Concentrations of pituitary adenylate cyclase activating polypeptide (PACAP) in 79 microdissected human brain regions from adult 40- to 80-year-old adult males were measured by radioimmunoassay. Although PACAP was detectable in all of the brain nuclei investigated, the distribution of the peptide was regionally very heterogeneous. The ratio of the highest to the lowest concentrations measured in brain regions exceeded 160:1. The highest concentrations were found in the dorsal vagal complex, the bed nucleus of the stria terminalis, the median eminence-pituitary stalk, and in the periventricular and paraventricular hypothalamic nuclei. They were followed by some hypothalamic (supraoptic and ventromedial), preoptic and brainstem nuclei. High concentrations were also measured in the septum pellucidum, periaqueductal and spinal gray matters, the motor facial, and in the spinal nucleus of the trigeminal nerve. The distribution pattern of PACAP in the human brain was unique; it did not show any similarities to the distributions of other neuropeptides in the central nervous system.