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Biology of Reproduction 65, 269-276 (2001)
© 2001 Society for the Study of Reproduction, Inc.


Regular Article

DNA Array Analysis of Changes in Preovulatory Gene Expression in the Rat Ovary1

Chandra P. Leo3,a, Margareta D. Pisarskaa, and Aaron J.W. Hsueh2,a

a Division of Reproductive Biology, Department of Gynecology and Obstetrics, Stanford University Medical Center, Stanford, California 94305-5317

ABSTRACT

During the periovulatory period, the mammalian ovary is the site of dramatic functional and structural changes, leading to oocyte maturation, follicle rupture, and corpus luteum formation. To a large extent, these processes result from changes in the transcriptome of various ovarian cell types. To develop a broader view of periovulatory changes in gene expression in the ovary and to identify further genes involved in periovulatory events, we used the recently developed DNA array technology. Immature female eCG-primed rats were killed either immediately before or 6 h after ovulation induction with hCG. Total ovarian RNA was isolated and used to prepare radiolabeled cDNA probes, which were hybridized to DNA arrays representing approximately 600 rat genes. Quantitative analysis identified a multitude of regulated gene messages, including several genes involved in extracellular matrix degradation and lipid/steroid metabolism previously reported to be induced by hCG. This screening also identified a group of candidate genes whose ovarian expression and gonadotropin regulation was hitherto unknown. The induction of three of these genes, encoding cutaneous fatty acid-binding protein, the interleukin-4 receptor alpha chain, and prepronociceptin, was confirmed and further characterized by Northern blot analysis. In addition, in situ hybridization analysis showed that hCG administration resulted in exclusive or predominant expression of all three genes in theca cells. These results demonstrate that DNA arrays can be used to identify genes regulated during the periovulatory period, thus contributing to a more detailed understanding of the molecular mechanisms of ovulation.

FOOTNOTES

First decision: 26 December 2000.

1 This study was supported by NICHD/NIH through a cooperative agreement (U54-HD31398 to A.J.W.H.) as part of the Specialized Cooperative Centers Program in Reproduction Research, by the Core Facility of the Center, and by K12 HD01249 to M.D.P. C.P.L. is supported by a postdoctoral fellowship from the German Academic Exchange Service.

2 Correspondence: Aaron J.W. Hsueh, Division of Reproductive Biology, Department of Gynecology and Obstetrics, Stanford University Medical Center, 300 Pasteur Drive, Room A344, Stanford, CA 94305-5317. FAX: 650 725 7102; aaron.hsueh{at}stanford.edu

3 Current address: Department of Obstetrics and Gynecology, University of Leipzig, Leipzig, Germany.




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