In NG 108-15 clonal cells, extracellular application of micromolar concentrations of serotonin [5-hydroxytryptamine (5-HT)] and substance P induces the opening of a cation permeability monitored by the influx of [14C]-guanidinium. The serotoninergic component of this cation permeability is linked to 5-HT3 receptor activation, whereas the substance P component probably involves an "N-terminal-dependent substance P receptor." In this study, [14C]guanidinium influx triggered by 1 microM 5-HT plus 10 microM substance P was shown to be insensitive to tetrodotoxin, verapamil, diltiazem, nimodipine, and omega-conotoxin, as expected from a process independent of voltage-sensitive sodium and calcium channels. In contrast, [14C]guanidinium influx was inhibited by millimolar concentrations of extracellular calcium and by the chelation of intracellular calcium by bis-O-aminophenoxyethanetetraacetic acid. The inhibition by extracellular calcium apparently involved a competition between the divalent cation and [14C]guanidinium for the same channel. When NG 108-15 cells were exposed to X537A, an ionophore that specifically induces release of calcium from intracellular stores, [14C]guanidinium uptake was markedly increased even in the absence of 5-HT and/or substance P. Conversely, [14C]guanidinium influx due to the latter substances could be reversibly and dose-dependently blocked by various drugs that possess calmodulin-antagonizing properties. These results strongly suggest that the cation permeability opened by 5-HT and substance P in NG 108-15 cells involves a calcium/calmodulin-dependent process.(ABSTRACT TRUNCATED AT 250 WORDS)