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1. Adult rat skeletal muscles were dissociated by collagenase treatment and trituration, and the isolated muscle fibres were maintained in vitro for 2-3 weeks. At various stages, the fibres were examined physiologically and morphologically. 2. The isolated fibres underwent some changes characteristic of muscle denervated in vivo. For instance, input resistance increased and extrajunctional acetylcholine (ACh) receptors appeared. In addition, the beginning stages of apparent muscle fibre fragmentation were observed. 3. In other respects, the cultured isolated fibres behaved differently than in vivo denervated fibres. Fibrillation developed only occasionally in vitro. The onset of ACh supersensitivity was slower (6 days) than after denervation in vivo (2-3 days). Some fibres developed localized regions of destriation, which apparently was due to loss of in-register alignment of myofibrils.
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