The ultrastructural distribution of calcitonin gene-related peptide immunoreactivity (CCRPi) was examined in sections of decalcified temporal bones in order to study the complex peptidergic innervation patterns of this efferent neuromodulator in the peripheral vestibular system of the rat. A new method of preembedding immunoelectron microscopy was developed to accomplish this study. Unmyelinated CGRPi axons, measuring 1 to 3 jam in diameter, passed among the primary afferent fibers in Scarpa's ganglion, and these fibers continued through the subepithelial regions of the vestibular end-organs. Within the neurosensory epithelia of the maculae and cristae, the CGRPi axons ramified to produce numerous CGRPi terminals. Immunoelectron microscopic localization of CGRPi terminals in the maculae and cristae revealed an extensive innervation pattern on the afferent vestibular pathway. Calcitonin gene-related peptide immunoreactive terminals made synaptic contacts with the unmyelinated portions of the primary afferent vestibular fibers innervating both type I and type II hair cells. Abundant synaptic contact between CGRPi terminals and the chalices surrounding type I hair cells was observed. Rare direct contact between CGRPi terminals and type I or type II hair cells was observed. In addition, vesiculated efferent terminals without CGRPi were seen contacting type II hair cells. These data suggest that the efferent vestibular system has a much more complex innervation pattern on the afferent vestibular pathway than previously believed