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BOR - Papers in Press, published online ahead of print November 24, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.035949
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Submitted September 2, 2004
Returned for revision October 13, 2004
Accepted November 8, 2004

Pregnancy


Expression of Estrogen Receptors-{alpha} and -{beta} in the Pregnant Ovine Uterine Artery Endothelial Cells In Vivo and In Vitro

Wu Xiang Liao , Ronald R. Magness , and Dong-bao Chen *

* To whom correspondence should be addressed. E-mail: dochen{at}ucsd.edu.

Abstract
Estrogen is recognized to be one of the driving forces to increase uterine blood flow through both rapid and delayed actions via binding to its receptor(s) (ER{alpha}and/or ER{beta}) at the uterine artery (UA) wall especially the endothelium (UAE). However, the information regarding ER expression in UAE is limited. This study is designed to test if ER(s) are expressed in UAE in vivo, and if so, are these receptors maintained in cultured UA endothelial cells (UAEC) in vitro? By using immunohistochemical and Western blot analyses, we clearly demonstrated ER{alpha} and ER{beta} protein expression in pregnant (D 120-130) sheep UA and UAE in vivo and as well as cultured UAEC in vitro. RT-PCR amplified both ER{alpha} and ER{beta} mRNAs in UA, UAE and UAEC. Interestingly, a truncated ER{beta} (ER{beta}2) variant due to splicing deletion of exon 5 of ER{beta} gene was detected in these cells. Quantitative RT-PCR analysis revealed that ER{alpha} mRNA level is ~8-fold (P < 0.01) higher than that of ER{beta} in UAEC, indicating that ER{alpha} may play a more important role than ER{beta} in UAEC responses to estrogen. Fluorescence immunolabeling analysis showed that ER{alpha} is present in both nucleus and plasma membrane in UAEC, and the later is also co-localized with caveolin-1. The membrane and nuclear ER{alpha} presumably participate in rapid and delayed responses to estrogen on UAE, respectively. Taken together, our data demonstrated that UAE is a direct target of estrogen actions and the UAEC culture model established is suitable for dissecting estrogen actions on UAE.

Key words: Mechanisms of Hormone Action • Pregnancy • Estradiol receptor • Nitric oxide • Uterus


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