Further Studies on Rna and Protein Biosynthesis in the Ovariectomized Mouse Uterus

Abstract

1. RNA was isolated from the ovarian and uterine tissues of calves by the cold phenol procedure. 10 OD of uterine-RNA, ovarian-RNA or the equivalent volume of normal saline was injected into the uterine lumen of ovariectomized mice.2. Alkaline phosphatase and \g=b\-glucuronidase levels in the uterus were studied 3 days following the intra-uterine injections.3. Results indicate that both U-RNA and O-RNA were able to induce protein (enzyme) production in the atrophied mouse uterus. Although U-RNA caused higher \g=b\-glucuronidase content of the uterus than O-RNA, the physiological state of the tissue used to supply the RNA did not have any effect. 4. Ribonuclease and KOH digestion of the RNA extract abolishes its action. Actinomycin D given simultaneously with RNA represses the latter's action.5. The possibility of hormonal contamination in the RNA extraction is discussed and should be ruled out after ether washing.Recent studies support the concept that steroid hormones act on nuclear DNA to stimulate messenger-RNA (m-RNA) synthesis (Gorski et al. 1965;Kochakian 1962). This m-RNA initiates biochemical and structural changes in the uterine cells following hormonal application (Ui & Mueller 1963). Such interpretation is substantiated by various studies on protein biosynthesis in vivo (Coster Sc Kochakian 1962; Williams-Ashman et al. 1963) and in vitro {Wilson 1962), nucleic acid fluctuation in castrate and hormone treated animals (Wicks et al. 1965; Kochakian 8c Harrison 1962), local puromycin application to oestrogen treated rats (Talwar Sc Segal 1963) and in situ injections of RNA in the uterine

The Effect of Exogenous Ribonucleic Acid on Alkaline Phosphatase Activity in the Ovariectomized Mouse Uterus

“…Our data on oestradiol-3H showed that thorough washing of RNA with ether removes all traces of the hormone. This confirms our previously reported contention (Mansour 1967), along with similar findings of Segal 10£-A B-RNA 10•--• saline injected controls 10et al (1965), that hormonal contamination is not a factor in these studies pro¬ vided that the RNA is washed with ether.…”

“…Liver-RNA (L-RNA) injected into the uterus of ovariectomized mice (Man¬ sour 8c Niu 1965; Mansour 1967) and rats (Segal et al 1965) failed to give the biochemical and structural changes observed after uterine-RNA injections. On the other hand, L-RNA extracted from tissues preincubated with 17/5-oestradiol without ether washing of the RNA, caused a significant increase of the alkaline phosphatase content of the atrophied mouse uterus.…”

“…Previous work has shown that RNA from an appropriate source had the capacity to stimulate alkaline phosphatase, in uteri deficient in this enzyme (Mansour 8c Niu 1965;Mansour 1967). Hormonal contamination of the in¬ jected RNA was excluded on the assumption that steroids are extractable with alcohol and ether, both of which were used in the RNA extraction.…”

Hormonal Regulation of the Reproductive Tract in Female Mammals

“…Actinomycin D treatment does not block the cellular hypertrophy induced by active RNA extracts (169 (116) demonstrated that when RNA prepared from the uteri of mature cycling mice was administered by intrauterine injection to spayed mice, the biosyn thesis of alkaline phosphatase was increased and the normal adult appearance of the endometrium was restored. The ability of RNAs from bovine ovary and uterus to stimulate the local response and to increase mouse uterine ,a-glucuronidase activity was reported in a later study (114). The possibility that contaminating estradiol is the basis for the RNA effect was considerably lessened when RNA prepared fr om bacteria, which possess an alkaline phosphatase and no known estrogenic activity, and injected into the ovariectomized mouse uterus stimulated alkaline phosphatase activity (liS).…”

Decidual Alkaline Phosphatase Activity in the Pregnant and Pseudopregnant Rat

“…The effects of estrogens on uterine ALP in ovariectomized mice or rats may be inhibited by administration of actinomycin D21, 27,28 or estrogen antagonists.29,:jo The increase in the endometrial enzyme may be induced by instilling RNA extracted from uteri of estrogen-treated mice into the lumina of uteri of ovariectomized mice. 31,32 These findings suggest that, in the mouse, estrogen stimulates DNA-directed synthesis of RNA which, in turn, is responsible for the synthesis of enzyme protein. 21,27,28.31.32 Unlike the findings in the above mentioned species, the endometrial ALP activity in the cow33,34 and hamster3j appears to be stimulated by progesterone rather than by estrogens.…”