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Endocrinology Vol. 138, No. 11 4613-4621
Copyright © 1997 by The Endocrine Society

ARTICLES

Tissue Distribution and Quantitative Analysis of Estrogen Receptor-{alpha} (ER{alpha}) and Estrogen Receptor-ß (ERß) Messenger Ribonucleic Acid in the Wild-Type and ER{alpha}-Knockout Mouse

John F. Couse, Jonathan Lindzey, Kaj Grandien, Jan-Åke Gustafsson and Kenneth S. Korach

Receptor Biology Section (J.F.C., J.L., K.S.K.), Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709; and the Department of Medical Nutrition (K.G., J.-A.G.), Karolinska Institute, NOVUM, S-14186, Huddinge, Sweden

Address all correspondence and requests for reprints to: Dr. Kenneth S. Korach, Receptor Biology Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, MD B3–02, P.O. Box 12233, Research Triangle Park, North Carolina 27709.

Until recently, only a single type of estrogen receptor (ER) was thought to exist and mediate the genomic effects of the hormone 17ß-estradiol in mammalian tissues. However, the cloning of a gene encoding a second type of ER, termed ERß, from the mouse, rat, and human has prompted a reevaluation of the estrogen signaling system. Based on in vitro studies, the ERß protein binds estradiol with an affinity similar to that of the classical ER (now referred to as ER{alpha}) and is able to mediate the effects of estradiol in transfected mammalian cell lines. Essential to further investigations of the possible physiological roles of ERß, and its possible interactions with ER{alpha}, are data on the tissue distribution of the two ER types. Herein, we have described the optimization and use of an RNase protection assay able to detect and distinguish messenger RNA (mRNA) transcripts from both the ER{alpha} and ERß genes in the mouse. Because this assay is directly quantitative, a comparison of the levels of expression within various tissues was possible. In addition, the effect of disruption of the ER{alpha} gene on the expression of the ERß gene was also investigated using the ER{alpha}-knockout (ERKO) mouse. Transcripts encoding ER{alpha} were detected in all the wild-type tissues assayed from both sexes. In the female reproductive tract, the highest expression of ERß mRNA was observed in the ovary and showed great variation among individual animals; detectable levels were observed in the uterus and oviduct, whereas mammary tissue was negative. In the male reproductive tract, significant expression of ERß was seen in the prostate and epididymis, whereas the testes were negative. In other tissues of both sexes, the hypothalamus and lung were clearly positive for both ER{alpha} and ERß mRNA. The ERKO mice demonstrated slightly reduced levels of ERß mRNA in the ovary, prostate, and epididymis. These data, in combination with the several described phenotypes in both sexes of the ERKO mouse, suggest that the biological functions of the ERß protein may be dependent on the presence of ER{alpha} in certain cell types and tissues. Further characterization of the physiological phenotypes in the ERKO mice may elucidate possible ERß specific actions.




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