Dual excitation microfluorimetry (Fura-2) was used to measure changes in intracellular calcium ([Ca2+]i) in individual cultured guinea pig myenteric neurons. Bombesin (5-500 nM) induced concentration-dependent increases in [Ca2+]i responses, with a maximal effect at 500 nM (56% of neurons responding, mean peak Ca2+ response 244 +/- 25 nM vs. basal 65 +/- 7 nM). Removal of Ca2+ from the median did not affect the initial [Ca2+]i peak but eliminated the subsequent plateau phase. The [Ca2+]i responses to bombesin was abolished by preincubation with thapsigargin (1 microM), a Ca(2+)-ATPase inhibitor (91 +/- 7% inhibition). [Ca2+]i responses to bombesin were inhibited by U73122 (1 microM), an inhibitor of phospholipase C (84 +/- 6% inhibition).