Dopamine-beta-hydroxylase (DBH), the enzyme that converts dopamine to norepinephrine, has been localized in light and electron microscopic preparations of rat brain by an immunocytochemical method using a peroxidase--anti-peroxidase Fab complex. In light microscopic preparations, DBH-specific reaction product was observed in somata and proximal processes of neurons in the locus coeruleus and subcoeruleus as well as within distal axons of the principal adrenergic fiber system. DBH-specific reaction product was also observed within small (1--2 micrometer), punctate structures in the interstitial nucleus of the stria terminalis and the para- and periventricular nuclei of the hypothalamus. Electron microscopic results demonstrated on association of DBH-specific reaction product with the Golgi apparatus of neuronal somata in the locus coeruleus and subcoeruleus. DBH-positive reaction product was also seen in association with small (35-55 nm) agranular synaptic vesicles and large (80--100 nm), probable granular vesicles within axonal varicosities and terminals in the interstitial nucleus of the stria terminalis. Occasionally, DBH-containing axonal varicosities and terminals were observed to form synapse-like junctions with dendritic profiles, but most of the observed DBH-positive axonal structures did not establish identifiable synaptic relationships.