Organotypic Culture Assay for Neuromuscular Synaptic Degeneration and Function.
| Autor: | Dissanayake KN; Centre for Discovery Brain Sciences and the Euan MacDonald Centre for Motor Neurone Disease Research, University of Edinburgh, Edinburgh, UK., Chou RC; Centre for Discovery Brain Sciences and the Euan MacDonald Centre for Motor Neurone Disease Research, University of Edinburgh, Edinburgh, UK., Brown R; Centre for Discovery Brain Sciences and the Euan MacDonald Centre for Motor Neurone Disease Research, University of Edinburgh, Edinburgh, UK., Ribchester RR; Centre for Discovery Brain Sciences and the Euan MacDonald Centre for Motor Neurone Disease Research, University of Edinburgh, Edinburgh, UK. Richard.Ribchester@ed.ac.uk. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2020; Vol. 2143, pp. 145-157. |
| DOI: | 10.1007/978-1-0716-0585-1_11 |
| Abstrakt: | 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 Wld s 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. |
| Databáze: | MEDLINE |
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