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*CHORIONIC GONADOTROPIN
Molecular Endocrinology 11 (2): 172-182
Copyright © 1997 by The Endocrine Society

Estrogen Receptor-ß mRNA Expression in Rat Ovary: Down-Regulation by Gonadotropins

Michael Byers, George G. J. M. Kuiper, Jan-Åke Gustafsson and Ok-Kyong Park-Sarge

Department of Physiology (M.B., O-K.P-S.) University of Kentucky Lexington, Kentucky 40536
Center for Biotechnology (G.G.J.M.K.) and Department of Medical Nutrition (J-Å.G.) Karolinska Institute S-14186 Huddinge, Sweden

We have examined the expression and regulation of the two estrogen receptor (ER{alpha} and ERß) genes in the rat ovary, using Northern blotting, RT-PCR, and in situ hybridization histochemistry. Northern blotting results show that the ovary expresses both ER{alpha} and ERß genes as single (~6.5-kb) and multiple (ranging from ~1.0-kb to ~10.0-kb) transcripts, respectively. ER{alpha} mRNA is expressed at a level lower than ERß mRNA in immature rat ovaries. This relationship appears unchanged between sexually mature adult rats and immature rats. In sexually mature adult rats undergoing endogenous hormonal changes, whole ovarian content of ERß mRNA, as determined by RT-PCR, remained more or less constant with the exception of the evening of proestrus when ERß mRNA levels were decreased. Examination of ERß mRNA expression at the cellular level, by in situ hybridization, showed that ERß mRNA is expressed preferentially in granulosa cells of small, growing, and preovulatory follicles, although weak expression of ERß mRNA was observed in a subset of corpora lutea, and that the decrease in ERß mRNA during proestrous evening is attributable, at least in part, to down-regulation of ERß mRNA in the preovulatory follicles. This type of expression and regulation was not typical for ER{alpha} mRNA in the ovary. Although whole ovarian content of ER{alpha} mRNA was clearly detected by RT-PCR, no apparent modulation of ER{alpha} mRNA levels was observed during the estrous cycle. Examination of ER{alpha} mRNA expression at the cellular level, by in situ hybridization, showed that ER{alpha} mRNA is expressed at a low level throughout the ovary with no particular cellular localization.

To further examine the potential role of the preovulatory pituitary gonadotropins in regulating ERß mRNA expression in the ovary, we used immature rats treated with gonadotropins. In rats undergoing exogenous hormonal challenges, whole ovarian content of ERß mRNA, as determined by RT-PCR, remained more or less unchanged after an injection of PMSG. In contrast, a subsequent injection of human CG (hCG) resulted in a substantial decrease in whole ovarian content of ERß mRNA. In situ hybridization for ERß mRNA shows that small, growing, and preovulatory follicles express ERß mRNA in the granulosa cells. The preovulatory follicles contain ERß mRNA at a level lower than that observed for small and growing follicles. In addition, there is an abrupt decrease in ERß mRNA expression in the preovulatory follicles after hCG injection. The inhibitory effect of hCG on ERß mRNA expression was also observed in cultured granulosa cells. Moreover, agents stimulating LH/CG receptor-associated intracellular signaling pathways (forskolin and a phorbol ester) readily mimicked the effect of hCG in down-regulating ERß mRNA in cultured granulosa cells.

Taken together, our results demonstrate that 1) the ovary expresses both ER{alpha} and ERß genes, although ERß is the predominant form of estrogen receptor in the ovary, 2) ERß mRNA is localized predominantly to the granulosa cells of small, growing, and preovulatory follicles, and 3) the preovulatory LH surge down-regulates ERß mRNA. These results clearly implicate the physiological importance of ERß in female reproductive functions.




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