Scanning and freeze-fracture study of larval nerves and neuromuscular junctions in Manduca sexta.

The nerves and nerve terminals to tonic larval muscle fibers in third and fifth instar caterpillars were studied to compare them with those formed by the same motor neurons on phasic flight muscles in adult moths. Scanning micrographs showed a primary nerve branch running the length of each fiber, with secondary nerve branches extending from it at intervals. There was a great deal of variability in both the length of the branches and the distance from the nerve at which the neuromuscular junctions were formed. The rapid increase in muscle fiber size during larval development may be responsible for this variability. The nerves and junctions were often found to be obscure by overlying fibroblasts and tracheoblasts or entering the deep muscle clefts. Those examined were similar in appearance to the adult junctions formed by the same neurons, although some may have formed single branches instead of y-shapes. The membrane specializations of the synapse seen in freeze-fractured specimens were similar to those of the adult junction. However, the overall shape of the nerve terminal within the junction differed. The larval nerve terminals appeared varicose instead of having a uniform diameter. The spacing of the nerve plaques varied, in contrast with the relatively straight alignment and even spacing of plaques found in adult junctions. Such differences could result from an interaction between the motor neuron and the two different types of muscle fiber that it innervates, an intrinsic change in the motor neurons themselves that occurs with metamorphosis, or a plastic functional response that occurs as a result of the different types of motor patterns that are used in the two stages.