Pheromones affect gonadal functions and sexual behaviors. Information in regard to pheromones is received by the vomeronasal organ (VNO) and transmitted to the accessory olfactory bulb (AOB). We investigated the physiological role of the alpha1B and beta3 subunits of the N (neuronal)-type voltage-dependent Ca2+ channel in the neurotransduction in the accessory olfactory (vomeronasal) system using alpha1B-deficient mice and beta3-deficient mice. RT-PCR studies showed the existence of beta1, beta2, beta3, beta4, alpha1A, alpha1B, and alpha1C subunits of voltage-dependent Ca2+ channels in the mouse VNO. Immunohistochemical studies showed that the alpha1A, alpha1B, and alpha1C subunits of voltage-dependent Ca2+ channels exist in the sensory neurons and supporting cells of the mouse VNO. Exposure of the VNO to urine samples excreted from male mice induced lower Fos-immunoreactivity in the periglomerular (PG) cells of the AOBs in alpha1B-deficient female mice than in those of wild mice. The density of Fos-immunoreactive (Fos-ir) cells after exposure to female urine samples at the periglomerular cell layer of alpha1B-deficient male mice was lower than that of wild mice. Exposure of the VNO of beta3-deficient female mice to male urine samples also induced low Fos-ir cells in the periglomerular cell layer of the AOB. These data suggest the importance of the alpha1B and beta3 subunits of the N-type voltage-dependent Ca2+ channel for the pheromone signal transduction system.