Immunostaining of brain and pituitary sections of three teleost species (goldfish, trout and eel) with antisera to porcine and human ACTH 1-39 revealed the presence of an ACTH (adrenocorticotropic hormone)-like peptide in the ventral hypothalamus. Perikarya were localized in the rostral, median and posterior portions of the nucleus lateralis tuberis (NLT); some were in contact with the cerebrospinal fluid. A dense network of immunoreactive (ir) fibers occurred in the peri-infundibular region and extended into the periventicular tissue, around the lateral and posterior recesses. Rostrally directed ir-fibers reached the telencephalon either ventrally or mediodorsally; some were observed in the olfactory lobe. In the mesencephalon, ir-fibers penetrated into the optic tectum of the goldfish. In the pituitary, both antisera intensely labeled rostral ACTH cells. Small groups of labeled cells were scattered in the rostral pars distalis and the proximal pars distalis. A gradient of activity was evident among ACTH cells: those located along the rostral neurohypophysis containing corticotropin-releasing factor nerve terminals were larger and often more marked than those farther away from the neural tissue. ACTH-like peptide in the brain may act as a neuromodulator, mainly in the NLT and the preoptic nucleus, and around the nuclei of the ventricular recesses containing serotonin and catecholamines.