The distribution of immunoreactive (ir) atrial natriuretic peptides (ANPs) in 47 microdissected brain and spinal cord regions of the rat was determined by radioimmunoassay. The highest concentrations of ir-ANPs exist in the paraventricular nucleus and median preoptic nucleus (580.9 and 558.0 fmol/mg protein, respectively). High concentrations of ir-ANP (greater than 300 fmol/mg protein) are present in the interpeduncular nucleus, preoptic and hypothalamic periventricular nuclei, median eminence and organum vasculosum of the lamina terminalis. Moderate concentrations of ir-ANPs (between 100 and 300 fmol/mg protein) are found in 16 brain regions such as the bed nucleus of the stria terminalis, nucleus of the diagonal band, most of the hypothalamic nuclei, central gray, locus coeruleus and parabrachial nuclei. Low levels of ir-ANPs (less than 100 fmol/mg protein) exist in 22 brain regions including cortical areas, amygdala, caudate nucleus, nucleus accumbens, hippocampus, supraoptic nucleus, subfornical organ, medial mammillary nucleus, substantia nigra, dorsal raphe nucleus, cerebellum, nucleus of the solitary tract and others. Cervical spinal cord and neurointermediate lobe of pituitary gland contain low levels of ir-ANPs.