Retrograde axonal transport of fluorescent tracers (primuline, FluoroGold and Nuclear Yellow) from the spinal cord, frontal cortex, lateral hypothalamus to various neuronal groups of the midbrain periventricular gray substance (periaqueductal gray matter) and to the dorsolateral pontine tegmentum in the rat has been studied. Two large groups of serotonin-containing neurons have been found to be localized in the dorsomedial region of the nucleus raphe dorsalis. They are sources of projections into the thoracic segments of the spinal cord. A part of these neurons gives divergent axon collaterals to the frontal cortex and to the spinal cord. Non-collateral projections of the dorsolateral pontine tegmental catecholaminergic neurons to the spinal cord and the frontal cortex have been revealed. The data obtained give support to the fact that antinociceptive effect of stimulation of the "pure analgesic zone" [Fardin et al. (1984) Brain Res. 306, 105-123.] of the midbrain periaqueductal gray matter may be due to direct involvement of the nucleus raphe dorsalis into a descending control over transmitting nociceptive stimuli at the spinal cord level. Neurotransmissive and neuroregulatory roles of separate cortical, hypothalamic and spinal serotonergic projections of the nucleus raphe dorsalis neurons are discussed.