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This version published online on August 12, 2004
Endocrinology, doi:10.1210/en.2004-0710
A more recent version of this article appeared on November 1, 2004
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Submitted on June 3, 2004
Accepted on August 4, 2004

Cytochrome P450-dependent Lipid Metabolism in Pre-ovulatory Follicles

J. W. Newman, J. E. Stok, J. D. Vidal, C. J. Corbin, Q. Huang, B. D. Hammock, and A. J. Conley*

Department of Entomology, UC Davis Cancer Research Centerand Department of Population Health & Reproduction, University of California, Davis CA, 95616

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

Estrogen biosynthesis and proteolysis are both important processes involved in ovarian follicular development, which may be influenced by cytochrome P450s (CYPs)-dependent fatty acid metabolites. However, CYP-dependent lipid metabolism has not been characterized with respect to follicular maturation in vivo. Therefore, follicular fluid (FF) was collected in the hours before and after the LH surge in pigs, and concentrations of epoxy, hydroxy, and dihydroxy lipids were measured by LC/MS/MS. Arachidonate oxidation and epoxyeicosatrienoic acid (EET) hydrolysis to dihydroxyeicosatrienoic acids (DHETs) were also assessed in theca and granulosa tissue fractions, and the expression of CYP epoxygenases were evaluated by immunoblots using available anti-sera. To evaluate soluble epoxide hydrolase (sEH) expression, the porcine sEH was cloned from ovarian tissue, expressed and purified for antibody generation. The FF oxylipin concentrations ranged from 1-150 nM depending on compound and estrous stage. The FF concentrations of CYP-dependent oxylipins increased at estrus, as did sEH expression, however significant changes in epoxides were not observed and the 11,12-DHET peak was delayed. The ratio of 14,15-:11,12-DHET across all samples correlated with the log of FF estradiol concentrations (P < 0.01). Epoxygenase activities were similar in theca and granulosa, varying little with follicular development, while the decline of a single CYP2J isoform at ovulation was observed by immunoblots. The sEH activity was higher in granulosa than theca. Finally, the dynamic changes in follicular CYP-dependent arachidonic acid metabolites and their modulatory function in vascular models suggest roles for these metabolites in follicular maturation, which may include regulation of estradiol biosynthesis and pre-ovulatory remodeling of the follicular wall that should be fully explored in future studies.




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