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Endocrinology Vol. 141, No. 5 1764-1771
Copyright © 2000 by The Endocrine Society

ARTICLES

Complementary Deoxyribonucleic Acid Cloning, Gene Expression, and Ligand Selectivity of a Novel Gonadotropin-Releasing Hormone Receptor Expressed in the Pituitary and Midbrain of Xenopus laevis1

B. E. Troskie, J. P. Hapgood, R. P. Millar and N. Illing

Medical Research Council Research Unit for Molecular Reproductive Endocrinology (B.E.T.), Department of Medical Biochemistry, University of Cape Town, Observatory 7925; Medical Research Council Reproductive Biology Unit (R.P.M.), Center for Reproductive Biology, Edinburgh, United Kingdom EH3 9ET; Department of Biochemistry (J.P.H)., University of Stellenbosch, Matieland 7602; and Department of Biochemistry (N.I.), University of Cape Town, Rondebosch 7700, South Africa

Address all correspondence and requests for reprints to: Dr. B. E. Troskie, Medical Research Council Research Unit for Molecular Reproductive Endocrinology, Department of Medical Biochemistry, University of Cape Town, Observatory 7925, South Africa. E-mail: Brigitte{at}Curie.uct.ac.za

We have cloned the full-length complementary DNA (cDNA) for a GnRH receptor from Xenopus laevis pituitary cDNA and determined its gene structure. The cDNA encodes a 368-amino acid protein that has a 46% amino acid identity to the human GnRH receptor. The X. laevis GnRH receptor has all of the amino acids identified in the mammalian GnRH receptors as sites of interaction with the GnRH ligand. However, this receptor cDNA shares the same distinguishing structural features of the GnRH receptor that have been characterized from other nonmammalian vertebrates. These include the pair of aspartate residues in the transmembrane domains II and VII compared with the aspartate/asparagine arrangement in mammalian receptors, the amino acid PEY motif in extracellular loop III (SEP in mammals), and the presence of a carboxyl-terminal tail. Previous studies have reported that mammalian GnRH was equipotent to other naturally occurring GnRH subtypes in stimulating LH release from the amphibian pituitary. However, in this study we show that the X. laevis GnRH receptor has ligand selectivity for the naturally occurring GnRHs similar to other nonmammalian GnRH receptors. The order of potency of the GnRHs in stimulating inositol phosphate production in COS-1 cells transiently transfected with the X. laevis GnRH receptor cDNA was chicken GnRH II>salmon GnRH>mammalian GnRH. Transcripts of this GnRH receptor are expressed in the pituitary and midbrain of X. laevis.




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