When acetylcholinesterase was inhibited by neostigmine, SK&F 96365 (1-(beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl)-1H-imidazole hydrochloride) at 10 microM caused no effect on the amplitude of single endplate potentials (e.p.p.s) but shortened the decay time in mouse phrenic nerve-diaphragm preparations. However, SK&F 96365 inhibited high-frequency stimulation-evoked long-lasting depolarization of the endplate region and accelerated the run-down of trains of e.p.p.s which were eliminated within 1 s. After a train of stimulation, SK&F 96365 produced a post-tetanic depression of single e.p.p.s. The post-tetanic effect gradually dissipated with full restoration in 10-15 s. During a train of stimulation, SK&F 96365 also depressed miniature endplate potentials (m.e.p.p.s), which were restored after termination of stimuli in parallel with the recovery of e.p.p. The decay times of miniature endplate currents during recovery phases changed slightly. In control preparations not treated with neostigmine, however, SK&F 96365 did not alter the amplitude and decay time of m.e.p.p.s or e.p.p.s but accelerated the decay of succinylcholine-induced endplate depolarizations. The results suggest that SK&F 96365 facilitates nicotinic receptor desensitization in addition to blocking receptor-operated Ca2+ channels.