In situ hybridization histochemistry was used to map the distribution of alpha 2, alpha 3, alpha 4, and beta 2 nAChR subunit mRNAs throughout the peripheral vestibular system of the rat. The alpha 4 and beta 2 nAChR subunit genes were co-expressed by populations of primary afferent neurons within Scarpa's ganglion, while there was no expression of the alpha 2, alpha 3, alpha 4, or beta 2 nAChR subunit genes by type I or type II vestibular hair cells. alpha-bungarotoxin binding to nAChRs in the vestibular end-organs was primarily limited to the afferent chalices surrounding type I hair cells and the basal aspect of type II hair cells. These data suggest that nAChRs composed of alpha 4 and beta 2 subunits are localized on afferent chalices innervating the type I vestibular hair cells and that the direct cholinergic efferent innervation of the type II vestibular hair cells utilizes nAChR composed of other subunits.