Estrogen Receptor-Mediated Gene Expression in Cultured Epithelial Cells of Rat Efferent Ductules
Chen, Yu-Chyu
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Permalink
https://hdl.handle.net/2142/87135
Description
Title
Estrogen Receptor-Mediated Gene Expression in Cultured Epithelial Cells of Rat Efferent Ductules
Author(s)
Chen, Yu-Chyu
Issue Date
1998
Doctoral Committee Chair(s)
Hess, Rex A.
Department of Study
Veterinary Clinical Medicine
Discipline
Veterinary Clinical Medicine
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Molecular
Language
eng
Abstract
Estrogens have been detected in the rete testis fluid and semen. Estrogen receptors are localized in the efferent ductules and the head of the epididymis. However, knowledge of estrogen effects in the male reproductive tract is limited. In some species, concentrations of estrogens in testicular fluid are higher than in blood. A recent study in estrogen receptor knockout mice also showed an obvious morphological change in the efferent ductules. These past findings prompted the author to investigate the potential influence of estrogens on the epithelium lining the efferent ductules and the head of the epididymis. In this study, the objectives were: (1) to establish an in vitro system for the culture of epithelial cells from efferent ductules and initial segment epididymidis, (2) to validate the cultured epithelial cells using morphologic and functional characteristics, and (3) to test the hypothesis that estrogen mediates gene expression in cultured efferent ductule cells. The major findings are listed. Epithelial cells from efferent ductules and initial segment epididymidis can be maintained in the cultures. The cultured cells retained structural and epithelial characteristics, and maintained their in vivo capabilities of endocytosis and the expression of clusterin and estrogen receptor mRNAs. Thus, this in vitro system was suitable for the investigation of estrogen regulations in the efferent ductules and initial segment epididymidis. Differential display PCR analysis demonstrated that 17$\beta$-estradiol altered gene expression in cultured efferent ductule cells, and this effect was reversed by treatment with ICI 182,780, a pure antiestrogen. The differentially displayed products were tested in Northern blot hybridization and some transcripts were confirmed to be genes expressed in the efferent ductules under estrogen regulation. However, the nucleotide sequences of these genes did not match sequences of known genes from Genebank; therefore, these sequences were classified as novel, unidentified genes. Finally, clusterin, a marker for efferent ductule epithelial cells, was examined because it is not regulated by androgens. In this study, the expression of clusterin mRNA was significantly inhibited by the antiestrogen. In conclusion, estrogens regulate gene expression in the efferent ductules which is consistent with a recent discovery that disruption of estrogen receptor gene causes male infertility.
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