rat. Additionally, the protocol shows how to evaluate the functional recovery of the nerve using several noninvasive behavioral tests and physiological measurements.

大鼠中位神经再生评价的功能与生理方法

The main goal of this investigation is to show how to create and repair different types of median nerve (MN) lesions in the rat. Moreover, different ...

Functional Isolation of Single Motor Units of Rat Medial Gastrocnemius Muscle

This work outlines functional isolation of motor units (MUs), a standard electrophysiological method for determining characteristics of motor units in hindlimb muscles (such as the medial gastrocnemius, soleus, or plantaris muscle) in experimental rats. A crucial element of the method is the application of electrical stimuli delivered to a motor axon isolated from the ventral root. The stimuli may be delivered at constant or variable inter-pulse intervals. This method is suitable for experiments on animals at varying stages of maturity (young, adult or old). Moreover, this protocol can be used in experiments studying variability and plasticity of motor units evoked by a large spectrum of interventions. The results of these experiments may both augment basic knowledge in muscle physiology and be translated into practical applications. This procedure focuses on the surgical preparation for the recording and stimulation of MUs, with an emphasis on the necessary steps to achieve preparation stability and reproducibility of results.

This method allows the recording of the force of twitch and tetanic contractions and action potentials in three types of motor units in the rat medial gastrocnemius muscle. The functional isolation of a single motor unit is induced by electrical stimulation of the axon.

大鼠中胃肌肉单运动单元的功能隔离

This work outlines functional isolation of motor units (MUs), a standard electrophysiological method for determining characteristics of motor units in ...

<meta charset="utf8"></head><body>量化神经退行性啮齿动物模型中的膈运动单位功能

Assessing Rat Diaphragm Motor Unit Connectivity Outcome Measures as Quantitative Biomarkers of Phrenic Motor Neuron Degeneration and Compensation

Loss of ventilatory muscle function is a consequence of motor neuron injury and neurodegeneration (e.g., cervical spinal cord injury and amyotrophic lateral sclerosis, respectively). Phrenic motor neurons are the final link between the central nervous system and muscle, and their respective motor units (groups of muscle fibers innervated by a single motor neuron) represent the smallest functional unit of the neuromuscular ventilatory system. Compound muscle action potential (CMAP), single motor unit potential (SMUP), and motor unit number estimation (MUNE) are established electrophysiological approaches that enable the longitudinal assessment of motor unit integrity in animal models over time but have mostly been applied to limb muscles. Therefore, the objectives of this study are to describe an approach in preclinical rodent studies that can be used longitudinally to quantify the phrenic MUNE, motor unit size (represented as SMUP), and CMAP, and then to demonstrate the utility of these approaches in a motor neuron loss model. Sensitive, objective, and translationally relevant biomarkers for neuronal injury, degeneration, and regeneration in motor neuron injury and diseases can significantly aid and accelerate experimental research discoveries to clinical testing.

<meta charset="utf8"></head><body>作者聚焦：研究神经退行性疾病中的神经肌肉反应和运动神经元可塑性

In this study, we present an in vivo method for estimating motor unit number and size to quantify rat diaphragm motor unit connectivity. A step-by-step approach to these techniques is described.

评估大鼠膈肌运动单元连接结果测量作为膈运动神经元变性和代偿的定量生物标志物

Loss of ventilatory muscle function is a consequence of motor neuron injury and neurodegeneration (e.g., cervical spinal cord injury and amyotrophic ...

电电机组数估计（MUNE）在小鼠后肢肌肉测量复合肌肉动作电位（CMAP）

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同时细胞内记录的车组在成年小鼠的腰椎运动神经元产生的力

神经肌肉功能的应用经皮神经电刺激评估

CMAP 扫描免疫性 (MScan)-一种新的电机单元数估计 (免疫性) 方法

体内运动神经元变性小鼠前肢肌作用电位的电生理测量

小鼠肌功能的无创评价

大鼠中位神经再生评价的功能与生理方法

大鼠中胃肌肉单运动单元的功能隔离

评估大鼠膈肌运动单元连接结果测量作为膈运动神经元变性和代偿的定量生物标志物

评估大鼠膈肌运动单元连接结果测量作为膈运动神经元变性和代偿的定量生物标志物

评估大鼠膈肌运动单元连接结果测量作为膈运动神经元变性和代偿的定量生物标志物