Differences in the localization of the postsynaptic nitric oxide synthase I and acetylcholinesterase suggest a heterogeneity of neuromuscular junctions in rat and mouse skeletal muscles

Abstract

Recently, nitric oxide synthase (NOS) I has been identified in skeletal muscle fibers, where the enzyme is found to be associated to the sarcolemma by the alpha 1-syntrophin-dystrophin complex. It has, however, been proposed that a substantial proportion of NOS I at the neuromuscular junction (NMJ) is of neuronal origin. We have, therefore, investigated the distribution of NOS I in NMJ of normal rats and mice as well as mdx mice which lack dystrophin and, consequently, NOS I in the sarcolemma region by enzyme histochemical and immunohistochemical techniques. Sites of NOS I accumulation, evident at NMJ of healthy animals, were absent in mdx mice, indicating a predominantly, if not exclusively, postsynaptic localization of NOS I at NMJ. Moreover, simultaneous demonstration of acetylcholinesterase (AChE) activity revealed a heterogeneity of NMJ in rat and mouse skeletal muscles: type I showed only AChE activity and was found to predominate; type II was spatially separated from the AChE-positive NMJ, occurred less frequently and contained both AChE activity and NOS I. These data suggest that type II NMJ are provided with additional regulatory mechanisms, such as free radical signaling by the NOS I-derived NO which may exert modulatory effects on the choline acetyltransferase/ACh/AChE pathway. Furthermore, type II may represent those NMJ where recently glutamate-gated NMDA-type Ca2+ channels have been described, which in analogy to those in the nervous system may serve also in skeletal muscle fibers as NOS I activators.