The tryptophan hydroxylase inhibitor para-chlorophenylalanine (pCPA) was employed in an investigation of the role of brain serotonin in the regulation of pituitary cyclic AMP levels in Salmo gairdneri. Hypothalamic serotonin content was measured with a fluorometric assay, while pituitary cAMP was determined by means of a double-antibody radioimmunoassay. A time-course study revealed that 250 mg pCPA/kg significantly reduced hypothalamic serotonin content within 1 hr after injection. Lower doses were effective in reducing serotonin content after 48 hr. Pituitary cAMP levels declined significantly 1 hr after injection of 250 mg pCPA/kg. Administration of pCPA at 50 and 250 mg/kg doses reduced cAMP levels within 24 hr. These data indicate that a serotonergic mechanism is involved with the regulation of pituitary cyclic AMP levels in the rainbow trout.