LH/hCG-binding sites were measured in crude membrane fractions of porcine uteri. Specific high affinity and low capacity receptors for LH/hCG were found in all (n = 17) membrane preparations of myometrium but in only 5 of 17 crude membrane fractions of endometrium of porcine uteri. There was very little competition between hCG and porcine GH (pGH), bovine TSH, pFSH, and pPRL (0.5%, 0.3%, 0.2%, and less than 0.005%, respectively). Specificity of [125I]hCG binding to other tissues was determined by incubating crude membrane preparations of heart, skeletal muscle, liver, and kidney. Numbers and affinities of available LH/hCG-binding sites were characterized in all samples of myometrium and 5 endometrium membrane preparations that were positive for LH/hCG receptors. The results indicate that the number of uterine LH-binding sites in myometrium (0.66 +/- 0.17 fmol/mg) is 10 times less than the receptor capacity in porcine corpora lutea (7.46 +/- 0.54 fmol/mg) when expressed per mg protein of crude membrane preparation. However, it is approximately 60 times less when expressed per mg DNA equivalent of initial homogenate (1.31 +/- 0.28 vs. 81.18 +/- 3.64 fmol/mg, respectively). Receptor affinities of uterine LH/hCG-binding sites remained comparable to those of corpora lutea receptors (Ka = 7.8 X 10(10) M-1). Concentrations of LH/hCG-binding sites in myometrium taken from gilts in the late follicular phase of the estrous cycle (0.13 +/- 0.06 fmol/mg protein; n = 5) were significantly less (P less than 0.05 and P less than 0.01) compared to those in myometrium from luteal phase (0.85 +/- 0.22 fmol/mg protein; n = 6) or early pregnancy (1.03 +/- 0.15 fmol/mg protein; n = 6), respectively. This is probably the first evidence demonstrating specific binding of [125I]hCG by LH receptors in female uterine tissue.
The acute effects of oestradiol-17 p on the expression of the oestrogen receptor (ER) and insulinlike growth factor I (IGF-I) in the endometrium of ovariectomized pigs were examined. The steroid receptor level was assayed by hormone binding techniques and specific mRNAs analyzed by solution hybridization using %-labelled RNA probes complementary to the ligand-binding domain of the ER receptor gene and a 160 bp PanI-Pvul fragment of the IGF-I gene. One hour after a single injection of oestradiol (1 pg/kg BW), the nuclear oestrogen receptor (ER,) mean level was increased 3-fold whereas the ER mRNA content had not changed significantly. After 3 hours the ER, mean concentration was still high; the mean ER mRNA level had decreased by 15 % and the mean IGF-I mRNA had increased 3-fold above that in the samples collected prior to treatment from these ovariectomized animals. Six hours after the injection the ER, content had returned to the basal level and stayed there during the following six hours. The ER mRNA concentration continued to decline, reached its lowest value after six hours and had increased slightly by twelve hours. The IGF-I mRNA level increased steadily during the course of the experiment. At twelve hours after the injection it had increased 3-fold. From these data we conclude that in the pig uterus oestradiol down-regulates its own receptor and acts as a potent stimulator of endometrial growth by inducing IGF-I expression.
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