Observation of Acetylcholine Receptor Aggregation at In Vitro Chick Nerve/Muscle Contracts
Forsch, Randall T.
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The early events in the in vitro formation of the neuromuscular junction have yet to be entirely revealed. Using an in vitro chick embryonic ciliary ganglion neuron/skeletal muscle system, experiments were undertaken to determine the time-course for the appearance of AChR clustering, the location of these clusters, and the role of extra-junctional and junctional AChR in this early clustering. Neuron and muscle tissue culture, AChR marking with fluorescently-labelled α-bungarotoxin and anti-AChR monoclonal anti-bodies, and time-lapse cinemicroscopy were used extensively. It was determined that AChR aggregation probably occurs within 3 to 4 hours after contact with the neuron's soma and/or growth cone. The AChR involved appear to be extra-junctional.