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Identification and Characterization of an Ovary-Selective Isoform of Epoxide Hydrolase¹

Division of Reproductive Sciences,³ Oregon National Primate Research Center, Beaverton, Oregon 97006 Department of Obstetrics and Gynecology,⁴ Oregon Health & Science University, Portland, Oregon 97239 Department of Entomology,⁵ University of California-Davis, Davis, California 95616 Department of Obstetrics and Gynecology,⁶ University of Utah Health Sciences Center, Salt Lake City, Utah 84132

A novel ovary-selective gene was identified by suppression subtractive hybridization (SSH) that is expressed only during the mouse periovulatory phase of a stimulated estrous cycle. Analysis of the protein encoded by the full-length cDNA revealed that the majority of it, with the exception of the first 44 amino acids, matched soluble epoxide hydrolase (Ephx2, referred to as Ephx2A). By comparing the cDNA sequence of this newly identified variant of soluble epoxide hydrolase (referred to as Ephx2B) with the mouse genome database, an exon was identified that corresponds to its unique 5' cDNA sequence. Through the use of an Ephx2A-specific probe, Northern blot analysis revealed that this mRNA was also expressed in the ovary, with the highest level of expression occurring during the luteal phase of a stimulated estrous cycle. In situ hybridization revealed that Ephx2B mRNA expression was restricted to granulosa cells of preovulatory follicles. Ephx2A mRNA expression, however, was detectable in follicles at different stages of development, as well as in the corpus luteum. Total ovarian epoxide hydrolase activity increased following the induction of follicular development, and remained elevated through the periovulatory and postovulatory stages of a stimulated estrous cycle. The change in enzyme activity paralleled the combined mRNA expression profiles for both Ephx2A and Ephx2B, thus supporting a role for epoxide metabolism in ovarian function.

¹ Supported in part by National Institutes of Health grants HD42000 (to J.D.H.), NCRR00163 (to J.D.H. and R.L.S.), HD30288 and 37845(to E.Y.A.), HD20869 (to R.L.S.), HD19182 (to K.M.), and by grants ES02710 and ES04699 from the National Institute of Environmental Health Sciences (to C.M. and B.D.H.).

² Correspondence: Jon D. Hennebold, Division of Reproductive Sciences, Oregon National Primate Research Center, Oregon Health & Science University, 505 NW 185th Ave., Beaverton, OR 97006. FAX: 503 690-5563; henneboj{at}ohsu.edu

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