Dissection of the transversus abdominis muscle for whole-mount neuromuscular junction analysis.

Dissection of the transversus abdominis muscle for whole-mount neuromuscular junction analysis.
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DOI:
10.3791/51162
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发表时间:
2014-01-11
期刊:
Journal of visualized experiments : JoVE
影响因子:
--
通讯作者:
Kothary R
Kothary R
中科院分区:
其他
文献类型:
--
作者:
Murray L;Gillingwater TH;Kothary R

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神经肌肉接头形态分析可以对给定运动神经元的生理状态提供重要的了解。与传统使用的较厚肌肉(例如后肢肌肉(例如腓肠肌))相比,对薄扁平肌肉的分析具有显着优势。薄肌肉可以全面概述给定肌肉的整个神经支配模式,从而可以选择性地识别易受攻击的运动神经元池。这些肌肉还可以分析运动单位大小、轴突分支和末端/节点发芽等参数。使用这些肌肉的一个常见障碍是获得解剖它们的技术专业知识。在本视频中,我们详细介绍了解剖幼鼠腹横肌 (TVA) 肌肉并进行免疫荧光以可视化轴突和神经肌肉接头 (NMJ) 的方案。我们证明该技术可以全面概述 TVA 肌肉的神经支配模式,并可用于研究儿童运动神经元疾病(脊髓性肌萎缩症)小鼠模型的 NMJ 病理学。
Analysis of neuromuscular junction morphology can give important insight into the physiological status of a given motor neuron. Analysis of thin flat muscles can offer significant advantage over traditionally used thicker muscles, such as those from the hind limb (e.g. gastrocnemius). Thin muscles allow for comprehensive overview of the entire innervation pattern for a given muscle, which in turn permits identification of selectively vulnerable pools of motor neurons. These muscles also allow analysis of parameters such as motor unit size, axonal branching, and terminal/nodal sprouting. A common obstacle in using such muscles is gaining the technical expertise to dissect them. In this video, we detail the protocol for dissecting the transversus abdominis (TVA) muscle from young mice and performing immunofluorescence to visualize axons and neuromuscular junctions (NMJs). We demonstrate that this technique gives a complete overview of the innervation pattern of the TVA muscle and can be used to investigate NMJ pathology in a mouse model of the childhood motor neuron disease, spinal muscular atrophy.
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