Three forms of gonadotropin-releasing hormone in a perciform fish (Sparus aurata): Complementary deoxyribonucleic acid characterization and brain localization

Three forms of GnRH-salmon (sGnRH), seabream (sbGnRH), and chicken (cGnRH-II)-have been described in the gilthead seabream (Sparus aurata) brain, and the cDNA encoding the sbGnRH precursor was recently isolated. In the present study, the cDNAs encoding the sGnRH and cGnRH-II were isolated and characterized, and the neurons producing the three GnRHs were localized in the seabream brain. Fragments of sGnRH and cGnRH-II cDNAs were amplified by polymerase chain reaction and used as probes to isolate the full-length cDNAs from a brain cDNA library. The cDNA encoding the cGnRH-II precursor is 573 nucleotides (nt) long, and the cDNA encoding the sGnRH precursor is 1971 nt in length with an unusually long 5' untranslated region. Specific single-strand DNA probes for in situ detection of mRNA were designed according to nonconserved regions among the three GnRH cDNAs. Localization of GnRH mRNA-producing cells in the brain revealed five distinct populations of cells: sGnRH-producing cells in the ventromedial olfactory bulbs and the terminal nerve, sbGnRH-producing cells in the preoptic area and the ventral thalamus, and cGnRH-II-producing cells in the midbrain tegmentum. The discrete sites of expression of the three forms of GnRH indicate that only sbGnRH is directly involved in the control of gonadotropin secretion.