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BOR - Papers in Press, published online ahead of print August 20, 2008.
Biol Reprod 2008, 10.1095/biolreprod.108.069690
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BIOLOGY OF REPRODUCTION 79, 1030–1037 (2008)
DOI: 10.1095/biolreprod.108.069690
© 2008 by the Society for the Study of Reproduction, Inc.

Hormonal Regulation of MicroRNA Expression in Periovulatory Mouse Mural Granulosa Cells1

Stephanie D. Fiedler , Martha Z. Carletti , Xiaoman Hong , and Lane K. Christenson 2

Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas 66160

ABSTRACT

MicroRNAs (miRNAs) mediate posttranscriptional gene regulation by binding to the 3' untranslated region of messenger RNAs to either inhibit or enhance translation. The extent and hormonal regulation of miRNA expression by ovarian granulosa cells and their role in ovulation and luteinization is unknown. In the present study, miRNA array analysis was used to identify 212 mature miRNAs as expressed and 13 as differentially expressed in periovulatory granulosa cells collected before and after an ovulatory dose of hCG. Two miRNAs, Mirn132 and Mirn212 (also known as miR-132 and miR-212), were found to be highly upregulated following LH/hCG induction and were further analyzed. In vivo and in vitro temporal expression analysis by quantitative RT-PCR confirmed that LH/hCG and cAMP, respectively, increased transcription of the precursor transcript as well as the mature miRNAs. Locked nucleic acid oligonucleotides complementary to Mirn132 and Mirn212 were shown to block cAMP-mediated mature miRNA expression and function. Computational analyses indicated that 77 putative mRNA targets of Mirn132 and Mirn212 were expressed in ovarian granulosa cells. Furthermore, upon knockdown of Mirn132 and Mirn212, a known target of Mirn132, C-terminal binding protein 1, showed decreased protein levels but no change in mRNA levels. The following studies are the first to describe the extent of miRNA expression within ovarian granulosa cells and the first to demonstrate that LH/hCG regulates the expression of select miRNAs, which affect posttranscriptional gene regulation within these cells.

corpus luteum, granulosa cells, luteinizing hormone, microRNA, ovulation

FOOTNOTES

1Supported by National Institutes of Health grant HD051870-01.

Correspondence: 2Lane K. Christenson, 3075 KLSIC, 3901 Rainbow Blvd., Kansas City, KS 66160. FAX: 913 588 7180; e-mail: lchristenson{at}kumc.edu




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M. Z Carletti and L. K Christenson
Rapid effects of LH on gene expression in the mural granulosa cells of mouse periovulatory follicles
Reproduction, May 1, 2009; 137(5): 843 - 855.
[Abstract] [Full Text] [PDF]


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