Characterization and expression analysis of a GnRH-like peptide in the Pacific abalone, Haliotis discus hannai

Gonadotropin-releasing hormone (GnRH) is a key neuropeptide of vertebrates, involved in gonadal maturation and primarily regulated by the hypothalamic-pituitary-gonadal axis. The full length GnRH-like cDNA isolated from the cerebral ganglion of Haliotis discus hannai, encodes a deduced protein of 216 amino acids with a theoretical molecular mass of 23.36 kDa and an isoelectric point of 7.72. The GnRH transcript comprises a putative signal peptide, a GnRH dodecapeptide, a proteolytic processing site, and a GnRH associated peptide (GAP). Comparative structural analysis revealed that the cloned sequence shares relatively high homology with other molluscan species and a lower degree of amino acid identity with GnRH of piscine vertebrates. Phylogenetic comparison with other known GnRH genes revealed that the GnRH-like mRNA of H. discus hannai is most closely related to the marine gastropod, H. laevigata. Three-dimensional homology structure of H. discus hannai GnRH and GAP exhibited a helix-loop-helix structure. Quantitative PCR assay demonstrated wide expression of GnRH in all ganglia, among them cerebral ganglion exhibited the highest expression level. Significantly higher expression was observed in cerebral ganglion and gonadal tissues at higher effective accumulative temperature (1000 °C). In situ hybridization showed that the GnRH expressing neurosecretory cells distributed throughout the cortex of the cerebral ganglion. These results suggest that GnRH synthesized from the cerebral ganglia may be involved in gonadal maturation and regulating the secretion of other reproductive hormones.