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Hum. Reprod. Advance Access originally published online on February 9, 2007
Human Reproduction 2007 22(5):1247-1252; doi:10.1093/humrep/del519
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© The Author 2007. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Regulation of granulosa cell proliferation and EGF-like ligands during the periovulatory interval in monkeys

K.N. Fru1, M. Cherian-Shaw2, M. Puttabyatappa2, C.A. VandeVoort3 and C.L. Chaffin2,4

1 Department of Physiology, Medical College of Georgia, Augusta, GA, USA 2 Department of Obstetrics, Gynecology and Reproductive Sciences, University of Maryland School of Medicine, Baltimore, MD, USA 3 California National Primate Research Center, Davis, CA, USA

4 To whom correspondence should be addressed at: Department of Obstetrics, Gynecology and Reproductive Sciences, University of Maryland School of Medicine, Baltimore, MD, 21201, USA. E-mail: cchaffin{at}upi.umaryland.edu

BACKGROUND: This study seeks to clarify cell cycle dynamics of granulosa cells following hCG and elucidate the expression of epidermal growth factor (EGF)-like ligands during luteinization.

METHODS: Granulosa cells were obtained from rhesus macaques undergoing controlled ovarian stimulation protocols before or after an ovulatory hCG bolus. Cell cycle characteristics were determined by flow cytometry and levels of EGF receptor (EGFR), amphiregulin (AREG), epiregulin (EREG) and betacellulin (BTC) mRNAs were measured by real-time RT-PCR.

RESULTS: The proportion of cells in S-phase was 7.5% prior to hCG and did not decline until 24 h after hCG (3.1%). EGFR protein and BTC mRNA did not change following hCG, whereas AREG and EREG mRNA increased starting at 3 and 12 h post-hCG, respectively, and remained elevated thereafter.

CONCLUSIONS: Cell cycle transit of macaque granulosa cells does not change until 24 h after an ovulatory stimulus, whereas the EGF-like ligands EREG and AREG are increased rapidly. This suggests that luteinizing granulosa cells are refractory to mitogenic stimulation by EGFR ligands.

Key words: non-human primate/granulosa cell/proliferation/luteinization/epidermal growth factor

Submitted on August 16, 2006; resubmitted on October 25, 2006; resubmitted on November 14, 2006; accepted on November 29, 2006.


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