Abstract: The effects of several phospholipase A~2~ neurotoxins from snake venoms were examined on purely cholinergic synaptosomes from Torpedo electric organ. The noncatalytic component A of crotoxin had no effect, whereas its phospholipase component B, used alone or complexed to component A, elicited a rapid and dose‐dependent acetylcholine (ACh) release and a depolarization of the preparation. Subsequent ACh release evoked by high K^+^ levels or calcium ionophore was identical to the control after the action of component A but reduced after the action of crotoxin or of component B. These effects were not observed when the phospholipase A~2~ activity of the toxin was blocked either by replacing Ca^2+^ by Ba^2+^ (respectively, activator and inhibitor of phospholipase A~2~) or by alkylation of component B with p‐bromophenacyl bromide. β‐Bungarotoxin, another very potent phospholipase A~2~ neurotoxin, induced release of little ACh, did not affect ionophore‐evoked ACh release, but significantly reduced depolarization‐induced ACh release. The single‐chain phospholipase A~2~ neurotoxin agkistrodotoxin behaved like crotoxin component B. A nonneurotoxic phospholipase A~2~ from mammalian pancreas induced release of an amount of ACh similar to that released by crotoxin but did not affect the evoked responses. The obvious differences in effect of the various neurotoxins suggest that they exert their specific actions on the excitation‐secretion coupling process at different sites or by different mechanisms.