cDNA cloning of GTH receptor family in starfish ovaries

In starfish, oocytes in the ripe ovary do not develop beyond the end of the first prophase stage of meiosis. Such immature oocytes are not fertilisable. The resumption of meiosis in these oocytes and release from the ovary are induced by the maturation-indueing hormone 1-methyladenine (1-MeAde) (Kanatani et al., 1969). 1-MeAde is produced by ovarian follicle cells after stimulation by a gonad-stimulating hormone (GSS) secreted from radial nerves. The action of GSS on 1-MeAde production in follicle cells appears to be mediated by its receptor, G-proteins and adenylyl cyclase (Mita & Nagahama, 1991). It has been shown that upon incubation of follicle cells with GSS, there is a dose-related increase in cyclic AMP production, coincident with an increase in 1-MeAde production (Mita et al., 1989). Thus, it was suggested that a G-protein coupled (seven transmembrane type) receptor is involved in GSS signal transduction in a similar manner to the pituitary–gonadal axis in vertebrates. To elucidate the mechanism of starfish oocyte maturation, we focused on a receptor protein for GSS on the ovarian follicle cell surface. In this study, we cloned the cDNA with glycoprotein hormone receptor homology in a transmembrane region from ovaries of the starfish Asterina pectinifera.Using consensus sequences from glycoprotein hormone receptors, a 3953 bp cDNA was cloned from mRNA of starfish A. pectinifera ovaries. The region from nucleotide 72 to 3109 represents the open-reading frame. The seven transmembrane spanning domains were found in the region from nucleotide 1745 to 2533. The lengths of specific domains in the cDNA were 1671 bp, 789 bp and 579 bp for extracellular (ECD), transmembrane (TMD) and intracellular domains (ICD), respectively. Similar to other glycoprotein hormone receptors, the cDNA from starfish ovaries had a large ECD region. With respect to the predicted amino acid sequence, the ECD, TMD and ICD were composed of 557, 263 and 192 amino acids, respectively.