The calcium dependence of spontaneous and evoked quantal release at the frog neuromuscular junction

Abstract

1. The quantal output from stimulated nerve terminals in the frog sciatic nerve-sartorius muscle preparation in low-Ca(2+) Ringer solution was measured by the coefficient of variation and the failures methods. Adding sucrose to the Ringer to increase the tonicity or adding ethanol increased miniature end-plate potential (m.e.p.p.) frequency and also the end-plate potential (e.p.p.) amplitude. Earlier reports suggested that increases in tonicity did not increase evoked quantal release.2. Concanavalin A has been reported to block the increase in m.e.p.p. frequency caused by increasing the tonicity of the Ringer (Gorio & Mauro, 1979). This effect was confirmed. The lectin-treated preparations also failed to show an increase in evoked quantal release when the tonicity was increased.3. A model in which both spontaneous and evoked quantal releases depend on some power of the intracellular [Ca(2+)] is presented. The model predicts that rises in m.e.p.p. frequency will be accompanied by increased quantal output from stimulated nerve terminals. The maximum slope of the relationship between log (evoked quantal output) and log ([Ca(2+)](out)) will be less than the true power. A theoretical analysis shows that, as the true power approaches infinity, the maximum slope will be slightly above 4. The value for the slope usually found experimentally at the frog neuromuscular junction is also about 4.4. The model does not fit the experimental data. The observed increases in evoked quantal release are higher than those predicted for the observed increases in spontaneous release. There are several possible explanations for the discrepancy. Treatments that increase m.e.p.p. frequency may also increase Ca(2+) influx into the stimulated terminal. However, we prefer the explanation that there is a fraction of spontaneous release that is independent of the [Ca(2+)] in the terminal; if this is true the model might account for the data.5. The model can account for a variety of puzzling experimental observations, including: (a) the effect of hypertonic solutions and of diamine in decreasing the slope in the relation between log (evoked quantal output) and log ([Ca(2+)](out)); (b) the slope of near 1 observed at the crustacean neuromuscular junction; (c) the decrease in the slope produced by treatment with botulinum toxin.