Estradiol induction of accelerated energy metabolism in prepuberal rat uteri in vitro: mRNA hybridization and [13C]NMR studies.

In vitro treatment with 30 nM 17 beta-estradiol stimulated the induction of mRNA for the brain type isozyme of creatine kinase BB (CKBB) and stimulated glucose metabolism in perifused uteri from 27-29-day-old rats. The perifusion conditions maintained the normal NMR spectrum of high energy phosphates for at least 24 h. This technique permitted the demonstration that perifused rat uteri stimulated by 17 beta-estradiol show increased mRNA for creatine kinase BB, 1 h after estrogen addition. The time-course of increase, measured by Northern blot hybridization, parallels that seen in mRNA extracted from uteri after in vivo induction by i.p. injection of 5 micrograms 17 beta-estradiol; the maximal increase is seen at 2-4 h. Experiments utilizing actinomycin D (4 micrograms/ml) for inhibition of RNA synthesis showed that CKB mRNA from both untreated and estradiol stimulated uteri had a similar half-life, of approximately 2 h, indicating that CKB mRNA is transcriptionally regulated. In the same system, the rate of glycolysis was measured by NMR spectroscopy using [1-13C]glucose. Following in vitro stimulation with 30 nM estradiol, glycolysis increased within 3 h, in parallel to increases previously found in uteri from in vivo stimulated rats.