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Identification and Characterization of Two Distinct GnRH Receptor Subtypes in a Teleost, the Medaka Oryzias latipes

Department of Aquatic Bioscience (K.O., Y.Y., K.A.), Graduate School of Agricultural and Life Sciences, and Department of Integrated Biosciences (S.N., R.K., H.K.), Graduate School of Frontier Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan; and Department of Integrated Biosciences (H.M., M.K., A.S.), Graduate School of Frontier Sciences, The University of Tokyo, and Department of Biological Sciences (K.N.), Graduate School of Science, The University of Tokyo, Bunkyo, Tokyo 113-0033, Japan

Address all correspondence and requests for reprints to: Dr. K. Aida, Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan. E-mail: aida{at}uf.a.u-tokyo.ac.jp

We report the identification and characterization of two distinct GnRH receptor (GnRH-R) subtypes, designated GnRH-R1 and GnRH-R2, in a model teleost, the medaka Oryzias latipes. These seven-transmembrane receptors of the medaka contain a cytoplasmic C-terminal tail, which has been found in all other nonmammalian GnRH-Rs cloned to date. The GnRH-R1 gene is composed of three exons separated by two introns, whereas the GnRH-R2 gene has an additional intron and therefore consists of four exons and three introns. The GnRH-R1 and GnRH-R2 genes, both of which exist as single-copy genes in the medaka genome, were mapped to linkage groups 3 and 16, respectively. Inositol phosphate assays using COS-7 cells transfected with GnRH-R1 and GnRH-R2 demonstrated that they had remarkably different ligand sensitivities, although both receptors showed highest preference for chicken-II-type GnRH. Phylogenetic analysis showed the presence of three paralogous lineages for vertebrate GnRH-Rs and indicated that neither GnRH-R1 nor GnRH-R2 is the medaka ortholog to mammalian GnRH-Rs that lack a cytoplasmic tail. This, together with an observation that medaka-type GnRH had low affinity for GnRH-R1 and GnRH-R2, suggests that a third GnRH-R may exist in the medaka.

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