Organotypic culture assay for neuromuscular synaptic degeneration and function

Kosala N. Dissanayake, Robert Chang Chih Chou, Rosalind Brown, Richard R. Ribchester*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

We describe here an organotypic culture system we have used to investigate mechanisms that maintain structure and function of axon terminals at the neuromuscular junction (NMJ). We developed this by taking advantage of the slow Wallerian degeneration phenotype in mutant Wlds mice, using these to compare preservation of NMJs with degeneration in nerve-muscle preparations from wild-type mice. We take hind limb tibial nerve/flexor digitorum brevis and lumbrical muscles and incubate them in mammalian physiological saline at 32 °C for 24–48 h. Integrity of NMJs can then be compared using a combination of electrophysiological and morphological techniques. We illustrate our method with data showing synaptic preservation ex vivo in nerve-muscle explants from Sarm-1 null-mutant mice. The ex vivo assays of NMJ integrity we describe here may therefore be useful for detailed investigation of synaptic maintenance and degeneration.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages145-157
Number of pages13
DOIs
Publication statusPublished - 11 Jun 2020

Publication series

NameMethods in Molecular Biology
Volume2143
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords / Materials (for Non-textual outputs)

  • Electrophysiology
  • Ex vivo assay
  • Immunostaining
  • Neuromuscular junction
  • Wallerian degeneration

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