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J. Biol. Chem., Vol. 280, Issue 9, 7685-7693, March 4, 2005

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Identification and Integrative Analysis of 28 Novel Genes Specifically Expressed and Developmentally Regulated in Murine Spermatogenic Cells^*

Sungeun Hong, Inchul Choi, Jong-Min Woo, Jungsu Oh, Taewan Kim, Eunyoung Choi, Tae-Wan Kim, Yong-Keun Jung, Do Han Kim, Choong-Hyun Sun, Gwan-Su Yi, Edward M. Eddy¶, and Chunghee Cho||

From the Department of Life Science, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea, the School of Engineering, Information and Communications University, Daejon 305-714, Korea, and the ^¶Gamete Biology Section, Laboratory of Reproductive and Developmental Toxicology, NIEHS, National Institutes of Health, Research Triangle Park, North Carolina 27709

Mammalian spermatogenesis is a highly ordered process that occurs in mitotic, meiotic, and postmeiotic phases. The unique mechanisms responsible for this tightly regulated developmental process suggest the presence of an intrinsic genetic program composed of spermatogenic cell-specific genes. In this study, we analyzed the mouse round spermatid UniGene library currently containing 2124 gene-oriented transcript clusters, predicting that 467 of them are testis-specific genes, and systematically identified 28 novel genes with evident testis-specific expression by in silico and in vitro approaches. We analyzed these genes by Northern blot hybridization and cDNA cloning, demonstrating the presence of additional transcript sequences in five genes and multiple transcript isoforms in six genes. Genomic analysis revealed lack of human orthologues for 10 genes, implying a relationship between these genes and male reproduction unique to mouse. We found that all of the novel genes are expressed in developmentally regulated and stage-specific patterns, suggesting that they are primary regulators of male germ cell development. Using computational bioinformatics tools, we found that 20 gene products are potentially involved in various processes during spermatogenesis or fertilization. Taken together, we predict that over 20% of the genes from the round spermatid library are testis-specific, have discovered the 28 authentic, novel genes with probable spermatogenic cell-specific expression by the integrative approach, and provide new and thorough information about the novel genes by various in vitro and in silico analyses. Thus, the study establishes on a comprehensive scale a new basis for studies to uncover molecular mechanisms underlying the reproductive process.

Received for publication, November 3, 2004 , and in revised form, December 21, 2004.

^* This work was supported by Korea Research Foundation Grant KRF-2002-070-C0007, Korea Ministry of Science and Technology Systems Biology Research Grant M1–0309-00-0006, and the Korea Ministry of Education Brain Korea 21 project. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains four additional tables.

^|| To whom correspondence should be addressed. Tel.: 82-62-970-2490; Fax: 82-62-970-2484; E-mail: choch{at}gist.ac.kr.

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