The upregulation of acetylcholine release at endplates of alpha-bungarotoxin-treated rats: its dependency on calcium.

1. The presynaptic component of an adaptive feedback mechanism leading to increased acetylcholine (ACh) release was studied in endplates of diaphragms from rats treated chronically with alpha-bungarotoxin (alpha BTX). 2. Quantal contents were calculated 'directly' from the amplitude of miniature endplate potentials (MEPPs) and endplate potentials (EPPs) which were recorded after mu-conotoxin treatment to prevent muscle action potentials. 3. In vitro application of the Ca2+ channel blockers nifedipine (10 microM) or omega-conotoxin (40 nM) had no significant effect on the increased quantal content of endplates from alpha BTX-treated rats. 4. At control endplates, in vitro block of presynaptic K+ channels by 5 microM 3,4-diaminopyridine did increase the quantal content to a level which was similar to that found in endplates of alpha BTX-treated rats but also induced a broadening of EPPs, which was not found at endplates after alpha BTX treatment. 5. The difference between quantal contents of alpha BTX-treated and control rats was highly dependent on the [Ca2+]o/[Mg2+]o ratio when [Mg2+]o was fixed at 1 mM. At low [Ca2+]o, the quantal content of endplates from alpha BTX-treated rats was lower than that of controls while at [Ca2+]o in the normal and high range this was reversed. However, changing the [Ca2+]o/[Mg2+]o ratio by means of [Mg2+]o, at a fixed [Ca2+]o of 2 mM, did not influence the relative increase of quantal contents at endplates from alpha BTX-treated rats. Double logarithmic plots of the 'toxin-induced' myasthenia gravis (TIMG) and control quantal content versus [Ca2+]o had an approximately linear part between 0.2 and 1.5 mM [Ca2+]o. The slopes of the TIMG and control lines were 1.81 and 0.96, indicating that the ACh release in TIMG muscles was more sensitive to changes of [Ca2+]o than controls. 6. At normal [Ca2+]o and [Mg2+]o, the depression of EPP amplitude during stimulation of the phrenic nerve at 30-50 Hz was somewhat larger at endplates from alpha BTX-treated rats than at control endplates. At low [Ca2+]o, the potentiation of EPP amplitudes during a stimulus train was much larger at endplates from alpha BTX-treated rats than from controls. 7. The results do not support the idea that the increased release of ACh is caused via regulatory effects on the presynaptic Ca2+ or K+ channels. Instead, the anomalous dependency of ACh release on Ca2+ in muscles of alpha BTX-treated rats suggests that a cytoplasmic, Ca(2+)-dependent, component is involved in the adaptive change of transmitter release.