Synapse formation between dissociated nerve and muscle cells in low density cell cultures.

Some of the neurons dissociated from embryonic chick spinal cord and maintained in low density cell cultures establish functional contacts with muscle fibers that had formed in vitro from previously plated myoblasts. Exposure of young muscle cultures to d-arabinofuranosylcytosine and subsequent maintenance of combined neuron-muscle cultures in low embryo extract media eliminated most of the peripheral and central supporting cells. The synapses are cholinergic, and transmitter release which, even in the youngest cultures is quantal, is regulated by Ca2+ and Mg2+. The major functional differences from adult vertebrate neuromuscular junctions are: slow synaptic potential time course, low end-plate potential quantum content and miniature end-plate potential frequency, occurrence of spontaneous end-plate potentials, and lack of functional acetylcholinesterase. In addition, direct electrical coupling between nerve and muscle has been observed in four cases.

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