共查询到20条相似文献,搜索用时 15 毫秒
1.
L S Persianinov S V Sturchak E G Bobrova R N Shchedrina N D Fanchenko 《Biulleten' eksperimental'no? biologii i meditsiny》1975,79(5):93-96
It was demonstrated on the uteri of women and guinea pigs that estriol (in vitro) possessed a marked affinity to the estradiol-binding system of human and guinea pig uteri; the activity of steroid-receptor interaction of estriol in vitro constituted 9.4% for guinea pigs and 17% for man in relation to the estradiol activity. Administration of estriol to guinea pigs in vivo in a dose of 0.25-0.5 mg led to a sharp reduction of the estradiol-binding capacity of the receptor system of the uterus. It is supposed that there existed a competitive relationship between estradiol and estriol for binding with the active centres of the receptor proteins of the uterus. 相似文献
2.
M Tetsuo H Eriksson T Cronholm D Collins J Sj?vall 《Journal of steroid biochemistry》1989,33(3):371-378
The concentrations and turnover of estradiol isolated from cytosolic and nuclear fractions of uteri from ovariectomized rats given estradiol, either in single injections or in continuous infusion, were analyzed by gas chromatography-mass spectrometry. The analytical method was validated for different organs and lower limits of analysis were established. After infusion of 20 ng x h-1 for 18-22 h, mean estradiol levels were 2.0-2.4 fmol x mg-1 uterine wet weight in the nuclear fraction, and 1.2-1.5 fmol x mg-1 in the cytosolic fraction. The concentrations were about five times higher after a single injection of one microgram estradiol but the distribution between nuclear and cytosolic fractions was almost the same. The concentrations of estradiol in nuclei from liver and spleen were 50-200 times lower than those in uterus. Taken together with previous knowledge, the results indicate that the distributions of estradiol and its receptor are not the same and that hormone response cannot be predicted from the concentration of receptors alone. The exchange of estradiol molecules in the uterus was followed after a change of the infusion from unlabelled to [11,12,12-2H3]-labelled estradiol, or vice versa. The uterine uptake of estradiol was calculated to be about 0.7 fmol x h-1 x mg-1 uterine wet weight. The half-life time was calculated to be at least 4 h for estradiol molecules isolated from the nuclear fraction and 3 h (significantly shorter) for those isolated from the cytosolic fraction. The results indicate an uptake of 40-90% of all estradiol passing through the uterus in proestrus with only about 10% of available receptors becoming occupied. When the infusion was changed from estradiol to ethynylestradiol, estradiol disappeared from the uterus at the same rate as in the experiments above. Ethynylestradiol was taken up at a rate of about 0.3-0.4 fmol x h-1 x mg-1 tissue. The percentage of total steroid found in the nuclear fraction was higher for ethynylestradiol, about 70%, than for estradiol, about 60%, indicative of a more stable association of receptor to nuclear binding sites when ethynylestradiol is the ligand. 相似文献
3.
Estradiol-receptor complex from rat uterus has been shown to have an affinity for DNA-cellulose and ATP-Sepharose. This DNA and ATP binding of estradiol receptor was observed to be sensitive to low concentrations (0.01–0.2mM) of aurintricarboxylic acid. The inhibitor was more effective when added to preparations that contained activated estradiol-receptor complex. Steroid binding properties of the receptor remained intact under the above conditions as judged by charcoal adsorption assays and sucrose gradient analysis. In addition, a 40% inhibition in the nuclear translocation of cytosol estradiol receptor was observed when rat uteri were incubated with 10nM [3H] estradiol under an atmosphere of 95% O2 and 5% CO2 in the presence of aurintric-carboxylic acid. Our results suggest that aurintricarboxylic acid is an effective inhibitor of rat uterine estradiol receptor and that it may be acting by interfering with site(s) on the estradiol receptor which may be exposed upon activation and are subsequently involved in processes such as ATP binding, nuclear uptake and DNA binding. 相似文献
4.
1. The effect of a chronic morphine treatment on the in vitro contractile responses of the mouse uterus to adrenaline was studied. 2. Chronic morphine treatment induced a supersensitivity state in the uteri from both progesterone and estradiol treated mice. 3. The acute administration of morphine to the uteri from morphine tolerant-dependent and progesterone treated mice induced a further increase of the contractile effect of adrenaline. 4. Reserpine administration did not further increase the supersensitivity of the mouse uterus to adrenaline induced by a chronic morphine treatment. 5. Reserpine suppressed the acute effects of morphine in the uteri from tolerant-dependent mice. 相似文献
5.
《Journal of receptor and signal transduction research》2013,33(3):439-457
AbstractThe uterus of the guinea pig fetus has been shown to respond to estradiol treatment by an increase in uterine wet weight and a stimulation of the progesterone receptor protein. A study of the kinetics of these two parameters of estrogen response in the fetal uterus was undertaken in order to correlate these responses with changes in the estrogen receptor. Administration of estradiol to pregnant guinea pigs (1 mg/kg/body weight) leads to a rapid stimulation of the progesterone receptor by 6h after treatment which reaches maximal values by 15.5h, which are increased 7-fold in estradiol-primed guinea pigs above values in untreated animals. The estradiol receptor undergoes rapid translocation from the cytosol into the nucleus by 1h after hormone treatment and is retained in the nucleus for at least 6h. At the same time, there is a 50% decrease in the total occupied and available estradiol receptor concentration at 6h after treatment. Estradiol treatment also provokes an increase in wet weight of the fetal uterus which is significantly greater after 3 consecutive days of treatment (171% ± 24 (S.D.) above wet weights of untreated uteri which were considered as 100%) than after only 1 day (121% ± 25 (S.D.)). These estrogen responses were found to be of long duration since uterine wet weights and progesterone receptor concentrations remained well above control values even 5 days after a single treatment with estradiol. In conclusion, the fetal uterus responds to estradiol treatment by a slow increase in wet weight and a rapid stimulation of the progesterone receptor protein with a concomitant loss in estradiol receptor concentration. 相似文献
6.
Chronic estrogen treatment causes an alteration in uterine estrogen receptor dynamics of rats 总被引:1,自引:0,他引:1
B Sato Y Nishizawa K Noma S Kishimoto K Matsumoto 《Biochimica et biophysica acta》1983,755(3):412-419
Estrogen receptor content and dynamics in the uteri obtained from chronically estrogenized rats were analyzed. 12 day treatment with a subcutaneous implantation of a diethylstilbestrol pellet resulted in maximal stimulation of uteri with regard to wet tissue weight, DNA content, as well as progesterone receptor content without significant alteration of the estrogen receptor level. Estrogen receptor dynamics in just ovariectomized or ovariectomized and diethylstilbestrol-stimulated rats elicited by a single injection of estradiol were next examined using the exchange methods. The cytosol receptor content rapidly declined, with a small and temporary accumulation of the nuclear receptor in the uterus from rats continuously exposed to diethylstilbestrol during the preceding 12 days. A relatively rapid cytosol receptor replenishment was also observed in rats pretreated with diethylstilbestrol. This was accompanied by a rapid decrease in the nuclear receptor level to 70% of the preinjection value at 5 h after estradiol administration. These data are in contrast to findings on uteri of ovariectomized and nonestrogen-treated rats, in which a single injection of estradiol resulted in a prolonged nuclear receptor retention and a delayed cytosol receptor replenishment. Adrenalectomy did not result in a significant change of receptor dynamic patterns, suggesting that adrenal steroids do not play a role in the alteration of receptor dynamics elicited by continuous stimulation with diethylstilbestrol. These observations suggest that a continuous exposure of rat uteri to the estrogen causes an altered regulation of estrogen receptor dynamics by the homologous steroid compared to those in chronically estrogen-deprived rats. 相似文献
7.
The present experiments report the effects of estradiol or of progesterone on the activity of 15-prostaglandin-dehydrogenase (PGDH) in the uterus of spayed rats. When the substrate was PGF2 alpha the treatment with progesterone (4 mg X day-1, two days) or with estradiol-17-beta (0.5 ug + 1 ug) did not show any effect on the activity of the enzyme. On the contrary, uteri from ovariectomized rats injected with a higher dose of estradiol-17-beta (0.5 ug + 50 ug) exhibited a significant increment. When the substrate was PGE2, progesterone failed again to modify the enzyme activity, whereas estradiol, both at a low and at a high doses, enhanced significantly the uterine PGDH activity. The possibility of two different PGDHs for each PG and the role of estradiol in enhancing PGE2 catabolism into 15-keto-PGE2 as a mechanism subserving the effect of estrogens on the output of this PG in the rat uterus, are discussed. 相似文献
8.
Estradiol receptor: phosphorylation on tyrosine in uterus and interaction with anti-phosphotyrosine antibody. 总被引:3,自引:1,他引:2
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Estradiol receptor from rat uteri incubated with [32P] orthophosphate has been purified by diethylstilbestrol--Sepharose followed by heparin--Sepharose chromatography. The purified receptor, analyzed by centrifugation through sucrose gradients after incubation with monoclonal antibodies against purified estradiol receptor, appears to be labeled with 32P. The receptor preparation has been further purified by immunoaffinity chromatography and submitted to SDS--poly-acrylamide gel electrophoresis. A heavily 32P-labeled 68 kd protein and a very lightly 32P-labeled 48 kd protein, probably a proteolytic product of the 68 kd protein, were detected. Phosphoamino acid analysis of the receptor eluted from the immunoaffinity column shows that its 32P-labeling occurs exclusively on tyrosine. This is the first report on phosphorylation on tyrosine of a steroid receptor in tissue. It is consistent with our previous finding that a uterus estradiol receptor-kinase, which confers hormone binding ability to the estradiol receptor, in vitro phosphorylates this receptor exclusively on tyrosine. Calf uterus receptor binds with high specificity and affinity to monoclonal anti-phosphotyrosine antibodies covalently bound to Sepharose (Kd = 0.28 nM). Dephosphorylation of the receptor by nuclei containing the calf uterus nuclear phosphatase abolishes the interaction with antibodies. These results suggest that also in calf uterus, estradiol receptor is phosphorylated on tyrosine. Anti-phosphotyrosine antibodies bound to Sepharose have been used to partially purify the estradiol receptor from calf uterus. 相似文献
9.
The naturally occurring C-17 fatty acid esters of estradiol are long-acting estrogens 总被引:4,自引:0,他引:4
C-17 fatty acid esters of estradiol are naturally occurring biosynthetic metabolites of estradiol. A representative component of this family of esters, estradiol-17-stearate, was studied in order to determine the estrogenic properties of these unusual hydrophobic steroids. Following the classical estrogen bioassay, a solution of this ester in oil was injected subcutaneously into immature rats once a day for 3 days. There was little effect on the uterus on the first day after the third injection. However, on subsequent days a large stimulation of uterine growth occurred. The course of this estrogenic effect was exactly opposite to that obtained with estradiol. In order to eliminate the possibility that this effect on the time course of estrogenic stimulation was caused by increased solubility of the hydrophobic esters in the carrier oil, the steroids were administered to adult ovariectomized animals in aqueous medium via a single intravenous injection. The uterotrophic response to estradiol was maximal at 12 h and was completely dissipated in 48-60 h. Estradiol-17-stearate produced a uterotrophic effect of twice the duration of estradiol. In the immature rat, aqueous intravenous injections of estradiol-17-stearate produced a greater uterotrophic effect than estradiol and this effect was still maximal 96 h later. In addition, this single injection of estradiol-17-stearate advanced the time of vaginal opening, a marker for puberty in the female rat. The mechanism of the prolonged estrogenic stimulation was investigated by studying the steroidal content of the uterus after injecting [3H]estradiol and [3H]estradiol-17 -stearate i.v. into immature rats. At 1 and 4 h there was significantly more radioactivity in the uteri of the [3H]estradiol treated animals. At later times (8 h and onwards) the total radioactivity in the uterus did not differ appreciably between the two groups. However at these later times, the amount of [3H]estradiol was far greater in the uteri of animals receiving [3H]estradiol-17-stearate. Consequently, the prolonged estrogenic effects of the endogenous C-17 fatty acid esters of estradiol are caused by the increased duration of the estrogenic signal. It is hypothesized that one of the roles of the fatty acid is to protect the steroid nucleus from metabolism and thereby prolong the life of the parent C18 steroid. Thus, the results of these experiments are consistent with the family of endogenous alkyl esters of estradiol having a physiological role as long-acting estrogens. 相似文献
10.
Physiological evidence is presented for the possible existence of anhydrolevuglandin E2-like activity in extracts of uteri from diestrous rats and after treatment of adult rats with estradiol and progesterone. The extracts were able to inhibit contractions in rat uterine preparations stimulated by PGE2. Uteri of vaginal Stage 3 (metestrus) were quiescent and showed decreased responsiveness to PGE2 and PGF2 alpha. These uteri showed some contractility when incubation medium from diestrous uteri (Stage 5) were transferred to them and incubation medium from them inhibited the contractility of Stage 5 uteri. When incubation media were exchanged between contractile uteri from of stages other than Stage 3, there was no change in the contraction patterns. Taken together, we believe these data indicate that AnLGE2 may be a normal constituent of the rat uterus and is physiologically increased during Stage 3 (metestrus) of the estrous cycle. 相似文献
11.
N. Sterin-Speziale M.F. Gimeno A. Bonacossa A.L. Gimeno 《Prostaglandins & other lipid mediators》1980,20(2):233-243
The motility of isolated uterine horns as well as the generation of PGE and PGF like material by the uterus from estrus and spayed rats, treated or untreated with 17-beta estradicl, were studied. Following 40 minutes of mounting the spontaneous motility of uteri from estrus rats had a lower magnitude than that from spayed ones. The amount of PGF-like material was similar in both groups whereas the first one liberated less PGE-like substance. In spayed animals treated with 1 μg of 17-beta estradiol the decay of spontaneous contractile force was higher than that observed in untreated rats, and similar to that displayed by uteri from estrus. Less PGE-like material was liberated in comparison with spayed animals and a tendency to produce higher quantity of PGF-like compounds was observed, although the level was not significantly different. With 50 μg of 17-beta estradiol the spontaneous reduction of contractile activity was higher than in spayed animals and than in those treated with 1 μg. The amount of PGF-like material liberated was higher than in spayed rats and less PGE-like substance was generated comparing with spayed and 1 μg-treated animals. These findings show that estradiol decreases the release of PGE-like compound. It would also appear that this may have some relationship with the levels of spontaneous contractile activity of the isolated rat uterus. 相似文献
12.
Expression and activation of Akt/protein kinase B in sexually immature and mature rat uterus 总被引:1,自引:0,他引:1
Lengyel F Vértes Z Kovács KA Környei JL Sumegi B Vértes M 《The Journal of steroid biochemistry and molecular biology》2004,91(4-5):285-288
This study investigated the expression and activation of Akt/PKB in developing and adult rat uterus. Expression of Akt was observed in uteri from adult ovariectomized and 7–35-day-old rats and no changes were observed in response to in vivo estradiol treatment (1–100 μg/100 g b.w.). To examine the mechanisms of PKB/Akt activation, phosphorylation at Thr308 and Ser473 regulatory sites were studied in uteri. Akt was constitutively phosphorylated on Ser473 residue in the untreated, control uteri, while phosphorylation of Thr308 was observed only after estradiol 17β (E2) treatment. The effects of E2 treatment were age dependent, no response was induced in 11-day-old uteri, while in 28 days and older rats the activation of Akt at both regulatory sites, Ser473 and Thr308, increased, the first response was detected 2 h after treatment, reaching the highest rate at 6 h. The rate of phosphorylation was stronger at Ser473 residue. The results suggest that the regulation of Akt activation at two regulatory sites in rat uteri are different, phosphorylation of Thr308 seems to be entirely estrogen dependent, while the phosphorylation of Ser473 is regulated by other factors as well as estrogen. 相似文献
13.
A.M. Franchi A. Faletti M.F. Gimeno A.L. Gimeno 《Prostaglandins & other lipid mediators》1985,29(6):953-960
The present experiments report the effects of estradiol or of progesterone on the activity of 15-prostaglandin-dehydrogenase (PGDH) in the uterus of spayed rats. When the substrate was PGF2α the treatment with progesterone (4 mg.day−1, two days) or with estradiol-17-beta (0.5 ug + 1 ug) did not show any effect on the activity of the enzyme. On the contrary, uteri from ovariectomized rats injected with a higher dose of estradiol- 17-beta (0.5 ug + 50 ug) exhibited a significant increment. When the substrate was PGE2, progesterone failed again to modify the enzyme activity, whereas estradiol, both at a low and at a high doses, enhanced significantly the uterine PGDH activity. The possibility of two different PGDHs for each PG and the role of estradiol in enhancing PGE2 catabolism into 15-keto- PGE2 as a mechanism subserving the effect of estrogens on the output of this PG in the rat uterus, are discussed. 相似文献
14.
《Journal of receptor and signal transduction research》2013,33(3):415-438
AbstractThe effects of salt-extraction on type I and type II estrogen binding sites were examined in uterine nuclei. Injection (10 ug) of estradiol or estriol in adult ovariectomized rats induced maximum numbers (80–100%, ~ 1 pmole/uterus) of 0.4 M KCL resistant type I estrogen complexes at 1 hour. Only estradiol, which sustained these levels for long periods of time (4–24 hours) stimulated true uterine growth.Likewise, a single injection of estradiol, but not estriol, also elevated nuclear type II sites throughout the entire uterine growth period (1–48 hours). However extraction of these nuclei from estradiol injected rats with 0.4 M KCL increased the numbers of type II sites from ~ 1 pmole/uterus (non-extracted nuclei) to ~ 8 pmoles/uterus (salt resistant plus salt-extractable fractions). Sixty percent of these sites were resistant to salt-extraction. Continuous exposure to either estradiol or estriol by beeswax implants stimulated nuclear type II sites which were highly resistant (80%) to KCL-extraction, and additional sites were not exposed by high salt. Thus chronic treatment with both estrogens “locked in” nuclear type II sites such that they were resistant to KCL-extraction. This resistance of type II sites to salt-extraction correlated with the ability of estradiol and estriol implants to stimulate true uterine growth. The procedures presented here for nuclear preparation and assay have reduced non-specific binding considerably in the uterine system, and may eliminate the need to perform exchange assays on salt-extracted nuclei in other systems. 相似文献
15.
Comparison of media proteins from ovariectomized rat uteri following estrogen treatment 总被引:1,自引:0,他引:1
Ovariectomized rats were treated with estradiol for 3 days after which their uteri were incubated in vitro and radioactive media proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Media were also chromatographed on G-25 Sephadex Blue Sepharose columns to isolate subsets of proteins. The results demonstrate that two proteins are consistently increased following estrogen treatment. These proteins have molecular weights of 104,000 and 65,000. Neither protein binds to Blue Sepharose to a great extent. The use of the protein synthesis inhibitors, emetine and actinomycin D, demonstrates that the proteins are synthesized de novo. These two proteins may serve as markers for genomic response to estradiol in the rat uterus. 相似文献
16.
Summary The localization and retention of radioactivity in the uteri of fetal and newborn guinea pigs was studied after subcutaneous injection of 3H-progesterone into intact and estradiol-primed animals. In the fetal uterus of the animals treated with estradiol, a significant increase in the accumulation of silver grains was observed, mainly localized in the stroma and myometrium, but very little is detected in the uterine gland. On the other hand, in the uteri of newborns, the effect of estradiol on radioactivity retention was significantly less intense and the radioactivity is mainly localized in the epithelium and the uterine gland. These data correlate well with the quantitative evaluation of the concentration of progesterone-specific-binding sites in the non-treated and in the estradiol-primed fetal and newborn guinea pigs. 相似文献
17.
T Toko K Matsuo J Shibata K Wierzba M Nukatsuka S Takeda Y Yamada T Asao T Hirose B Sato 《The Journal of steroid biochemistry and molecular biology》1992,43(6):507-514
DP-TAT-59, (Z)-2-(4-(1-(4-hydroxyphenyl)-2-(4-isopropylphenyl)-1-butenyl) phenoxy)-N, N-dimethylethylamine, has been reported to inhibit estrogen-stimulated growth of MCF-7 cells as well as rat uterus at lower concentrations than the hydroxymetabolite of tamoxifen (4-OH-TAM). In the present study, the growth of mouse Leydig cell tumor, B-1F cells were also more effectively inhibited by DP-TAT-59 than 4-OH-TAM. Additionally, the expression of estrogen responsive element ligated CAT gene transfected into B-1F cells was also suppressed by DP-TAT-59. Thus, the interaction of DP-TAT-59 with estrogen receptor (ER) was characterized and compared with that of 4-OH-TAM using immature rat and bovine uteri. The dissociation constant of DP-TAT-59 to ER of immature rat uterus was 0.24 nM and was similar to that of 4-OH-TAM (Kd = 0.20 nM) and estradiol (Kd = 0.29 nM). Using sucrose density gradients, the sedimentation constant of DP-TAT-59 with bovine uterus was 4.9S, which was similar to that of estradiol (5.1S) and 4-OH-TAM (5.3S). However, the elution profile of the DP-TAT-59-ER complex from a DEAE-Sephadex column was different for both estradiol-and 4-OH-TAM-ER complexes. These results suggest that ER forms different complexes with DP-TAT-59 than estradiol or 4-OH-TAM, while the ER binding affinity of these compounds are similar to each other. 相似文献
18.
The normal time for cleavage to the 2- and 8-cell stages of development of naturally ovulated hamster embryos is 25–27.5 and 59–61 h respectively, after ovulation and mating. The corresponding values for PMS-hCG treated hamsters were 33–35.5 and 62–64 h respectively. When 2-cell embryos, obtained by this timing method, were transferred to the oviductal bursa on Day 2 of the cycle, and when 8-cell embryos were transferred to the uterus on Day 3 of the cycle, implantation rates of 61.5 and 64.0% respectively, were obtained. The current study was conducted to determine the effects of transfer on estradiol and progesterone uptake by the uterine tissue on Day 14 after mating. A 2-fold increase in estradiol uptake was observed in superovulated, nonpregnant uteri when compared with nonsuperovulated animals. This level was also significantly higher than with nonsuperovulated pregnant animals and of animals receiving 2-cell embryo transfers. Estradiol uptake increases of 3.2 and 1.2-fold were noted for animals receiving 8-cell embryo transfers in naturally and superovulated groups respectively. Superovulation resulted in increased progesterone uptake. Transfer of 2- and 8-cell embryos resulted in a 15.8 and 109.9% increase in progesterone uptake respectively in naturally ovulated hamsters. Similar values for superovulated hamsters were 16.2 and 87.6% respectively. The 8-cell embryos, however, were transferred about four hrs prior to the time for normal 8-cell cleavage and this, coupled with increased estradiol uptake by the embryos themselves, resulted in an elevated estradiol and progesterone uptake in the uterine tissue even when measured on Day 14 of pregnancy. The degree of increase was less with superovulated animals receiving 8-cell embryos, reflecting higher levels of estradiol and progesterone uptake in control tissues. This could account for the slight delay in development of superovulated embryos. 相似文献
19.
Estradiol binding components in the cytosol and nuclear fractions of the ovary from immature rats (22–28 days old) were characterized by methods. Several of the biochemical characteristics of the estradiol binding components in the ovarian tissue were compared with the estradiol receptor from the uterus. The results suggest that the ovarian estradiol binding components are similar to the specific high affinity estradiol receptors in the uterus. In the cytosol of intact rat ovary a significant fraction of the total binding sites was found to be occupied, presumably by the endogenous estrogen. Following hypophysectomy there was a significant increase in the available cytosol binding sites. Evidence for translocation of cytosol receptor-estrogen (RE) complex to the nucleus was obtained for the ovary. The sedimentation properties of the RE complex of the ovary and the uterus are similar. The ovarian cytosol RE complex sediments at 7-8S in glycerol gradients at low ionic strength and at 4S in sucrose gradients at high ionic strength. Following extraction with 0.4 M KCl the ovarain nuclear RE complex sediments at 5S in sucrose gradients which is identical to that of the uterine nuclear receptor. 相似文献
20.
Estrogen regulation of tissue-specific expression of complement C3 总被引:14,自引:0,他引:14
S A Sundstrom B S Komm H Ponce-de-Leon Z Yi C Teuscher C R Lyttle 《The Journal of biological chemistry》1989,264(28):16941-16947
The administration of estradiol to immature rats results in the increased synthesis and secretion of a 180-kDa protein, composed of 115- and 65-kDa subunits, by the uterine luminal epithelial cells. A monoclonal antibody against the 180-kDa protein was utilized to isolate the corresponding cDNA (LE-1) from a rat uterine luminal epithelial cell cDNA lambda gt11 expression library. This LE-1 cDNA was sequenced and shown to be homologous to complement component C3. The sequence was approximately 81 and 90% homologous to human and mouse C3, respectively. The LE-1 cDNA sequence was homologous with the 3' portion of the C3 mRNA containing the alpha subunit (115 kDa). Uterine mRNA isolated from immature rats treated with 1 microgram of estradiol for 24 h demonstrated a 25-fold increase in the concentration of a 6.0-kilobase mRNA by Northern hybridization with either LE-1 or authentic human C3 cDNA probes. To further examine the possibility that the estradiol-regulated secretory protein was C3, an aliquot of radiolabeled media protein from control and estradiol-stimulated rat uteri was incubated with goat anti-rat C3 antibody. The immunoprecipitated radiolabeled protein from estradiol-treated animals was increased significantly (p less than 0.01) compared to media from control animals. Analysis of the immunoprecipitated proteins on nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a single protein of 180 kDa from estradiol-stimulated uterine media, whereas no detectable proteins were immunoprecipitated from media obtained from control uteri. Also, when the immunoprecipitated protein was reduced (20 mM dithiothreitol) it dissociated into two subunits of 115 and 65 kDa. Immunohistochemical studies demonstrated the presence of C3 only in the epithelial cells of estrogen-stimulated rat uteri. In addition, the estradiol-stimulated mRNA was only detectable in uterine epithelial cell RNA. Analysis of liver RNA demonstrated a 6.0-kilobase mRNA, as in the uterus, when hybridized with LE-1. However, unlike the uterus, its concentration was not influenced by estrogen administration with up to three daily injections of 100 micrograms of diethylstilbestrol. Based on biophysical, DNA sequence, and antibody data we conclude that rat uterine epithelial cells produce C3 in response to estradiol whereas the expression in the liver was not modulated by estrogens. 相似文献