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Complementary Deoxyribonucleic Acid Cloning of Spermatogonial Stem Cell Renewal Factor

Laboratory of Fish Reproductive Physiology (T.M.), Faculty of Agriculture, Ehime University, Matsuyama 790-8566, Japan; "Time’s Arrow and Biosignaling" PRESTO Japan Science and Technology Agency (T.M., C.I.M.), Kawaguchi 332-0012, Japan; and Faculty of Fisheries (T.O., K.Y.), Hokkaido University, Hakodate 041-8611, Japan

Address all correspondence and requests for reprints to: Takeshi Miura, Laboratory of Fish Reproductive Physiology, Faculty of Agriculture, Ehime University, 3-5-7 Tarumi, Matsuyama 790-8566, Japan. E-mail: miutake{at}agr.ehimeu.ac.jp.

Spermatogonial mitosis can be subdivided into two processes: spermatogonial stem cell renewal and spermatogonial proliferation toward meiosis. Recently it has been indicated that estrogen, estradiol-17ß, is involved in regulating the renewal of spermatogonial stem cells in eel. To determine the genes that directly regulate this process, we used expression screening to identify genes whose expression is regulated by estradiol-17ß in testes. We detected a previously unidentified cDNA clone that is up-regulated by estradiol-17ß stimulation and named it eel spermatogenesis-related substances 34 (eSRS34) cDNA. Homology searching showed that eSRS34 shares amino acid sequence similarity with human platelet-derived endothelial cell growth factor. We examined the function of eSRS34 using several in vitro systems. Recombinant eSRS34 produced by a baculovirus system induced spermatogonial mitosis in testicular organ culture. Furthermore, the addition of an antibody specific for eSRS34 prevented spermatogonial mitosis induced by estradiol-17ß stimulation in a germ cell/somatic cell coculture system. We therefore conclude that eSRS34 is a "spermatogonial stem cell renewal factor."

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