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BOR - Papers in Press, published online ahead of print August 30, 2006.
Biol Reprod 2006, 10.1095/biolreprod.106.053769
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BIOLOGY OF REPRODUCTION 75, 868–876 (2006)
DOI: 10.1095/biolreprod.106.053769
© 2006 by the Society for the Study of Reproduction, Inc.

research-article

Primate Granulosa Cell Response via Prostaglandin E2 Receptors Increases Late in the Periovulatory Interval1

Nune Markosyan 2, Brandy L. Dozier , Frank A. Lattanzio , and Diane M. Duffy 

Department of Physiological Sciences, Eastern Virginia Medical School, Norfolk, Virginia 23507-1980

ABSTRACT

Successful ovulation requires elevated follicular prostaglandin E2 (PGE2) levels. To determine which PGE2 receptors are available to mediate periovulatory events in follicles, granulosa cells and whole ovaries were collected from monkeys before (0 h) and after administration of an ovulatory dose of hCG to span the 40-h periovulatory interval. All PGE2 receptor mRNAs were present in monkey granulosa cells. As assessed by immunofluorescence, PTGER1 (EP1) protein was low/nondetectable in granulosa cells 0, 12, and 24 h after hCG but was abundant 36 h after hCG administration. PTGER2 (EP2) and PTGER3 (EP3) proteins were detected by immunofluorescence in granulosa cells throughout the periovulatory interval, and Western blotting showed an increase in PTGER2 and PTGER3 levels between 0 h and 36 h after hCG. In contrast, PTGER4 (EP4) protein was not detected in monkey granulosa cells. Granulosa cell response to PGE2 receptor agonists was examined 24 h and 36 h after hCG administration, when elevated PGE2 levels present in periovulatory follicles initiate ovulatory events. PGE2 acts via PTGER1 to increase intracellular calcium. PGE2 increased intracellular calcium in granulosa cells obtained 36 h, but not 24 h, after hCG; this effect of PGE2 was blocked by a PTGER1 antagonist. A PTGER2-specific agonist and a PTGER3-specific agonist each elevated cAMP in granulosa cells obtained 36 h, but not 24 h, after hCG. Therefore, the granulosa cells of primate periovulatory follicles express multiple receptors for PGE2. Granulosa cells respond to agonist stimulation of each of these receptors 36 h, but not 24 h, after hCG, supporting the hypothesis that granulosa cells are most sensitive to PGE2 as follicular PGE2 levels peak, leading to maximal PGE2-mediated periovulatory effects just before ovulation.

granulosa cells, ovary, ovulation, signal transduction

FOOTNOTES

1Supported by National Institutes of Health grant HD39872 to D.M.D., and Eastern Virginia Medical School.

Correspondence: 2 Nune Markosyan, Department of Physiological Sciences, Eastern Virginia Medical School, 700 Olney Rd., Norfolk VA 23507-1980. FAX: 757 624 2269; e-mail: markosn{at}evms.edu






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Copyright © 2006 by the Society for the Study of Reproduction.