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Expression of Estrogen Receptors- and -ß in the Pregnant Ovine Uterine Artery Endothelial Cells In Vivo and In Vitro¹

Department of Reproductive Medicine,³ University of California San Diego, La Jolla, California 92093-0802 Perinatal Research Laboratories, Departments of Obstetrics and Gynecology,⁴ University of Wisconsin-Madison, Meriter Hospital, Madison, Wisconsin 53715

Estrogen is recognized to be one of the driving forces in increases in uterine blood flow through both rapid and delayed actions via binding to its receptors, ER and ERß at the uterine artery (UA) wall, and especially in UA endothelium (UAE). However, information regarding estrogen receptor (ER) expression in UAE is limited. This study was designed to test whether ERs are expressed in UAE in vivo, and if they are, whether these receptors are maintained in cultured UA endothelial cells (UAECs) in vitro. By using immunohistochemical and Western blot analyses, we clearly demonstrated ER and ERß protein expression in pregnant (Days 120–130) sheep UA and UAE in vivo and as well as cultured UAECs in vitro. Reverse transcription-polymerase chain reaction (RT-PCR) amplified both ER and ERß mRNAs in UA, UAE, and UAECs. Of interest, a truncated ERß (ERß2) variant due to a splicing deletion of exon 5 of the ERß gene was detected in these cells. Quantitative RT-PCR analysis revealed that ER mRNA levels are 8-fold (P < 0.01) higher than that of ERß in UAECs, indicating that ER may play a more important role than ERß in the UAEC responses to estrogen. Fluorescence immunolabeling analysis showed that ER is present in both nuclei and plasma membranes in UAECs, and the latter is also colocalized with caveolin-1. The membrane and nuclear ER presumably participate in rapid and delayed responses, respectively, to estrogen on UAE. Taken together, our data demonstrated that UAE is a direct target of estrogen actions and that the UAEC culture model we established is suitable for dissecting estrogen actions on UAE.

¹ Supported in part by National Institutes of Health (NIH) RO1 grants HL70562 and HL74947, by an Academic Senate grant from the University of California San Diego to D.B.C., and by NIH RO1 grants HD33255 and HL49210 to R.R.M.

² Correspondence: Dong-bao Chen, Division of Maternal-Fetal Medicine (MC0802), Department of Reproductive Medicine, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0802. FAX 619 543 2919; dochen{at}ucsd.edu

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