Three prepro-gonadotropin-releasing hormones, seabream GnRH (sbGnRH), chicken GnRH-II (cGnRH-II), and salmon GnRH (sGnRH) were isolated by cDNA cloning from the brain of the Atlantic croaker, Micropogonias undulatus. The amino acid sequences of croaker GnRH precursors show greatest similarities to those of the gilthead and red sea breams and European sea bass. In situ hybridization of croaker brain sections revealed more abundant sbGnRH mRNA expression in the preoptic area (POA) than in other brain regions. sbGnRH mRNA expression was also observed in the olfactory bulb (OB; but not in the terminal nerve ganglion cells [TNgc]), ventral telencephalon (vTEL), and anterior hypothalamus. In addition, specific sbGnRH mRNA signals were detected in the pituitary. cGnRH-II mRNA expression was limited to the midbrain tegmentum. Neuronal elements expressing sGnRH mRNA were detected in the OB including the TNgc, vTEL, and POA, indicating an overlap of the sbGnRH and sGnRH systems in certain ventral forebrain areas. The results of quantitative reverse transcriptase-polymerase chain reaction of the three GnRH mRNAs in different brain areas and the pituitary are consistent with their localization by in situ hybridization. Interestingly, a few sbGnRH mRNA-expressing neuronal elements were observed arranged in a row in the anteroventral hypothalamus projecting toward the pituitary. The results provide a morphological basis for a putative role of sbGnRH as the gonadotropin-releasing hormone. Moreover, localization of sbGnRH mRNA in a teleost pituitary points to sbGnRH synthesis, and its potential role as a local regulator, within the pituitary, similar to the role of GnRH-I in mammals.