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BOR - Papers in Press, published online ahead of print December 15, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.035899
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Submitted September 3, 2004
Returned for revision October 5, 2004
Accepted November 16, 2004

Ovary


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

Jon D. Hennebold *, Kuni Mah , Wilma Perez , Jessica E. Vance , Richard L. Stouffer , Christophe Morisseau , Bruce D. Hammock , and Eli Y. Adashi

* To whom correspondence should be addressed. E-mail: henneboj{at}ohsu.edu.

Abstract
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 pre-ovulatory follicles. Ephx2A mRNA expression, however, was detectable in follicles at different stages of development, as well as in the corpus luteum (CL). Total ovarian epoxide hydrolase activity increased following the induction of follicular development, and remained elevated through the periovulatory and post-ovulatory 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.

Key words: Ovary • Corpus luteum • Granulosa cells • Ovulation


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