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Muscle derived cells, mainly myoblastes and fibroblasts are cultured in monolayers. Immediately after myoblaste fusion, whole transverse slices of 13-day-old rat embryos spinal cords with dorsal root ganglia attached are placed on the muscle monolayer. After 24h neuritis are observed growing out of the spinal cord explants. They make contacts with myotubes and induce the first contractions. Quickly thereafter, innervated muscle fibres located in proximity to the spinal cord explants, are virtually continuously contracting. Innervated fibres are morphologically and spatially distinct from the non-innervated ones and could easily be distinguished from them (Askanas et al., 1987).
The ability of a molecule to promote spinal motor neuron survival, neurites outgrowth and/or neurons functionality can be evaluated by measuring neurite sprouting (number and length), innervation rate (number of spinal cord-muscle couple which lead to an innervation), innervation area (surface covered by innervated muscle fibers per spinal cord explant), ACh receptor aggregation as well as several other parameters depending upon the mode of action of the compound (Braun et al., 1996).
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Askanas V., Kwan H., Alvarez R.B., Engel W.K., Kobayashi T., Martinuzzi A. and Hawkins E.F. (1987). De novo neuromuscular junction formation on human muscle fibres cultured in monolayer and innervated by foetal rat spinal cord: ultrastructural and ultrastructural-cytochemical studies. 