Terminal nerve sprouting at the frog neuromuscular junction induced by prolonged tetrodotoxin blockade of nerve conduction.

Abstract

The effects of a prolonged blockade of nerve conduction by tetrodotoxin on frog motor innervation were studied in the cutaneous pectoris muscle of Rana esculenta. Prolonged nerve blockade (up to 22 days) was obtained by repeated subperineural injections of tetrodotoxin. Changes in morphological parameters of neuromuscular junctions were investigated in muscles after staining with a combined cholinesterase-silver method. In addition, changes in the incidence of polyneuronal innervation were investigated conjointly by electrophysiology and morphology. Morphometric analysis of singly innervated muscle fibres of 60 microns diameter revealed insignificant changes during the first week of tetrodotoxin-nerve blockade. After 15 days of paralysis, the mean length of synaptic contacts and the mean length of terminal arborization per synapse were significantly increased as compared to controls (contralateral muscles and citrate buffer-injected controls). After 20-22 days, differences in synaptic and aborization mean lengths were accentuated and reached 44 and 43%, respectively. At that time, the mean number of terminal branching points and of continuous synaptic contacts were also significantly increased (around 20 and 50%, respectively). No changes in the length of abandoned gutters were observed. The incidence of focal polyaxonal innervation (detected morphologically) and of polyneuronal innervation (determined electrophysiologically) was unchanged. The results show that prolonged tetrodotoxin blockade induces sprouting of the terminal arborization which results in an extension of pre-existing nerve terminals and an increase in the complexity of terminal arborization by addition of new branches. Nodal (collateral) sprouting was not changed.