Adrenergic neurons in the C1 cell group in the rostral ventrolateral medulla oblongata contain epinephrine, as well as its biosynthetic enzyme, phenylethanolamine N-methyltransferase (PNMT). These neurons send axons to regions of the central nervous system known to regulate autonomic function, including the sympathetic preganglionic nuclei of thoracic and upper lumbar spinal cord. Previous studies have shown that PNMT is expressed in neurons located in the medulla oblongata on embryonic day 14; however, the development of the projections from these cells has not been studied. With the aid of high-performance liquid chromatography (HPLC) to determine levels of catecholamines and immunocytochemistry to demonstrate PNMT, the ontogeny of the adrenergic bulbospinal pathway in the embryonic, postnatal, and adult rat has been studied. In addition, the relationship between PNMT-immunoreactive (IR) fibers and retrogradely labeled sympathetic preganglionic neurons projecting to adrenal medulla are described. PNMT-IR fibers were first observed in the caudal medulla oblongata and lateral funiculus of spinal cord on gestational day 15(E15). On E16, PNMT-IR fibers in the thoracic spinal cord were observed in the intermediate gray matter at the level of the lateral horn. Epinephrine was measureable in spinal cord on E20. Both the density of PNMT-IR fibers and the levels of epinephrine increased to a maximum during the second postnatal week and then declined to adult levels. These observations suggest that a period of adrenergic hyperinnervation of spinal sympathetic nuclei occurs during the neonatal period. PNMT-IR terminals in spinal cord were observed, primarily, although not exclusively, in sympathetic nuclei of thoracic cord and parasympathetic nuclei of upper sacral cord. Adrenergic fibers in the intermediolateral nucleus (IML) and the central autonomic nucleus (CAN) dorsal to the central canal were particularly dense during the second postnatal week in both midthoracic and upper sacral segments. In the neonate, a "ladder-like" pattern of PNMT-IR fiber staining was observed which represented transverse fiber bundles connecting IML with CAN and extensive longitundinal fiber bundles along the border of the funiculus in IML. At all spinal levels, adrenergic fibers were also observed adjacent to the ependyma dorsal or lateral to the central canal. The relationship between adrenal preganglionic neurons and PNMT-IR fibers in IML was examined on postnatal days 4, 15, and 60. With retrograde labeling from adrenal medulla, it was demonstrated that PNMT-IR fibers are associated with adrenal preganglionic neurons throughout postnatal development.(ABSTRACT TRUNCATED AT 400 WORDS)