Neuromuscular junction development in the cutaneous pectoris muscle of Rana catesbeiana

Synaptic specializations were studied in the developing cutaneous pectoris muscle of Rana catesbeiana tadpoles and froglets to correlate nerve terminal morphology (by light and electron microscopy), accumulation of acetylcholine receptors, and the ability of the muscle to contract following nerve stimulation. This correlated approach was used to determine the developmental timing and possible causal relationship of events in nerve and muscle maturation at the neuromuscular junction. Initially, the cutaneous pectoris nerve trunk was present in the undifferentiated presumptive cutaneous pectoris mesenchyme, prior to muscle maturation. At stage XII when the muscle was first able to contract weakly in response to nerve stimulation, the motor nerve terminal endings were simple bulbous enlargements associated with diffuse subneural aggregations of acetylcholine receptors (indicated by diffuse speckles of rhodamine alpha-bungarotoxin fluorescence). Before stage XII no rhodamine alpha-bungarotoxin fluorescence was present anywhere in the muscle. The first stage in the organization of acetylcholine receptors at the neuromuscular junction was the accumulation of diffuse speckles of fluorescence beneath the terminal enlargements. This was followed by the clustering of receptors into small polygonal areas at each synaptic site, and finally the organization of receptors into parallel linear rows. Presumably this final stage was associated with formation of junctional folds. By stage XV the synapses were multiply innervated and had developed acetylcholinesterase activity. The general nerve terminal morphology and pattern of accumulation of acetylcholine receptors at cutaneous pectoris neuromuscular junctions were similar to those of the adult throughout metamorphic climax except that they still contained more than one motor axon. After metamorphic climax, elimination of multiple innervation occurred.