Sleigh James N, Burgess Robert W, Gillingwater Thomas H, Cader M Zameel
Nuffield Department of Clinical Neurosciences, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DU, UK; The Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK.
The Jackson Laboratory, Bar Harbor, ME 04609, USA.
J Neurosci Methods. 2014 Apr 30;227:159-65. doi: 10.1016/j.jneumeth.2014.02.005. Epub 2014 Feb 14.
The neuromuscular junction (NMJ) is a specialised synapse formed between a lower motor neuron and a skeletal muscle fibre, and is an early pathological target in numerous nervous system disorders, including amyotrophic lateral sclerosis (ALS), Charcot-Marie-Tooth disease (CMT), and spinal muscular atrophy (SMA). Being able to accurately visualise and quantitatively characterise the NMJ in rodent models of neurological conditions, particularly during the early stages of disease, is thus of clear importance.
We present a method for dissection of rodent deep lumbrical muscles located in the hind-paw, and describe how to perform immunofluorescent morphological analysis of their NMJs.
These techniques allow the temporal assessment of a number of developmental and pathological NMJ phenotypes in lumbrical muscles.
Small muscles, such as the distal hind-limb lumbrical muscles, possess a major advantage over larger muscles, such as gastrocnemius, in that they can be whole-mounted and the entire innervation pattern visualised. This reduces preparation time and ambiguity when evaluating important neuromuscular phenotypes.
Together, these methods will allow the reader to perform a detailed and accurate analysis of the neuromuscular system in rodent models of disease in order to identify pertinent features of neuropathology.
神经肌肉接头（NMJ）是在下运动神经元和骨骼肌纤维之间形成的一种特殊突触，并且是包括肌萎缩侧索硬化症（ALS）、夏科-马里-图斯病（CMT）和脊髓性肌萎缩症（SMA）在内的众多神经系统疾病的早期病理靶点。因此，能够在神经疾病的啮齿动物模型中准确可视化并定量表征神经肌肉接头，尤其是在疾病的早期阶段，显然具有重要意义。
我们提出了一种用于解剖位于后爪的啮齿动物深部蚓状肌的方法，并描述了如何对其神经肌肉接头进行免疫荧光形态学分析。
这些技术能够对蚓状肌中多种发育和病理性神经肌肉接头表型进行时间评估。
诸如后肢远端蚓状肌这样的小肌肉，相较于诸如腓肠肌这样的大肌肉具有一个主要优势，即它们可以进行整装处理并可视化整个神经支配模式。这在评估重要的神经肌肉表型时减少了准备时间和不确定性。
总之，这些方法将使读者能够在疾病的啮齿动物模型中对神经肌肉系统进行详细而准确的分析，以便识别神经病理学的相关特征。
