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Regular Article |
a Department of Animal Sciences, The Ohio State University, Columbus, Ohio 43210
b Department of Obstetrics and Gynecology, Indiana University School of Medicine, Indianapolis, Indiana 46202
c Medical Sciences, Indiana University, Bloomington, Indiana 47405
ABSTRACT
Objectives were to sequence and examine the expression of the estrogen receptor ß (ERß) in the sheep ovary. The sequence of the ovine ERß (oERß) was determined using reverse-transcription polymerase chain reaction (RT-PCR) and cloning techniques. The reading frame of oERß contained 527 amino acids and exhibited high overall homology with cow (98%), rat (88%), and human (88%) ERß. In addition, an oERß isoform having a 139-base pair deletion (oERß1) was identified. The predicted amino acid sequence of this isoform is lacking the ligand-binding and carboxyl-terminal transactivation domains. The oERß protein and mRNA were determined in ovaries obtained from ewes on Days 0 (first day of estrus), 2, 6, and 10 of the estrous cycle and Day 30 of gestation. Immunohistochemistry showed that oERß protein was located in granulosa cells, the ovarian surface epithelium, endothelium, and Day 2 corpus luteum (CL). Weak immunostaining for ERß was detected in the theca interna. Relative steady-state amounts of oERß mRNA in the CL were determined using semiquantitative RT-PCR. Amounts of oERß mRNA were greater (P < 0.05) during CL formation (Day 2) than at later stages. The oERß to oERß1 mRNA ratio was lower (P < 0.05) on Day 2 than on Day 10 or Day 30 due to a decrease in amounts of oERß1. Results indicate that the oERß is a 527-amino acid protein expressed in specific cells of the ovary. Changes in relative amounts of full-length oERB and a deletion isoform in CL occurred during the estrous cycle, suggesting that these two types of ERß might regulate estrogen actions during early CL development in sheep.
First decision: 9 January 2001.
1 Salaries and research support provided by State and Federal funds appropriated to The Ohio Agricultural Research and Development Center and The Ohio State University. This research is manuscript no. 42-00AS and was supported by funds from the National Institutes of Health, PHS CA74748 to K.P.N. and CA74748 postdoctoral award to H.C.
2 Correspondence: H. Cárdenas, Department of Animal Sciences, The Ohio State University, 2029 Coffey Rd., Columbus, OH 43210. FAX: 614 292 7116; cardenas-seijas.2{at}osu.edu
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