The aminosteroid U73122 inhibited phospholipase C (PLC)-mediated intracellular Ca2+ release in differentiated and undifferentiated NG108-15 cells, as well as rat dorsal root ganglion (DRG) neurons grown in primary culture. 1 microM U73122 blocked bradykinin (BK)-induced increases in the intracellular free Ca2+ concentration ([Ca2+]i) measured in single cells with indo-1-based dual emission microfluorimetry. A close structural analog, U73343, was without effect. The effects of U73122 were time and concentration-dependent. 1 microM drug produced half maximal inhibition in approximately 3 min. The IC50 for a 20-min exposure was approximately 200 nM. The effects of the compound were irreversible for the duration of experiments as long as 1 h. Treatment with 1 microM U73122, but not U73343 produced a small but significant increase in [Ca2+]i which resulted from Ca2+ release from an intracellular store. It is not clear whether this [Ca2+]i increase resulted from inhibition of PLC or an action on the store directly. In differentiated NG108-15 cells U73122 blocked completely depolarization-induced Ca2+ influx. In contrast, in DRG neurons U73122 inhibited only slightly voltage-sensitive Ca2+ channels. Thus, we caution that U73122 may not be selective at concentrations required for maximal block of PLC and that the selectivity of U73122 is dependent on cell type. Overall, our results are consistent with U73122 inhibiting PLC in neuronal cells and indicate that under the appropriate conditions, this compound is a useful tool for studying inositol 1,4,5-trisphosphate (IP3)-mediated Ca2+ mobilization.