Serotonin-synthesizing raphé/parapyramidal neurons (5-HT neurons) may function as sympathetic premotor neurons regulating sympathetic outflow to the cutaneous vascular bed. In the present study a genetically engineered pseudorabies virus (PRV) expressing green fluorescent protein (GFP) was injected into the rat tail. After survival for 3–4 days the medulla oblongata was examined using double-label immunohistochemistry, with an antibody against GFP for the virus and an antibody against phenylalanine hydroxylase 8 (PH8) for 5-HT synthesis. Sections were examined using light microscopy, and conventional and confocal fluorescence microscopy. There were two subpopulations of PRV+ve neurons in the raphé/parapyramidal region: a more dorsally and laterally located subgroup of medium-sized and large neurons, mainly non-serotonergic, and a more ventrally located subgroup of small mainly serotonin-synthesizing neurons, including those just dorsal to the pyramids, those in raphé pallidus, and those in close relationship to the ventral surface in the parapyramidal-subependymal zone.