The major purpose of the present study was to determine the origin of the serotonin-containing neurons which project to the rostral ventral respiratory group in the rat. This was accomplished by using the technique of retrograde tracing with rhodamine-labeled latex microspheres (beads) combined with immunochemistry. The rhodamine-labeled beads were microinjected into electrophysiologically identified groups of inspiratory neurons in the rostral ventral respiratory group to retrogradely label neurons projecting to this site. Immunohistochemical processing of the tissue was then done to determine if serotonin was present in the retrogradely-labeled neurons. Serotonin-containing neurons projecting to the rostral ventral respiratory group were found in the raphe magnus, raphe obscurus, raphe pallidus and in the paraolivary region extending to the ventral medullary surface. No serotonin-containing neurons in more rostrally located raphe nuclei were found to project to the rostral ventral respiratory group. The findings suggest that caudal raphe serotonergic projections may affect the activity of respiratory neurons in the rostral ventral respiratory group. Projections to the rostral ventral respiratory group from other pontomedullary nuclei were also identified. Rhodamine-labeled neurons were found in the area of the Kölliker-Fuse nucleus, lateral and medial parabrachial nuclei, retrofacial nucleus, nucleus ambiguus/retroambigualis, nucleus tractus solitarius, A5 region, nucleus paragigantocellularis lateralis, retrotrapezoid nucleus, area postrema and spinal trigeminal nucleus. The projections to the rostral ventral respiratory group in the rat are similar to those previously described in the cat and suggest a common circuitry for the CNS control of breathing.