The olfactory organ is a site for neuroendocrine modulation of reproduction in zebrafish

Gonadotropin-releasing hormone (GnRH) is one of the most fascinating neuroendocrine peptides: It is essential for maintaining the reproductive state of vertebrates and also displays high sequence homology to the tridecapeptide mating pheromone of the yeast, S. cerevisiae. In spite of its highly conserved role in vertebrate reproduction, recent studies in zebrafish show that the loss of function of genes encoding Gnrh isoforms does not cause infertility. Here we first investigated whether Phoenixin, a novel peptide acting in the reproductive pathway of vertebrates, is the hormone that has replaced Gnrh in zebrafish. While loss of function of the phoenixin gene affected female differentiation, we observed no defects in fertility. We next reconsidered the GnRH pathway and turned to the natural world, where fishes use waterborne hormones to control reproduction. Thus, we investigated whether exogenous Gnrh affects the hypothalamic-pituitary axis. Here we show that fish isolated from their conspecifics and kept in artificial water devoid of fish odors, mounted sex-appropriate pituitary responses when Gnrh was added to the water. Furthermore, blocking the access of water to the olfactory organs eliminated these responses. We then analyzed Gnrh signaling by knocking out the gene encoding gnrh-receptor3 (gnrh-r3) and, surprisingly, found that fish homozygous for a gnrh-r3 null mutation were almost completely infertile: males did not produce sperm and females produced but a few mature oocytes. Finally, we found that Gnrh was present in nanomolar concentrations in the water that houses the fish, thus supporting the hypothesis that waterborne Gnrh from conspecifics plays a key role in regulating zebrafish reproduction in the absence of the endogenous ligand.