Calcium regulation is an important event in synaptic transmission and neuronal function, which is governed by a very intricate signal transduction system which is not completely understood. Using a variety of pharmacological assays, we have characterized the action of deltamethrin on the ciliary voltage-sensitive calcium channel and on phospholipase C activity of Paramecium tetraurelia Sonneborn, an organism that does not possess a voltage-sensitive sodium channel. In fura-2 fluorometric assays, which examined whole cells and ciliary membrane vesicles enriched with calcium channels, deltamethrin stimulated Ca2+ uptake. We also determined that the phospholipase C activity of the ciliary membrane vesicles is regulated by the βγ-subunit from heterotrimeric G-proteins. Subsequent treatment with deltamethrin resulted in a substantial and highly significant increase in phospholipase C activity. These results provide evidence that the molecular mode of action of pyrethroids on the voltage-sensitive calcium channel is distinct from the action of this insecticide on the voltage-sensitive sodium channel and may be dependent, in part, upon an interaction with the βγ-subunit of heterotrimeric G-protein.