Light and electron microscopic identification of nerve terminal sprouting and retraction in normal adult frog muscle

A combined light and electron microscopic study was performed on neuromuscular junctions of normal adult frogs. In a previous investigation signs of new synapse formation, as well as abandoned former synaptic sites, have been observed in normal muscles (Wernig, Pécot-Dechavassine & Stöver, 1980a, b). Here we performed a detailed light and electron microscopic correlation to investigate those parts of junctions which, after staining for cholinesterase (ChE) and presynaptic axon terminals, were suspected either to be newly formed or sites abandoned by the presynaptic nerve and the Schwann cell. Thin presynaptic nerve branches, enclosed by Schwann cell sheaths along most of their length, formed synaptic contacts with the muscle fibre only at small circumscribed areas. In these regions post-synaptic secondary folds (invariably present at mature synapses) were either missing or were less well developed. At these small contacts, binding sites for fluorescein-labelled alpha-bungarotoxin were usually present. At other sites the ChE reaction product was present but an axon could not be detected in silver-stained preparations. Electron microscopic observation revealed post-synaptic secondary folds filled with ChE reaction product while the presynaptic axon and Schwann cell were missing. The sites with ChE remnants can thus be regarded as abandoned former synaptic contacts. No binding of fluorescein-labelled alpha-bungarotoxin could be detected at such sites. These findings confirm earlier suggestions that synaptic contacts in frog muscle are normally undergoing continual remodelling. The lack of binding sites for fluorescein-labelled alpha-bungarotoxin at abandoned synaptic sites suggests that a neural or Schwann cell factor is important for the maintainance of synaptic acetylcholine receptors.