The distribution of cell bodies containing serotonin (5-HT)-, substance-P (SP)-, neurotensin (NT)-, and somatostatin (SS)-like immunoreactivity (IR) in ventrolateral medulla (VLM) of the cat was studied immunohistochemically after administration of colchicine into the cisterna magna. Perikarya containing 5-HT-, SP-, NT- or SS-IR were found throughout the rostrocaudal extent of the VLM. Although neurons containing the different neuroactive substances appeared to have an overlapping distribution in VLM, some distinct differences were observed. In the caudal VLM most of the immunoreactive cell bodies observed contained 5-HT-IR. These neurons were found primarily in the region medial to lateral reticular nucleus (LRN) around the exiting intramedullary rootlets of the hypoglossal nerve (12N). In the intermediate region of VLM, perikarya containing 5-HT- and SP-IR were observed primarily near the ventrolateral surface of the medulla in the region around the exiting rootlets of the 12N. In contrast, most of the cells containing NT- and SS-IR were consistently observed to occupy a region in the medullary reticular formation immediately dorsal to that where 5-HT- and SP-IR perikarya were found. Finally, most of the immunoreactive perikarya were found in the rostral VLM; perikarya containing 5-HT- and SP-IR were observed throughout the nucleus paragigantocellularis lateralis (PGL) near the ventrolateral surface of the medulla. These data indicate that neurons immunoreactive to either 5-HT or several different neuropeptides were located in regions of VLM which have previously been implicated in the control of arterial pressure. As regions of VLM containing these neuroactive substances in neuronal perikarya have been shown to have direct connections with spinal sympathetic areas it is likely that these VLM cells are components of neuronal circuits involved in homeostatic mechanisms controlling the circulation.