Calcium dependence of quantal release triggered by graded depolarization pulses to nerve terminals on crayfish and frog muscle.

Quantal transmitter release was measured in small portions of neuromuscular junctions by means of a perfused macro-patch-clamp electrode. Release was elicited by graded current pulses through the recording electrode (excitation blocked by TTX). On increasing the stimulation current from a threshold amplitude, release rose steeply for several orders of magnitude and finally approached a saturation level of about 10 quanta/pulse. Reduction of the Ca concentration in the perfusate of the electrode, Cae, depressed the saturation level of release relatively little and had practically no effect on the threshold current amplitude, as long as the Ca concentration in the superfusion of the bath, Cab, remained high. When Cab was reduced too, the depression of release was more severe. The dependence of release on Cae was determined for a large range of Cae for saturating depolarization pulses. In crayfish, at 0 Cab, in double-logarithmic release-Cae plots the maximum slope was on average 3.9, and this slope dropped to on average 2.1 in 13.5 mM Cab. In frog, at 0 Cab, the respective double-logarithmic slope was 3.5, while in 1.8 mM Cab this slope declined dramatically, the rate of release decreasing on average only by a factor of 3.8 from 10 mM to 0.02 mM Cae. These results are interpreted by the assumption that the resting Ca concentration in the terminal, Cair, has strong influence on the rate of release due to depolarization pulses in low Cae, and that Cab has control on Cair in the terminal.