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.
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Acknowledgments
Work in our laboratory is supported by grants from the Medical Research Council (MR/M024075/1), the Motor Neurone Disease Association (838-791), and the Euan MacDonald Centre for Motor Neurone Disease Research. We thank Professor Michael P Coleman and Dr. Jon Gilley for providing tissue from Sarm-1 null-mutant mice.
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Dissanayake, K.N., Chou, R.CC., Brown, R., Ribchester, R.R. (2020). Organotypic Culture Assay for Neuromuscular Synaptic Degeneration and Function. In: Babetto, E. (eds) Axon Degeneration. Methods in Molecular Biology, vol 2143. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0585-1_11
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DOI: https://doi.org/10.1007/978-1-0716-0585-1_11
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