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Teleost Ovarian Carbonyl Reductase-Like 20ß-Hydroxysteroid Dehydrogenase: Potential Role in the Production of Maturation-Inducing Hormone During Final Oocyte Maturation¹

^a Laboratory of Reproductive Biology, National Institute for Basic Biology, Okazaki 444-8585, Japan ^b Division of Biological Science, Graduate School of Science, Hokkaido University, Kita-Ku, Sapporo 060-0810, Japan ^c Department of Biochemistry, Faculty of Pharmaceutical Sciences, Hoshi University, Shinagawa-Ku, Tokyo 142-0063, Japan ^d Division of Gene Expression and Regulation I, National Institute for Basic Biology, Okazaki 444-8585, Japan ^e Sado Marine Biological Station, Niigata University, Niigata 952-2135, Japan ^f CREST, Japan Science Technology Corporation, Kawaguchi, Saitama 332-0012, Japan

17,20ß-Dihydroxy-4-pregnen-3-one is the major oocyte maturation-inducing hormone of several teleost species. Gonadotropin-induced increase in ovarian 20ß-hydroxysteroid dehydrogenase activity is essential for the synthesis of maturation-inducing hormone. Cloning and expression studies suggest that ayu (Plecoglossus altivelis) ovarian carbonyl reductase can function as 20ß-hydroxysteroid dehydrogenase. The amino acid sequence deduced from the isolated cDNA had 276 amino acid residues and shared approximately 60% homology with mammalian and teleostean carbonyl reductases. The sequence data search showed that the ayu cDNA clone belongs to the short-chain dehydrogenase/reductase family. The clear lysate prepared from Escherichia coli harboring the cDNA catalyzed the production of maturation-inducing hormone. Its identification was confirmed by two-dimensional, thin-layer chromatography followed by recrystallization. Purification of the E. coli-expressed cDNA product revealed that it possessed both carbonyl reductase and steroid dehydrogenase activities, and 17-hydroxyprogesterone, the endogenous immediate precursor of maturation-inducing hormone, was one of the preferred substrates. Furthermore, Northern blot analysis denoted that the transcripts are present both in fully grown, immature ovarian follicles and at higher levels in mature ovarian follicles. These results demonstrate that the carbonyl reductase of ayu ovary is involved in the production of maturation-inducing hormone, and they provide evidence for a novel physiological role of this enzyme in the final maturation of oocytes. Based on its functional properties, the enzyme can be referred to as carbonyl reductase-like 20ß-hydroxysteroid dehydrogenase.

First decision: 28 November 2001.

¹ Supported in part by Grants-in-Aid from the Japan Society for the Promotion of Science (JSPS-RFTF 96L00401 to Y.N.); Scientific Research (07283103, 08454266, and 10440247 to Y.N. and 08740642 to M.T.) from the Ministry of Education, Science, Culture, and Sports, Japan, and CREST of Japan Science Technology Corporation; and a Grant-in-Aid (Bio Media Program) from the Ministry of Agriculture, Forestry, and Fisheries.

² Correspondence. FAX: 81 564 55 7556; nagahama{at}nibb.ac.jp

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