Identification, expression of a glycoprotein hormone receptor homolog in the amphioxus Branchiostoma belcheri with implications for origin of vertebrate GpHRs

Amphioxus is phylogenetically located at the most primitive position of the chordate clade. Its endocrine system has been extensively studied; however, key information about the glycoprotein hormone (GpH) and its receptor (GpHR) in the endocrine system remained to be elucidated. In this study, a GpHR homologous gene, BbGpHR-like, has been identified in the amphioxus Branchiostoma belcheri. It contains a 1377bp open reading frame that corresponds to a deduced protein of 458 amino acid residues with a predicted molecular mass of approximately 50kDa. Phylogenetic analysis shows that BbGpHR-like is located at the position between vertebrate and invertebrate, indicating amphioxus B. belcheri is in an evolutionary transitional state between vertebrate and invertebrate in the GpHR lineage. BbGpHR-like has been successfully expressed in Pichia pastoris. Western blot analysis confirmed that the receptor produced a cross-immunoreactivity with human thyrotropin receptor (TSHR), the luteinizing hormone receptor (LHR) and the follicle-stimulating hormone receptor (FSHR). In situ hybridization and immunohistochemical analyses revealed that the BbGpHR-like transcripts and proteins were strongly distributed in the ovaries, testes and/or endostyle. These findings suggest that BbGpHR-like possibly plays an ancient and fundamental role in the control of thyroid hormone synthesis and gametogenesis in chordates. Presumably, the amphioxus BbGpHR-like represents the ancestral form of the GpHR gene prior to its split to the vertebrate paralogs gonadotropin receptor and thyrotropin receptor.