The purpose of the present work was to characterise the effects of trimethyltin on the release of acetylcholine from parasympathetic nerves and its effect on the postjunctional cholinergic stimulation of a smooth muscle. The guinea-pig trachea has been used as a model. Prejunctionally, trimethyltin (3.0 × 10(-3) M) significantly enhanced in a reversible manner the high K(+) (75 mM) evoked release of endogenous acetylcholine and [(3)H]acetylcholine. The evoked release of endogenous acetylcholine and [(3)H]acetylcholine was released from a pool of acetylcholine being independent of extraneuronal Ca(2+) in the presence, but not in the absence of trimethyltin. The effect of trimethyltin on the release was not inhibited by low Ca(2+) (0 mM and 1.0 × 10(-4) M) or by Ca(2+) channel blockers (verapamil, 1.0 × 10(-4) M, flunarizine, 1.0 × 10(-4) M, ω-conotoxin GVIA, 2.0 × 10(-7) M and ω-agatoxin, 2.0 × 10(-7) M). The present results also demonstrate that trimethyltin induce emptying of a non-vesicular, probably a cytoplasmic storage pool of acetylcholine, since AH5183 (2.0 × 10(-5) M), an inhibitor of the translocation of acetylcholine into synaptic vesicles, and α-latrotoxin (1.0 × 10(-8) M), a toxin from black widow spider venom inducing vesicle depletion, had no inhibitory effects on the release of [(3)H]acetylcholine evoked by trimethyltin (3.0 × 10(-3) M). The release of [(3)H]acetylcholine was moreover enhanced by trimethyltin when the vesicular uptake of [(3)H]acetylcholine was inhibited by AH5183, probably as a result of a higher cytoplasmic concentration of [(3)H]acetylcholine. Trimethyltin also reduced the neuronal uptake of [(3)H]choline and this was probably due to a depolarising effect of trimethyltin on the cholinergic nerve terminals. A similar depolarisation induced by trimethyltin was observed during patch clamping of GH(4) C(1) neuronal cells. Postjunctionally, trimethyltin had no effect by itself or on the carbachol-induced smooth muscle contraction, indicating that trimethyltin did not have a general depolarising effect on smooth muscle cells or an effect on muscarinic receptors. Furthermore, the reduced electrical field-induced contraction and the subsequent increase in the basal smooth muscle tension that was observed by addition of trimethyltin was activity-dependent, and was most probably due to emptying of a nervous non-vesicular storage pool of acetylcholine, followed by rapid hydrolysis of acetylcholine by acetyl- and pseudocholinesterases.