Name: measuring progressive neurological disability in a mouse model of multiple sclerosis
Uploaded: 2016-11-14T13:00:00.000+00:00
Duration: 8 min 11 s
Description: Watch this Scientific Journal Video about Measuring Progressive Neurological Disability in a Mouse Model of Multiple Sclerosis at JoVE.com

The authors thank the staff of the Center for Comparative Medicine and Research (CCMR) at Dartmouth for their expert care of the mice used for these studies. The authors also acknowledge Emily Clough for her excellent administrative support.

所有的动物利用工作的机构动物护理和使用委员会（IACUC）在医学盖泽尔学院达特茅斯审查和批准协议。 1.小鼠模型<ol><li> TMEV诱导脱髓鞘疾病的诱导 <ol><li>移动包含从机架4-6周龄的雌性SJL / JHan小鼠舒适的工作空间中的笼中。标记小鼠（ 例如，用耳标或耳冲），以允许临床和组织学疾病的个体的评价。 </li><li>绘制30微升TMEV感染库存（2×10 6噬斑形成单位; PFU）的在PBS中成29隔距胰岛素注射器和针头。 </li><li>准备麻醉气体机:检查系统，以保证足量的氧气和异氟烷的过程期间存在。 </li><li>打开流量计1升/分钟。放置动物进入吸气室和密封的顶部。打开VAporizer至3.5％和监视动物直到横卧。 </li><li>从室取出的动物，捏住脚垫，保证足够的麻醉测试鼠标。缺乏应对强大的捏表示充分麻醉。 </li><li>清洁注射部位用70％的异丙醇。 </li><li>注入30微升TMEV感染股票的成徒手注射（ 图1）的右大脑半球。注射的部位是眼睛和耳朵线之间，只是离中线大约一半。 </li><li>返回鼠标其控股笼一次高度警惕和移动（通常为3 - 5分钟）。 </li><li>安乐死通过放血或心脏灌注小鼠TMEV感染后3至6个月，这取决于疾病的发展的速度。 </li></ol></li></ol> 2.转棒分析<ol><li> 该转棒仪 <ol><li>之前TMEV感染小鼠的试验与以他们熟悉机，并评估其正常基线平衡协调和电机控制。 </li><li>开始在5日的适配协议感染后（DPI， 即 TMEV感染前5天）。 </li><li>允许小鼠适应到测试室转棒至少30分钟前的测试，以便使它们能够适应环境。 </li><li>确保两个旋转杆单元和计算机已经插入并彼此连接（ 图2）。 </li><li>预先设定的旋转杆与-5 dpi的训练协议参数，如表1所述。 </li><li>保存日期和标识信息的工作文件。 </li><li>移动包含小队要从机架测试邻近于旋转杆的表笼。小鼠通常在4小队进行测试。 </li><li>由尾拿起小鼠，并将其放置在杆，面向远离操作者。重复到第四鼠标的第二位。如果鼠标FALLS或跳转，将它放回到它的旋转杆车道，直到所有的小鼠在适当的位置。如果忽略任何老鼠转身面对运营商。 </li><li>加载所有的老鼠后，按"Enter"键开始实验。观察计时器自动启动，每展出每条车道分钟（RPM）的旋转。 <ol><li>由于每个动物从杆下降时，记录在秋天时的杆的速度，以及时间的持续时间的动物保持在杆上。该杆将继续转动，直到最后动物已经从杆组件下降。 </li></ol></li><li>之后所有的老鼠都下降了，用纸巾从杆删除任何粪便勃利和尿液。尿液和粪便材料的存在可能会影响小鼠握棒的能力。 <ol><li> 3分钟休息后，给小鼠第二次，然后第三个审判。每个单次试验的最大时间为240秒。在每个测试日辖共3项试验。 </li></ol></li><li>返回老鼠的笼子，并返回他们回到机架中。在实验期结束时，清洗旋转杆用肥皂和水去除所有排泄物关了机。 </li><li>用干净的乙醇70％的底板。喷洒下来整机用二氧化氯消毒。 </li><li>天- 4 - 3， - 2，和- 1圆周率，预先设定的旋转杆与相应的训练协议参数，如表1所述，并重复步骤2.1.2至2.1.12。 </li><li>获得基线措施后，感染与TMEV老鼠。允许6天PI恢复期。 </li></ol></li></ol><table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td rowspan="2"> 协议 </td><td rowspan="2"> 测试日 </td><td rowspan="2"> 频率 </td><td rowspan="2"> 起动转速（rpm） </td><td rowspan="2"> 马克斯转速（rpm） </td><td> 促进 </td><td> 试验 </td><td> ITI </td></tr><tr><td> （转/秒） </td><td> 为（N×秒） </td><td> （分钟） </td></tr><tr><td rowspan="4"> 训练 </td><td> - 5 DPI </td><td> 1天</td><td> 1 </td><td> 12 </td><td> 01/03 </td><td> 3x240秒</td><td> 3 </td></tr><tr><td> - 4 DPI </td><td> 1天</td><td> 1 </td><td> 13 </td><td> 01/03 </td><td> 3x240秒</td><td> 3 </td></tr><tr><td> - 3 DPI </td><td> 1天</td><td> 1 </td><td> 14 </td><td> 01/03 </td><td> 3x240秒</td><td> 3 </td></tr><tr><td> - 2; - 1 DPI </td><td> 1天</td><td>五</td><td> 40 </td><td> 01/03 </td><td> 3x240秒</td><td> 3 </td></tr><tr><td rowspan="2"> 试验 </td><td>从+7至+50 DPI </td><td> 2 /周</td><td>五</td><td> 40 </td><td> 05/30 </td><td> 3x240秒</td><td> 3 </td></tr><tr><td>从+51到+150 DPI </td><td> 1周</td><td>五</td><td> 40 </td><td> 05/30 </td><td> 3x240秒</td><td> 3 </td></tr></tbody></table> 表1:转棒参数培训与实验方案。 <ol start="2"><li> 该转棒实验协议 <ol><li> +7 dpi的，预先设定的旋转杆与相应的实验协议参数，如表中所述1。重复步骤2.1.2至2.1.10。 </li><li>在审判＃3月底，权衡每个鼠标和记体重的数据表上。清洁和消毒旋转杆按步骤2.1.11和2.1.12。 </li><li>测试小鼠每周两次对下列6周，如上所述。 6周每周一次使用相同experimenta（其中小鼠可能已经达到了一个平台期）8,9，测试小鼠后l协议。实际测试协议持续150天的平均值，这取决于具体的疾病过程。 </li></ol></li><li> 神经功能指数 <ol><li>导出原始数据到电子表格文件，并分析结果。 </li><li>如运行时间（ 图3A）Express数据:这是在正常运行时间加上被动旋转时间减去旋转延迟时间（ 表2）10。计算每日的三次试验的平均运行时间。 </li><li>快递数据作为神经功能指数（NFI; 图3B）。 <ol><li>计算每个鼠标的基准性能门槛。基准性能阈值确定为从天所有的运行时间的平均值±15 + 45圆周6,7。 </li><li>计算NFI为最近的三个平均运行时间由特定的鼠标6,7的基准性能阈值除以平均值&lt;/ SUP&gt;。 注意:如果测试运行时间的日+ 72小鼠，+ 76和+ 79圆周率是55秒，45秒和50秒，基线时间为相同的鼠标为135秒时，NFI该鼠标在+79 dpi将是[（45 + 50 + 55）/ 3] / 135或0.37。 </li></ol></li><li>快递数据作为调整NFI（adjNFI; 图3C）:由单一实验中总体值调整NFI数据。 <ol><li>除以平均NFI由假治疗组在该特定日期获得的NFI值计算adjNFI。 </li></ol></li></ol></li></ol><table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> 术语 </td><td> 定义 </td></tr><tr><td>正常运行时间</td><td>的总时间的小鼠花费在转动杆主动运行， 即 ，等待时间下降。 </td></tr><tr><td>被动旋转时间</td><td>在一个时间安装鼠标一直在被动旋转模式杆。 </td></tr><tr><td>旋转延迟时间</td><td>被动旋转模式期间的时间的小鼠保持在杆的量</td></tr><tr><td>被动旋转模式</td><td>当鼠标抓住杆和无需走动旋转。 </td></tr><tr><td>总会话时间</td><td>时间总量鼠标保持在会议期间旋转杆。 </td></tr><tr><td>基准性能</td><td>预损伤电机性能评估，以确定该最低性能阈值。 </td></tr><tr><td>神经功能指数（NFI） </td><td>临床指标，其中每个小鼠电机性能进行比较， 即，运行时间，在任何时候其最佳性能。 </td></tr><tr><td>调整后的神经功能指数（adjNFI） </td><td>当施加正常化过程通过为人口值来调节NFI数据单一的实验。 </td></tr><tr><td>总体值</td><td>在特定的一天假治疗组获得的平均NFI值。 </td></tr></tbody></table> 表2:采用量化神经损伤转棒参数的定义。

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The authors have nothing to disclose.

尽管有一些限制，所述旋转杆的性能测试代表用于评估TMEV-IDD运动功能和功能障碍，以及药理干预对残疾进展中的小鼠的影响的重要工具。 旋转试验是在1957年首次描述为在啮齿类动物11测量神经功能缺损的工具。鼠害必须走在旋转的杆，随着转速，并尽量避免坠落到地面。落入延迟被记录并用作定量终点运动功能:越神经功能缺损的动物有，越早它们脱落杆。此测试在...

Theiler鼠脑脊髓炎病毒（TMEV）是一种嗜神经单链RNA病毒持续感染的鼠的中枢神经系统（CNS）。在易感小鼠中，感染TMEV导致免疫介导的慢性进行性脱髓鞘疾病，称为TMEV诱导脱髓鞘疾病（TMEV-IDD）。小鼠的实验性感染发生的疾病过程类似，在多发性硬化症（MS）的渐进形式看到。 TMEV-IDD的特征在于两个不同的阶段:急性期和慢性期。急性期是一种温和的，通常亚临床性脑炎1,2。第二，慢性期，开始约一个月感染后，由一个缓慢进展残疾特征在于脱髓鞘，炎症和轴索损害1,2。在小鼠中观察到的疲软与痉挛，偶尔严重的强直性痉挛有关。 由于目前还没有medicatioNS改善逐步残疾患者，研究人员特别是TMEV，国际直拨电话，它代表了监测疾病修饰药物对疾病进展的影响，最佳的动物模型所吸引。然而，在小鼠中，以及在MS患者，残疾进展的监测需要在延长的时间周期的连续的临床观察。在小鼠中，对于残疾进展的长期监测可与旋转杆的性能测试来完成。 在转棒性能测试是一种标准的行为测试，用于评估运动相关的功能，例如协调性，平衡，和疲劳在啮齿类动物。小鼠必须保持平衡的转弯杆，这是在连续加速旋转;落入时间延迟从该杆被记录。与神经功能障碍的动物不能长留在旋转杆作为对照，他们当转速超过正常脱落的电机容量。更多的神经功能缺损动物们，他们越早脱落杆，以及更短的时间延迟。 旋转试验在传统的视觉评分系统的优点在于，它产生一个目标，可测量的可变时间延迟该最终可用于统计分析以量化疗法和实验程序3的效果。 在免疫学（LONI）中的达特茅斯实验室，将小鼠进行的适配协议，在那里它们被TMEV感染前测试，以便与机器使他们熟悉并评估其正常的"基线"平衡协调和电机控制4， 5。一旦基线建立和小鼠被感染TMEV，它们过了一段数月测定一次或每周两次。实际测试协议为期150天的平均值，从而允许进行评估平衡，协调和电机控制的降幅比脱髓鞘疾病的全过程。 几百TMEV，IDD和假治疗的小鼠迄今已测试达特茅斯神经功能障碍。这些小鼠接受了各种免疫调节治疗方法，但没有药剂已发现可有效地改善残疾进展6,7。本文章及相关协议描述了如何通过定性TMEV-IDD小鼠显示进行性神经损害。特别是，该协议提供的认为是一般适合学习使用旋转试验TMEV-IDD小鼠神经失具体的测试参数的建议。此过程提供对照评估一个基线（1）该小鼠模型来进行性MS和（2）它的实用性，用于测试旨在治疗进行性神经疾病如MS疗法的相关性。显然，转棒性能测试和当前优化测试参数和协议不仅在该TMEV-IDD小鼠模型检测进行性神经障碍是有用的，但是，可以在中神经变性疾病的其他病毒诱导的和/或遗传小鼠模型揭示损伤是有用的。

<table><tbody><tr><td>Mice SJL/JCrHsd 4 to 6 weeks old</td><td>Envigo</td><td>#052</td><td></td></tr><tr><td>TMEV virus stock</td><td></td><td></td><td></td></tr><tr><td>Isoflurane vaporizer</td><td>Harvard Apparatus</td><td>#340471</td><td></td></tr><tr><td>Insulin Syringes U- 100 29 g x 0.5 cc</td><td>BD</td><td>#328203</td><td></td></tr><tr><td>Rotamex-5 4 Lane Rota-Rod for Mice with RS-232 and Software</td><td>Columbus Instruments</td><td>#0890M</td><td></td></tr></tbody></table>

measuring progressive neurological disability in a mouse model of multiple sclerosis

After intracerebral infection with the Theiler's Murine Encephalomyelitis Virus (TMEV), susceptible SJL mice develop a chronic-progressive demyelinating disease, with clinical features similar to the progressive forms of multiple sclerosis (MS). The mice show progressive disability with loss of motor and sensory functions, which can be assessed with multiple apparatuses and protocols. Among them, the Rotarod performance test is a very common behavioral test, its advantage being that it provides objective measurements, but it is often used assuming that it is straightforward and simple. In contrast to visual scoring systems used in some models of MS, which are highly subjective, the Rotarod test generates an objective, measurable, continuous variable (i.e., length of time), allowing almost perfect inter-rater concordances. However, inter-laboratory reliability is only achieved if the various testing parameters are replicated. In this manuscript, recommendations of specific testing parameters, such as size, speed, and acceleration of the rod; amount of training given to the animals; and data processing, are presented for the Rotarod test.

该代表性实验的目的是比较由丹尼尔斯（DA）菌株和TMEV豆应变诱导的神经学障碍。本研究的目的，一组32例女性SJL小鼠用TMEV感染脑内，无论是DA株（N = 16）或豆株（N = 16），其临床症状随着时间的推移进行了监测。 20只小鼠的另外一个组假处理（ 即生理盐水溶液注入脑内），并担任健康对照组。 在转棒性能试验来评价?...

Watch this Scientific Journal Video about Measuring Progressive Neurological Disability in a Mouse Model of Multiple Sclerosis at JoVE.com

Measuring Progressive Neurological Disability in a Mouse Model of Multiple Sclerosis

An optimized testing protocol is presented in this paper for the Rotarod performance test, used for measuring progressive neurological disability in TMEV-infected mice.

在多发性硬化症的小鼠模型测量进行性神经功能残疾

After intracerebral infection with the Theiler's Murine Encephalomyelitis Virus (TMEV), susceptible SJL mice develop a chronic-progressive demyelinating ...

measuring-progressive-neurological-disability-mouse-model-multiple

Research

JoVE Journal

Neuroscience

10.9K 次观看.  Geisel School of Medicine at Dartmouth. An optimized testing protocol is presented in this paper for the Rotarod performance test, used for measuring progressive neurological disability in TMEV-infected mice. 

视频: Measuring Progressive Neurological Disability in a Mouse Model of Multiple Sclerosis

Measuring Motor Coordination in Mice

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such as emotionality or cognition can be assessed, however, it is vital to determine whether the motor capabilities of e.g. a mutant or lesioned mouse allow such an assessment. Performance on a maze task requiring strength and coordination, such as the Morris water maze, might well be impaired in a mouse by motor, rather than cognitive, impairments, so it is essential to selectively dissect the latter from the former. For example, sensorimotor impairments caused by NMDA antagonists have been shown to impair water maze performance2. 
Motor coordination has traditionally been assessed in mice and rats by the rotarod test, in which the animal is placed on a horizontal rod that rotates about its long axis; the animal must walk forwards to remain upright and not fall off. Both set speed and accelerating versions of the rotarod are available.
The other three tests described in this article (horizontal bar, static rods and parallel bars) all measure coordination on static apparatus. The horizontal bar also requires strength for adequate performance, particularly of the forelimbs as the mouse initially grips the bar just with the front paws. 
Adult rats do not perform well on tests such as the static rods and parallel bars (personal observations); they appear less well coordinated than mice. I have only tested male rats, however, and male mice seem generally less well coordinated than females. Mice appear to have a higher strength:weight ratio than rats; the Latin name, Mus musculus, seems entirely appropriate. The rotarod, the variations of the foot fault test12 or the Catwalk (Noldus)15 apparatus are generally used to assess motor coordination in rats.

Protocols are presented for two established motor coordination tasks, the accelerating rotarod and horizontal bar, also two tests developed in Oxford recently, the static rods and parallel bars. These tests can detect motor impairments potentially of interest in their own right, as well as being possible variables in tests of other areas of behavior.

测量小鼠运动协调

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such ...

科研

神经科学

Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis (ALS)

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder resulting in progressive degeneration of motoneurons. Peak of onset is around 60 years for the sporadic disease and around 50 years for the familial disease. Due to its progressive course, 50% of the patients die within 30 months of symptom onset. In order to evaluate novel treatment options for this disease, genetic mouse models of ALS have been generated based on human familial mutations in the SOD gene, such as the SOD1 (G93A) mutation. Most important aspects that have to be evaluated in the model are overall survival, clinical course and motor function. Here, we demonstrate the clinical evaluation, show the conduction of two behavioural motor tests and provide quantitative scoring systems for all parameters. Because an in depth analysis of the ALS mouse model usually requires an immunohistochemical examination of the spinal cord, we demonstrate its preparation in detail applying the dorsal laminectomy method. Exemplary histological findings are demonstrated. The comprehensive application of the depicted examination methods in studies on the mouse model of ALS will enable the researcher to reliably test future therapeutic options which can provide a basis for later human clinical trials.

Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis ALS

A mouse model for amyotrophic lateral sclerosis (ALS) is examined clinically and behaviorally. As a prerequisite for an accompanying immunohistological analysis the preparation of the spinal cord is depicted in detail.

临床试验和脊髓去除在肌萎缩性侧索硬化症的小鼠模型（ALS）

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder resulting in progressive degeneration of motoneurons. Peak of onset is around 60 ...

医学

A Quick Phenotypic Neurological Scoring System for Evaluating Disease Progression in the SOD1-G93A Mouse Model of ALS

The SOD1-G93A transgenic mouse is the most widely used animal model of amyotrophic lateral sclerosis (ALS). At ALS TDI we developed a phenotypic screening protocol, demonstrated in video herein, which reliably assesses the neuromuscular function of SOD1-G93A mice in a quick manner. This protocol encompasses a simple neurological scoring system (NeuroScore) designed to assess hindlimb function. NeuroScore is focused on hindlimb function because hindlimb deficits are the earliest reported neurological sign of disease in SOD1-G93A mice. The protocol developed by ALS TDI provides an unbiased assessment of onset of paresis (slight or partial paralysis), progression and severity of paralysis and it is sensitive enough to identify drug-induced changes in disease progression. In this report, the combination of a detailed manuscript with video minimizes scoring ambiguities and inter-experimenter variability thus allowing for the protocol to be adopted by other laboratories and enabling comparisons between studies taking place at different settings. We believe that this video protocol can serve as an excellent training tool for present and future ALS researchers.

This video protocol describes a sensitive, reliable, and quick method for evaluating the neuromuscular deficits in a transgenic mouse model of amyotrophic lateral sclerosis.

一个快速的表型神经评分系统在ALS的SOD1-G93A小鼠模型评估疾病进展

The SOD1-G93A transgenic mouse is the most widely used animal model of amyotrophic lateral sclerosis (ALS). At ALS TDI we developed a phenotypic screening ...

Sagittal Plane Kinematic Gait Analysis in C57BL/6 Mice Subjected to MOG35-55 Induced Experimental Autoimmune Encephalomyelitis

Kinematic gait analysis in the sagittal plane has frequently been used to characterize motor deficits in multiple sclerosis (MS). We describe the application of these techniques to identify gait deficits in a mouse model of MS, known as experimental autoimmune encephalomyelitis (EAE). Paralysis and motor deficits in mice subjected to EAE are typically assessed using a clinical scoring scale. However, this scale yields only ordinal data that provides little information about the precise nature of the motor deficits. EAE disease severity has also been assessed by rotarod performance, which provides a measure of general motor coordination. By contrast, kinematic gait analysis of the hind limb in the sagittal plane generates highly precise information about how movement is impaired. To perform this procedure, reflective markers are placed on a hind limb to detect joint movement while a mouse is walking on a treadmill. Motion analysis software is used to measure movement of the markers during walking. Kinematic gait parameters are then derived from the resultant data. We show how these gait parameters can be used to quantify impaired movements of the hip, knee, and ankle joints in EAE. These techniques may be used to better understand disease mechanisms and identify potential treatments for MS and other neurodegenerative disorders that impair mobility.

Kinematic gait analysis in the sagittal plane yields highly precise information about how movement is executed. We describe the application of these techniques to identify gait deficits for mice subjected to autoimmune-mediated demyelination. These methods may also be used to characterize gait deficits for other mouse models featuring impaired locomotion.

MOG35-55 诱发自身免疫性脑脊髓炎 C57BL/6 小鼠矢状面运动步态分析

Kinematic gait analysis in the sagittal plane has frequently been used to characterize motor deficits in multiple sclerosis (MS). We describe the ...

Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Mice

Experimental autoimmune encephalomyelitis (EAE) in SJL/J mice is a model for relapsing-remitting multiple sclerosis (RRMS). Clinical EAE scores describing motor function deficits are basic readouts of the immune-mediated inflammation of the spinal cord. However, scores and body weight do not allow for an in vivo assessment of brain inflammation and optic neuritis. The latter is an early and frequent manifestation in about 2/3 of MS patients. Here, we show methods for bioluminescence and near-infrared live imaging to assess EAE evoked optic neuritis, brain inflammation, and blood-brain barrier (BBB) disruption in living mice using an in vivo imaging system. A bioluminescent substrate activated by oxidases primarily showed optic neuritis. The signal was specific and allowed the visualization of medication effects and disease time courses, which paralleled the clinical scores. Pegylated fluorescent nanoparticles that remained within the vasculature for extended periods of time were used to assess the BBB integrity. Near-infrared imaging revealed a BBB leak at the peak of the disease. The signal was the strongest around the eyes. A near-infrared substrate for matrix metalloproteinases was used to assess EAE-evoked inflammation. Auto-fluorescence interfered with the signal, requiring spectral unmixing for quantification. Overall, bioluminescence imaging was a reliable method to assess EAE-associated optic neuritis and medication effects and was superior to the near-infrared techniques in terms of signal specificity, robustness, ease of quantification, and cost.

We show a technique for in vivo live bioluminescence and near-infrared imaging of optic neuritis and encephalitis in the experimental autoimmune encephalomyelitis (EAE) model for multiple sclerosis in SJL/J mice.

多发性硬化症的小鼠模型EAE视神经炎及脑部炎症的生物发光和近红外成像

Experimental autoimmune encephalomyelitis (EAE) in SJL/J mice is a model for relapsing-remitting multiple sclerosis (RRMS). Clinical EAE scores describing ...

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 ...

脊髓组织学染色用于评估 EAE 疾病严重程度

Scoring Central Nervous System Inflammation, Demyelination, and Axon Injury in Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) is a common immune-based model of multiple sclerosis (MS). This disease can be induced in rodents by active immunization with protein components of the myelin sheath and Complete Freund&#39;s adjuvant (CFA) or by the transfer of myelin-specific T effector cells from rodents primed with myelin protein/CFA into na&#239;ve rodents. The severity of EAE is typically scored on a 5-point clinical scale that measures the degree of ascending paralysis, but this scale is not optimal for assessing the extent of recovery from EAE. For example, clinical scores remain high in some EAE models (e.g., myelin oligodendrocyte glycoprotein [MOG] peptide-induced model of EAE) despite the resolution of inflammation. Thus, it is important to complement clinical scoring with histological scoring of EAE, which also provides a means to study the underlying mechanisms of cellular injury in the central nervous system (CNS).
Here, a simple protocol is presented to prepare and stain spinal cord and brain sections from mice and to score inflammation, demyelination, and axonal injury in the spinal cord. The method for scoring leukocyte infiltration in the spinal cord can also be applied to score brain inflammation in EAE. A protocol for measuring soluble neurofilament light (sNF-L) in the serum of mice using a Small Molecule Assay (SIMOA) assay is also described, which provides&#160;feedback on the extent of overall CNS injury in live mice.

作者聚焦：揭示多发性硬化症中肥胖与自身免疫性炎症之间的通路 

Experimental autoimmune encephalomyelitis (EAE) serves as an animal model of multiple sclerosis. This article describes an approach for scoring spinal cord inflammation, demyelination, and axonal injury in EAE. Additionally, a method to quantify soluble neurofilament light levels in the mice serum is presented, facilitating the assessment of axonal injury in live mice.

对实验性自身免疫性脑脊髓炎中的中枢神经系统炎症、脱髓鞘和轴突损伤进行评分

Experimental autoimmune encephalomyelitis (EAE) is a common immune-based model of multiple sclerosis (MS). This disease can be induced in rodents by ...

免疫与感染

使用 MouseWalker 量化脊髓损伤小鼠的运动功能障碍

Using the MouseWalker to Quantify Locomotor Dysfunction in a Mouse Model of Spinal Cord Injury

The execution of complex and highly coordinated motor programs, such as walking and running, is dependent on the rhythmic activation of spinal and supra-spinal circuits. After a thoracic spinal cord injury, communication with upstream circuits is impaired. This, in turn, leads to a loss of coordination, with limited recovery potential. Hence, to better evaluate the degree of recovery after the administration of drugs or therapies, there is a necessity for new, more detailed, and accurate tools to quantify gait, limb coordination, and other fine aspects of locomotor behavior in animal models of spinal cord injury. Several assays have been developed over the years to quantitatively assess free-walking behavior in rodents; however, they usually lack direct measurements related to stepping gait strategies, footprint patterns, and coordination. To address these shortcomings, an updated version of the MouseWalker, which combines a frustrated total internal reflection (fTIR) walkway with tracking and quantification software, is provided. This open-source system has been adapted to extract several graphical outputs and kinematic parameters, and a set of post-quantification tools can be to analyze the output data provided. This manuscript also demonstrates how this method, allied with already established behavioral tests, quantitatively describes locomotor deficits following spinal cord injury.

作者聚焦:使用 MouseWalker 量化脊髓损伤小鼠模型中的运动功能障碍  

An experimental pipeline to quantitatively describe the locomotor pattern of freely walking mice using the MouseWalker (MW) toolbox is provided, ranging from initial video recordings and tracking to post-quantification analysis. A spinal cord contusion injury model in mice is employed to demonstrate the usefulness of the MW system.

在多发性硬化症的小鼠模型测量进行性神经功能残疾

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此视频中的章节

测量小鼠运动协调

临床试验和脊髓去除在肌萎缩性侧索硬化症的小鼠模型（ALS）

一个快速的表型神经评分系统在ALS的SOD1-G93A小鼠模型评估疾病进展

MOG35-55 诱发自身免疫性脑脊髓炎 C57BL/6 小鼠矢状面运动步态分析

多发性硬化症的小鼠模型EAE视神经炎及脑部炎症的生物发光和近红外成像

体内运动神经元变性小鼠前肢肌作用电位的电生理测量

对实验性自身免疫性脑脊髓炎中的中枢神经系统炎症、脱髓鞘和轴突损伤进行评分

对实验性自身免疫性脑脊髓炎中的中枢神经系统炎症、脱髓鞘和轴突损伤进行评分

对实验性自身免疫性脑脊髓炎中的中枢神经系统炎症、脱髓鞘和轴突损伤进行评分

使用MouseWalker量化脊髓损伤小鼠模型中的运动功能障碍