Cloning of estrogen-regulated messenger ribonucleic acids from rat uterus.

Abstract

A cDNA library prepared from the mRNA of uteri of estrogen-stimulated immature female rats was constructed in lambda gt10. Differential screening of the hybrid phages was performed using control and stimulated cDNAs as probes. Selected clones were then characterized by Northern and Southern blot hybridizations. In all, eight unique clones corresponding to estrogen-stimulated messages in rat uterus were identified. These clones hybridized to uterine mRNAs varying in size from 1.4-8.4 kilobases. Three of the clones were characterized as coding for three different types of collagen, and one as coding for smooth muscle actin. These are described in more detail elsewhere. The kinetics of increase in estrogen-regulated messages was examined. After a single injection of estradiol, five clones, including the three collagen mRNAs, showed two peaks of accumulation, at 4 and 24 h. Messages of two other clones were maximal at 12 h. The actin clone hybridized to mRNAs with peaks at 4 h for cytoskeletal actins and 8-12 h for smooth muscle actins. Sequential 24-h injections of the hormone produced multiple peaks of mRNA accumulation with a timing consistent with the kinetics found after a single injection of hormone. The multiple injections, however, did not result in enhanced mRNA accumulation for any of these clones. In fact, several messages showed suppressed accumulation with continued estradiol administration. Accumulated inhibitory factors in uterine cells may be responsible for this refractory condition. Except for the actin mRNA, the estradiol-stimulated mRNAs were expressed mainly in uterus and ovary. These clones may be useful in studies on the mechanism of action of estrogenic hormones and their tissue-specific regulation of gene expression.