The distribution of Pituitary adenylate cyclase-activating polypeptide (PACAP) was investigated in the brain, pituitary and sensory organs of the zebrafish, Danio rerio, during development, in juvenile and adult specimens, using the immunofluorescence method. In 24 h post fertilization (hpf) embryos, PACAP immunoreactive cells appeared in the rostral telencephalon, dorsal diencephalon, caudal and medial rhombencephalon, spinal cord and retina. At 48 hpf stage, positive cells were present in the dorsal diencephalon, medial rhombencephalon, spinal cord, retina and olfactory placode (Op). At 72 hpf stage, additional immunoreactive elements appeared in the medial telencephalon, hypothalamus, mesencephalic tegmentum, retina and otic sensory epithelium (Ose). At day 5, new immunoreactive cells were found in the anterior rhombencephalon and pituitary pars distalis. At day 13, positive cells were mainly concentrated in the mesencephalic tegmentum and spinal cord. In the telencephalon, diencephalon, rhombencephalon and pituitary, the distribution of positive cells was similar to that previously reported. At 1 month stage, positive cells were detected in the hypothalamus, nucleus of the medial longitudinal fascicle (nMlf), rhombencephalic griseum centrale (Gc) and pituitary pars distalis. At 2-3 month stages, immunoreactive elements were found in several hypothalamic nuclei, in the mesencephalic nucleus isthmi, cerebellum and pituitary. In adults, PACAP immunoreactivity was confined to a few brain regions and the pituitary. PACAP immunoreactivity was transiently expressed in several regions suggesting that the peptide may have a role in the control of cells differentiation and proliferation during zebrafish ontogeny. The finding of positive fibers in the pituitary from day 5 onward indicates that PACAP may function from this stage as a hypophysiotropic peptide.