Abstract
A full-length cDNA sequence encoding a GnRH receptor was cloned from the pleuropedal ganglion of the Pacific abalone, Haliotis discus hannai. The cloned sequence is 1499-bp in length encoding a protein of 460 amino acid residues, with a molecular mass of 52.22 kDa and an isoelectric point (pI) of 9.57. The architecture of HdhGnRH-R gene exhibited key features of G protein-coupled receptors (GPCRs), including seven membrane spanning domains, putative N-linked glycosylation motifs, and phosphorylation sites of serine and threonine residues. It shared 63%, 52%, and 30% sequence identities with Octopus vulgaris, Limulus polyphemus, and Mizuhopecten yessoensis GnRH-R II sequences, respectively. Phylogenetic analysis indicated that HdhGnRH-R gene was clustered with GnRH-R II of O. vulgaris and O. bimaculoides. qPCR assay demonstrated that the mRNA expression level of this receptor was significantly higher in the pleuropedal ganglion than that in any other examined tissue. Transcriptional activities of this gene in gonadal tissues were significantly higher in the ripening stage. The mRNA expression of this gene was significantly higher in pleuropedal ganglion, testis, and ovary at higher effective accumulative temperature (1000 °C). In situ hybridization revealed that HdhGnRH-R mRNA was expressed in neurosecretory cells of pleuropedal ganglion. Our results suggest that HdhGnRH-R gene synthesized in the neural ganglia might be involved in the control of gonadal maturation and gametogenesis of H. discus hannai. This is the first report of GnRH-R in H. discus hannai and the results may contribute to further studies of GPCRs evolution or may useful for the development of aquaculture method of this abalone species.
Highlights
Gonadotropin-releasing hormone receptors (GnRH-Rs) play crucial roles in mediating the effect of GnRH on the synthesis and secretion of gonadotropin hormone which acts on gonad to stimulate gametogenesis, gonadal cell proliferation, and the production of gonadal steroids [1]
Our results suggest that HdhGnRH-R gene synthesized in the neural ganglia might be involved in the control of gonadal maturation and gametogenesis of H. discus hannai
The GnRH-R superfamily consists of four G protein-coupled receptor family (GPCR) members: invertebrate GnRH receptor, corazonin receptor (Crz-R), adipokinetic hormone receptor (AKH-R), and AKH/Crz-related peptide receptor (ACP-R) [6]
Summary
Gonadotropin-releasing hormone receptors (GnRH-Rs) play crucial roles in mediating the effect of GnRH on the synthesis and secretion of gonadotropin hormone which acts on gonad to stimulate gametogenesis, gonadal cell proliferation, and the production of gonadal steroids [1]. GnRH-Rs are localized in anterior pituitary gonadotrophs/or peripheral tissues of mammalian and non-mammalian vertebrates [3]. These receptors can mediate their actions by association with G-proteins that can activate a phosphatidylinositol-calcium second messenger system to initiate cellular signaling pathways to increase the synthesis and secretion of luteinizing hormone. The GnRH-R superfamily consists of four G protein-coupled receptor family (GPCR) members: invertebrate GnRH receptor (invGnRH-R), corazonin receptor (Crz-R), adipokinetic hormone receptor (AKH-R), and AKH/Crz-related peptide receptor (ACP-R) [6]. GnRH-R superfamily in invertebrates has evolved by gene duplication as follows: amphioxus and ancestral species of protostomes and deuterostomes possess both vertebrate and invGnRH-R, mollusks, and annelids possess both invGNRH-R and AKH-R, and arthropods possess
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