Mechanism of the calcium-dependent stimulation of transmitter release by 4-aminopyridine in synaptosomes.

The mechanism of the Ca2+-dependent stimulation of neurotransmitter release by 4-aminopyridine in synaptosomes was studied. The stimulation of gamma-[3H]aminobutyric acid and [3H]acetylcholine release by 4-aminopyridine was not significantly affected either by tetrodotoxin or by the absence of Na+ in the medium, whereas the toxin notably inhibited the release of both transmitters induced by veratridine. On the other hand, the release of labeled gamma-aminobutyric acid induced by 4-aminopyridine was inhibited by both La3+ and ruthenium red, two blockers of Ca2+ transport in synaptosomes. In other experiments, 4-aminopyridine had only a slight stimulatory effect, if any, on the influx of 45Ca2+ into synaptosomes, under both resting and K+-depolarizing conditions. Ruthenium red inhibited the stimulation by K+ of the 45Ca2+ uptake, and 4-aminopyridine did not antagonize this inhibition. We conclude that the transmitter-releasing action of 4-aminopyridine in synaptosomes does not involve an excitatory effect on the membrane which may result in the opening of voltage-sensitive Na+ channels. 4-Aminopyridine does not seem to act either by enhancing Ca2+ entry into the synaptosomes. It is proposed that 4-aminopyridine facilitates the coupling between Ca2+ binding and transmitter secretion at the presynaptic membrane.