Microinjections of the excitatory amino acid L-glutamate were made into the rostral ventrolateral medulla (RVLM) of anesthetised cats, to map the sites at which selective stimulation of cell bodies elicited a significant antinociceptive response (> or = 15% inhibition of the increase in L7 ventral root activity reflexly evoked by stimulation of C-fiber afferents). Antinociceptive sites were largely confined to the RVLM subregion ventromedial to the retrofacial nucleus, extending from the caudal pole of the facial nucleus to the level approximately 2.5 mm more caudal. Increases in arterial pressure were also elicited from some sites in the RVLM, but these were mainly lateral to the antinociceptive sites. In a second series of experiments, rhodamine labeled microspheres or cholera toxin B-gold (CTB-gold) were injected into the dorsal horn of the L7 segment. In three of these experiments in which the injection sites were restricted to the dorsal horn, retrogradely labeled cells in the caudal pons and medulla were virtually all within either the nucleus raphe magnus or the RVLM. Furthermore, the labeled cells in the RVLM were virtually confined to a discrete group located just ventromedial to the retrofacial nucleus, i.e. within the antinociceptive region as mapped by glutamate microinjection. The results of the present study indicate that antinociceptive effects are elicited by stimulation of a subregion in the RVLM, which is located medial to the pressor region. Further, the antinociceptive effects may be mediated, at least in part, by cells projecting directly to the dorsal horn in the spinal cord.