Acetylcholine induces a complex electrical membrane response in Xenopus laevis oocytes. This response is mimicked, and probably mediated by injected inositol 1,4,5-trisphosphate. Oocytes prelabelled with 45Ca released calcium in two phases, the second, slow phase exhibiting first order kinetics of release. Brief exposure of prelabelled oocytes to acetylcholine resulted in a significant increase in the rate of calcium release that returned to control values 2-3 min following the removal of the neurotransmitter. Intracellular injection of inositol 1,4,5-trisphosphate resulted in increased rate of calcium release similar to, but longer than that caused by acetylcholine. Experiments conducted on single oocytes permitted the investigation of the relationship between acetylcholine-induced and inositol 1,4,5-trisphosphate-induced calcium mobilization and the resulting electrical membrane response. Our data reinforce our previous suggestion that inositol 1,4,5-trisphosphate is the intracellular second messenger of the muscarinic membrane electrical response in Xenopus oocytes.