Estrogen Receptor mRNA Levels Research Articles

Altered Placental mRna Expression of the Progesterone but not the Estrogen Receptor in Pregnancies Complicated by Preeclampsia

Objective: To determine whether the mRNA expression of the progesterone receptor (PR) or the estrogen receptor (ER) was altered in pregnancies complicated with preeclampsia (PE), small for gestational age (SGA), or PE with SGA. It has been indicated that not only estrogen but also progesterone may influence the vascular system during pregnancy, although the mechanism remainns unclear. It has also been shown that the placentas from women with PE produce more progesterone than those from normal pregnancies.Methods: Tissue samples from the central part of fetal placenta were obtained immediately after delivery from 20 women with normal pregnancies, 11 women with PE, 8 women with SGA, and 6 women with both PE and SGA.Main Outcome Measures: Total nucleic acids were prepared and mRNA levels of PR and ER were measured by a solution hybridization technique.Results: The mRNA levels of ER did not differ among the groups studied. The mRNA expression of PR was significantly higher in the placentas with PE both with and without SGA than in normal placentas. However, there was no change in the mRNA level of PR in the group with SGA as a single entity.Conclusion: The observation of an altered mRNA level of PR in PE with or without SGA could shed new light into regulation of the uteroplacental blood flow in this disease.

Expression of the Estrogen and Progesterone Receptors and Insulin-like Growth Factor-I Genes in the Porcine Myometrium during Early Pregnancy.

After conception and during early pregnancy in pigs, uterine motility plays a keyrole in sperm transport and embryo spacing prior to placentation. This process is modulated by gonadal steroids. Early pig embryos secrete estradiol with a peak d. 12-13 and estradiol elicits part of its influence on target cells in the genital tract via the insulin-like growth factor-I (IGF-I). Therefore, in the present study, the levels of the estrogen receptor (ER) and progesterone receptor (PR), as well as their mRNAs and the IGF-I mRNA, were measured in the myometrium of early pregnant pigs (days 8-30 after onset of estrus) by EIA and solution hybridization, in order to establish relationships among the expressions of IGF-I, ER and PR during early pregnancy. The ER level was low on days 8-10 and decreased even further to very low levels on days 25-30 while the level of ER mRNA was unchanged during the same period. The PR level was decreasing during the days monitored, although PR mRNA showed a tendency to increase from days 12-13 to days 25-30. The level of myometrial IGF-I mRNA was higher on days 25-30 as compared to days 8-18 indicating that myometrial IGF-I mRNA expression is not affected by the production of conceptus estradiol on days 12-13. Significant correlations were found between the expressions of IGF-I mRNA and PR mRNA and between the levels of ER and PR. The results suggest that in the porcine myometrium the expressions of IGF-I and PR genes, respectively ER and PR, may have common regulating factors.

Regulation of estrogen receptor mRNA by 2,3,7,8-tetrachlorodibenzo-p-dioxin as measured by competitive RT-PCR.

Environmental contaminants, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), cause alterations in gene expression. In this study, we measured the regulation of estrogen receptor (ER) mRNA in female CD-1 mice by competitive RT-PCR. Previous work suggests that ER protein levels are affected by TCDD, but how this is regulated is uncertain. These studies found no significant changes in ER mRNA levels, but the methods used (Northern blot analysis and RNase protection assays) lack sensitivity for measuring the low levels of RNA transcript, such as ER mRNA. The method described here offers an excellent alternative for quantifying the changes in mRNA levels. Internal competitors were created with gene-specific primers for ER and beta-actin by PCR reactions at low annealing temperatures. For each sample, the mRNA levels of ER and beta-actin were determined. Using competitive RT-PCR, the relative changes in ER mRNA from TCDD-treated and control animals were determined after normalization with the levels of beta-actin mRNA. The ER mRNA from female CD-1 mice treated with TCDD (single dose 5 micrograms/kg, i.p., 4 days) was found to be significantly suppressed as compared with the vehicle control in all tissues examined. TCDD decreased ER mRNA in the liver (30.1%) as expected. However, the greatest effect was in the reproductive tissues, with a 64.2% reduction in ER mRNA in the ovary. This is the first demonstration that TCDD causes tissue-specific downregulation of ER mRNA. These effects may contribute to the tissue-specific toxicity of TCDD.

Hormonal regulation of the estrogen receptor in primary cultures of hepatocytes from female rats

Estrogen treatment affects the hepatic synthesis and/or secretion of several proteins involved in clinically important pathological processes such as atherosclerosis, hypertension, and thrombosis. The endocrine regulation of the estrogen receptor (ER) concentration in primary cultures of rat hepatocytes was studied. Human growth hormone (hGH) and dexamethasone (DEX) in combination increased ER concentration 6-fold and ER mRNA levels 2.5-fold. These effects were not significantly different from those observed after treatment with the purely somatogenic bovine growth hormone (GH) in combination with DEX. Treatment with the lactogen ovine prolactin in the presence or absence of DEX did not significantly affect ER or ER mRNA concentrations. Triiodothyronine treatment at the most effective concentration (50 n m) increased ER and ER mRNA levels twofold. Medium supplementation with estradiol (0.1 n m) throughout the experiment did not affect the response to treatment with hGH and DEX. Treatment with high concentrations of ethinylestradiol in combination with hGH and DEX, however, increased the ER level twice as much as hGH and DEX without addition of estradiol or ethinylestradiol, whereas the ER mRNA concentration was the same in both the GH + DEX group and GH − DEX + (estradiol or ethinylestradiol) groups. These data indicate the importance of GH in combination with glucocorticoids for the maintenance of ER concentrations in the rat liver. Thyroid hormones may be of some, although minor importance, whereas the data suggest that prolactin is not directly involved in hepatic ER regulation

Enhancement of estrogen receptor gene expression in the mediobasal hypothalamus during anestrus in the beagle bitch

Estrogen receptor mRNA (ER mRNA) levels were measured in the mediobasal hypothalamus (MBH) of beagle bitches at different stages of the estrous cycle, and compared with levels in ovariectomized (OVX) estrogen-treated bitches. In cyclic bitches, the level of hypothalamic ER mRNA increased during the progression of anestrus and declined thereafter. Hypothalamic ER mRNA and plasma luteinizing hormone (LH) levels during anestrus and proestrus were positively correlated ( r=0.94, P<0.001). In OVX bitches, levels of hypothalamic ER mRNA were low, and increased significantly after treatment with a low dose of estradiol benzoate. These results suggest that, during the course of anestrus in the bitch, hypothalamic ER mRNA expression increases, and may be up-regulated by estradiol.

Estradiol up-regulates estrogen receptor and progesterone receptor gene expression in specific ovine uterine cells.

The regulation of estrogen receptor (ER) and progesterone receptor (PR) genes is critical to estrogen and progesterone responsiveness of the uterus during the estrous cycle. A low dose of estradiol, given to ovariectomized ewes to mimic the preovulatory estrogen surge, acutely enhanced ER and PR gene expression in most uterine cells. Estradiol effects were measured at 12, 24, and 48 h post-injection (n = 6 ewes per time) with immunohistochemistry and in situ hybridization. Whereas vehicle-treated ovariectomized ewes demonstrated low to moderate ER and PR mRNA and protein expression, estradiol enhanced PR mRNA and protein expression (at 12 h and 24 h, respectively) more rapidly than ER mRNA and protein expression (at 24 h and 48 h, respectively) in most uterine cells. However, the timing and extent of the estradiol response depended partly upon cell type (epithelial, stromal, or myometrial), cell region (luminal, superficial, middle, or deep endometrial or myometrial), adjacent cells, and prior progesterone treatment. For example, PR mRNA up-regulation was prolonged in middle and deep endometrial stroma, but increases in PR protein expression were highest in superficial and middle endometrial compartments, including the luminal epithelium. The luminal epithelium and myometrium were unique in that estradiol failed to up-regulate ER gene expression within them. ER mRNA levels rose within these compartments only when estradiol followed steroid hormone treatment designed to induce an artificial estrous cycle (estradiol-progesterone-estradiol [EPE] treatment). The EPE treatment also augmented the rise in ER mRNA concentrations within stromal cells compared to estradiol treatment alone. Within uterine cell compartments, subpopulations of adjacent cells showed distinct estradiol responses, e.g., very high levels of ER and PR gene expression within stromal cells directly underlying glandular epithelial cells. Because the estradiol response did not always correlate with initial ER protein levels and was partly dependent upon cell compartment and adjacent cells, we must conclude that direct transcriptional and/or posttranscriptional actions of estradiol cooperate with other cellular and paracrine regulatory factors to regulate ER and PR gene expression and, thus, the steroid responsiveness of uterine cells.

Hormone receptors in vestibular schwannomas.

The possibility that steroid hormones play a role in vestibular schwannoma proliferation has been suggested by a number of investigators. There is conflicting information about the presence of steroid hormone receptors in these tumors. The aim of this study was to examine the expression of androgen, progesterone, glucocorticoid and estrogen receptor messenger ribonucleic acid levels (mRNA) in twenty-one vestibular schwannomas by either Northern blot analysis or the polymerase chain reaction (PCR). Glucocorticoid receptor mRNA was expressed in all twenty-one tumors examined. Only two male specimens were positive for androgen receptor mRNA expression by PCR-Southern blot analysis. Thirty-three percent of the schwannomas (7/21) showed a strong band for progesterone receptor mRNA by PCR-Southern blot analysis; there were an equal number of males and females in this group. Estrogen receptor mRNA levels were undetectable in all tumors examined by PCR-Southern blot analysis. These studies suggest that the pattern of steroid receptor expression is different in schwannomas than in meningiomas. Individual vestibular schwannomas need to be examined for their steroid receptor mRNA expression mRNA expression to know whether they will be responsive.

Effects of melatonin on estrogen receptor expression in the forebrain of outbred (Lak.LVG) golden hamsters

Studies have shown that the pineal gland via its hormone, melatonin, induces the involution of male and female reproductive systems in seasonally reproducing animals. Melatonin has direct inhibitory effects on both hypothalamic and pituitary functions, which are also exquisitely sensitive to the feedback effects of estradiol. Since melatonin can modulate estrogen receptor (ER) expression in other tissues, immunocytochemical and ribonuclease protection analyses were used to examine the effects of 12 weeks of daily late afternoon injections of melatonin on ER protein and mRNA levels in the hypothalamus of Lak.LVG golden hamsters. Significant decreases in ER-immunoreactivity were noted in the medial preoptic area (MPOA) and bed nucleus of the stria terminalis (BNST) in response to melatonin, while other hypothalamic areas which express ER, e.g. the anterior hypothalamus, showed less dramatic changes. Hypothalamic ER mRNA was decreased in response to melatonin in both intact and ovariectomized animals by 25%. In intact, cycling female hamsters, there was a significant reduction in uterine weight after melatonin treatment. These results suggest that melatonin exerts its anti-reproductive effects in hamsters by modulating ER levels in neurons of the MPOA and BNST, thereby influencing steroid feedback mechanisms.

Transcriptional and post-transcriptional regulation of rainbow trout estrogen receptor and vitellogenin gene expression

Estrogen receptor (ER) and vitellogenin (Vg) gene expression are strongly up-regulated by estrogens in rainbow trout liver. In this paper, we have used primary cultured hepatocytes to examine the mechanisms implicated in estrogen regulation of ER and Vg gene expression. Treatment of hepatocytes with 1 μM estradiol (E2) led to a rapid increase in ER mRNA level (15 fold) followed by Vg mRNA induction. Transcription rate and mRNA half-life determination carried out in the presence or absence of E2, demonstrated that E2 increases both the ER and Vg gene transcriptional activity and mRNA stability (ca. 3 fold). The effect of E2 was inhibited by an excess of antiestrogens, showing that E2-stimulation of ER and Vg mRNA level is mediated by the estrogen receptor. Our data show that ER and Vg genes have different hormonal sensitivity. In fact, the Vg gene required a higher concentration of E2 to be stimulated compared to the ER gene. Examination of the mechanisms involved in post-transcriptional regulation of ER mRNA showed that the setting up and maintenance of this regulation process implies the estrogen receptor and the general translational activity within the cells, suggesting that E2-stabilization of ER mRNA depends on the synthesis of an estrogen-dependent protein. However, the cis and trans elements involved in this E2-stabilization process remain to be identified.

Up-regulation of estrogen receptor mRNA and estrogen receptor activity by estradiol in liver of rainbow trout and other teleostean fish

Injection of estradiol (E 2) into immature rainbow trout resulted in the induction of the hepatic vitellogenin gene mediated by the nuclear estrogen receptor (ER). Liver ER mRNA rose markedly on E 2 treatment in three groups of trout kept at different temperatures. Only in the group kept at 4 °C did the total cellular ER, as measured by [ 3H]estradiol-binding activity in nuclear and cytosol fractions, parallel the ER mRNA level. In fish kept at 9 °C and 15 °C, the ratio of total ER activity to ER mRNA fell during chronic E 2 treatment, probably reflecting translational or post-translational control mechanisms. Upregulation of ER mRNA also occurred in sea raven, sculpin, winter flounder, and Atlantic salmon after E 2 treatment. Intrahepatic ER activity rose proportionately in Atlantic salmon kept at 6–9 °C but not in sea raven, sculpin, or flounder. We conclude that the regulation of ER expression in teleosts is complex and includes transcriptional, translational, and post-translational elements and is influenced by environmental temperature.

Role of the pituitary in tumor promotion with ethinyl estradiol in rat liver.

Synthetic estrogens act as tumor promoters in rat liver. Because estrogen treatment markedly increases the secretion of pituitary prolactin, also shown to be a tumor promoter in rat liver, the possibility of a pituitary influence in estrogen promotion was investigated in Wistar rats. In diethylnitrosamine (DEN)-initiated hypophysectomized (hx) female rats, 24 weeks of ethinyl estradiol (EE) administration (500 microg/kg/d, intraperitoneally) did not increase the number of hepatocyte nodules and did not induce hepatocellular carcinoma (HCC) in a 2-year study. Very few placental forms of glutathione-S-transferase (GST-P)-positive foci were observed at the end of EE administration. Estrogen receptor (ER) messenger RNA (mRNA) levels in hx females were 20% of the levels in intact females. EE administration (range, 160-210 microg/kg/d, subcutaneous release pellets) to DEN-initiated intact males and females increased the number and size of hepatocyte foci. A significant increase in HCC frequency was observed in EE-treated females compared with females receiving sham-release pellets, and the latency period for HCC induction was decreased by EE in both males and females. Inhibition of prolactin (PRL) secretion by bromocriptine (Brc) (ParlodelLAR, slow intramuscular release vehicles) during EE treatment decreased the number of foci without affecting their size and markedly prolonged the latency period in both sexes. EE treatment also significantly increased the expression of c-myc, and c-jun, enhanced the levels of growth hormone receptor (GHr) mRNA in females and the levels of ER mRNA in males and "feminized" the expression of the GH-regulated genes cytochrome P450 (CYP), 2C11, CYP 2C12, and GHr in male liver. Brc administration decreased the mRNA levels of the female-predominant CYP 2C12 in EE-treated males but otherwise had no effects. In conclusion, a decreased promotive effect of EE was obtained by decreasing the PRL levels, indicating that estrogens exert at least part of their promotion effects indirectly, by increasing the levels of pituitary PRL.

Role of the pituitary in tumor promotion with ethinyl estradiol in rat liver

Synthetic estrogens act as tumor promoters in rat liver. Because estrogen treatment markedly increases the secretion of pituitary prolactin, also shown to be a tumor promoter in rat liver, the possibility of a pituitary influence in estrogen promotion was investigated in Wistar rats. In diethylnitrosamine (DEN)-initiated hypophysectomized (hx) female rats, 24 weeks of ethinyl estradiol (EE) administration (500 microg/kg/d, intraperitoneally) did not increase the number of hepatocyte nodules and did not induce hepatocellular carcinoma (HCC) in a 2-year study. Very few placental forms of glutathione-S-transferase (GST-P)-positive foci were observed at the end of EE administration. Estrogen receptor (ER) messenger RNA (mRNA) levels in hx females were 20% of the levels in intact females. EE administration (range, 160-210 microg/kg/d, subcutaneous release pellets) to DEN-initiated intact males and females increased the number and size of hepatocyte foci. A significant increase in HCC frequency was observed in EE-treated females compared with females receiving sham-release pellets, and the latency period for HCC induction was decreased by EE in both males and females. Inhibition of prolactin (PRL) secretion by bromocriptine (Brc) (ParlodelLAR, slow intramuscular release vehicles) during EE treatment decreased the number of foci without affecting their size and markedly prolonged the latency period in both sexes. EE treatment also significantly increased the expression of c-myc, and c-jun, enhanced the levels of growth hormone receptor (GHr) mRNA in females and the levels of ER mRNA in males and "feminized" the expression of the GH-regulated genes cytochrome P450 (CYP), 2C11, CYP 2C12, and GHr in male liver. Brc administration decreased the mRNA levels of the female-predominant CYP 2C12 in EE-treated males but otherwise had no effects. In conclusion, a decreased promotive effect of EE was obtained by decreasing the PRL levels, indicating that estrogens exert at least part of their promotion effects indirectly, by increasing the levels of pituitary PRL. (Hepatology 1996 Oct;24(4):849-54)

Estrogenic effect on the expression of estrogen receptor, COUP-TF, and lactoferrin mRNA in developing mouse tissues.

We have previously demonstrated that lactoferrin (LF) is a major estrogen-inducible protein in the mouse uterus. The increase of LF mRNA after estrogen treatment (> 300 fold) is the result of a complex interplay among transcription factors acting on the estrogen response element (ERE) of the LF gene. Two transcription factors-the estrogen receptor (ER) and the chicken ovalbumin upstream promoter transcription factor (COUP-TF)-play opposing roles in the estrogen responsiveness of the LF gene promoter-reporter constructs in transiently transfected human endometrial carcinoma cells. The ratio of ER/COUP-TF in the transfected cells appears to be critical for estrogen-stimulated LF gene promoter activity (Liu et al, 1993). In the current study, ER and COUP-TF mRNA levels are examined and related to LF mRNA expression in various mouse tissues, including the developing uterus with/without estrogen stimulation. Results show that LF mRNA and protein are expressed in various tissues during development, but the potent synthetic estrogen, diethylstilbestrol (DES), does not increase LF mRNA expression in nonreproductive tissues such as liver, spleen, and lung. In contrast, in developing neonatal reproductive tract tissues, DES increases LF mRNA and protein expression as previously reported in immature and mature uterine tissues. DES, however, did not affect ER and COUP-TF expression in developing uterine tissues. Although the uterus has a high ratio of ER/COUP-TF as compared to other tissues examined, COUP-TF may not be the only regulator for LF gene expression in this particular tissue since COUP-TF remains constant during development and following DES treatment. These data point to the complexity of differential expression of LF gene in estrogen responsive and nonresponsive tissues during development.

Nerve growth factor (NGF) regulation of estrogen receptors in explant cultures of the developing forebrain.

Estrogen profoundly affects the organization of the nervous system. Its receptor, a nuclear transcription factor, is widely expressed in the developing forebrain. Earlier work established that forebrain estrogen target neurons coexpress nerve growth factor (NGF) receptors and receptor mRNA. The present study examined the regulation of forebrain estrogen receptors by NGF, using organotypic cultures of the developing cerebral cortex and basal forebrain. NGF significantly increased nuclear estrogen binding in cortical but not basal forebrain explants. Both cortical and basal forebrain explant cultures express the NGF receptor, TrkA. However, our earlier observation that developing cortical neurons, unlike basal forebrain neurons, widely coexpress mRNAs for NGF and its cognate receptors, suggests that in the present study cortical neuronal responses to exogenous NGF may have been primed by autocrine mechanisms. Alterations in nuclear estrogen binding but not estrogen receptor mRNA levels suggests that NGF may regulate cortical estrogen receptors posttranscriptionally.

Inhibition of Prolactin Receptor Gene Expression by 2,3,7,8-Tetrachlorodibenzo-p-dioxin in MCF-7 Human Breast Cancer Cells

Treatment of MCF-7 human breast cancer cells with 10 nM2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) did not decrease prolactin receptor (PRLR) binding. In contrast, PRLR mRNA levels were significantly decreased within 12 h after treatment with TCDD and persisted for up to 48 h. The effects of TCDD on PRLR mRNA levels were inhibited by the aryl hydrocarbon (Ah) receptor antagonist α-naphthoflavone and were not observed in Ah nonresponsive benzo[a]pyrene-resistant MCF-7 cells. These results suggest that the effects of TCDD were mediated through the Ah receptor. After treatment of MCF-7 cells with 10 nM17β-estradiol (E2), there was a 2.3-fold increase in PRLR mRNA levels, and in cells cotreated with E2 plus TCDD, there was a 72% decrease in E2-induced PRLR mRNA levels. Previous studies have showed that TCDD also effects estrogen receptor (ER) binding and mRNA levels through the aryl hydrocarbon receptor pathway; however, the effects of TCDD on PRLR levels and binding in MCF-7 cells were different from those previously observed for ER.

Developmental profile and regulation of estrogen receptor (ER) mRNA expression in the preoptic area of prenatal rats

Estrogen, derived from circulating testosterone, masculinizes the developing preoptic area. Expression of estrogen receptors (ERs) within the preoptic area is one requirement for a possible direct action of estrogen in the process of sexual differentiation of this brain region. Using a 35S-labeled riboprobe and in situ hybridization to detect ER mRNA on both film and emulsion-coated slides, we were able to detect ER mRNA within the rat preoptic area by embryonic day 18 (ED 18), coincident with the reported onset of the critical period for testosterone-dependent masculinization of this region. ER mRNA increased significantly between ED 18 and 19 in both sexes, and continued to increase through postnatal day 0 (PND 0 = day of birth) in females, but not males. ER mRNA levels were not significantly greater in females than in males until PND 0. The lack of a sex difference in ER mRNA prenatally, however, appears to be due to an effect of intrauterine neighbors. ER mRNA levels in ED 20 embryos were relatively high in females with female-only neighbors, whereas ER mRNA levels were relatively low, and comparable to males, when the in utero neighbors included one or more males. Treatment of pregnant dams with diethylstilbestrol or with tamoxifen did not significantly alter ER mRNA levels in the preoptic area of the embryos. Although these results suggest that ER mRNA expression is subject to hormonal regulation prenatally, the relevant hormone was not identified.

Developmental profile and regulation of estrogen receptor (ER) mRNA expression in the preoptic area of prenatal rats

Estrogen, derived from circulating testosterone, masculinizes the developing preoptic area. Expression of estrogen receptors (ERs) within the preoptic area is one requirement for a possible direct action of estrogen in the process of sexual differentiation of this brain region. Using a 35S-labeled riboprobe and in situ hybridization to detect ER mRNA on both film and emulsion-coated slides, we were able to detect ER mRNA within the rat preoptic area by embryonic day 18 (ED 18), coincident with the reported onset of the critical period for testosterone-dependent masculinization of this region. ER mRNA increased significantly between ED 18 and 19 in both sexes, and continued to increase through postnatal day 0 ( PND 0 = day of birth) in females, but not males. ER mRNA levels were not significantly greater in females than in males until PND 0. The lack of a sex difference in ER mRNA prenatally, however, appears to be due to an effect of intrauterine neighbors. ER mRNA levels in ED 20 embryos were relatively high in females with female-only neighbors, whereas ER mRNA levels were relatively low, and comparable to males, when the in utero neighbors included one or more males. Treatment of pregnant dams with diethylstilbestrol or with tamoxifen did not significantly alter ER mRNA levels in the preoptic area of the embryos. Although these results suggest that ER mRNA expression is subject to hormonal regulation prenatally, the relevant hormone was not identified.

Influence of blockers for the estrogen receptor (ER) and type 1 IGF-receptor on the levels of ER, ER mRNA and IGF-I mRNA in the rat uterus.

The effects on the uterine concentration of the estrogen receptor (ER), ER mRNA and IGF-I mRNA were monitored in ovariectomized (OVX) rats treated with blockers for the estrogen receptor (ER) (ICI 182780) or the type 1 IGF receptor (H-1356), alone or in combination with estradiol (E2). The hormone-mediated increase in uterine wet weight after E2 treatment was prevented by a simultaneous injection of ICI 182780. The levels of IGF-I mRNA and ER mRNA increased in rats given estradiol (E2), but not in those also receiving ICI 182780. The level of uterine ER was decreased in all animals that received ICI 182780. H-1356 (H) given as a continuous infusion decreased the estradiol-induced increase in uterine wet weight 24 h after a single E2 injection, but not 48 h after the first (of two) E2 injections. The IGF-I mRNA was increased in the E2 treated group after 24 h, and in both the E2 and H + E2 treated groups after 48 h. The ER mRNA was increased in the E2 treated group after 24 h, but not after 48 h. When H-1356 was given as two single injections and estradiol was or was not administered together with the second injection, the uterine wet weight was not increased as in the group receiving E2 only. The levels of IGF-I mRNA as well as ER mRNA were increased both after E2 and H + E2 treatment. In conclusion, the uterine growth stimulated by E2 was prevented by simultaneous treatment with ICI 182780. In addition a type 1 IGF-receptor blocker, which is an IGF-I analogue inhibiting the autophosphorylation of the receptor, also prevented the hormone-induced uterine growth after 24 h, but not after 48 h, when given as a continuous infusion. The results imply that the growth process stimulated by estradiol utilizes the ER receptor, and also the type 1 IGF-receptor during the first 24 h after E2 treatment.

In utero and lactational exposure of female Holtzman rats to 2,3,7,8-tetrachlorodibenzo-p-dioxin: modulation of the estrogen signal.

The environmental pollutant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; dioxin) induces severe reproductive defects in male rats when exposure occurs in utero and during lactation. Yet there is currently a paucity of information regarding the effects of this exposure paradigm in females. In the current study, we examine the effects of TCDD during fetal and perinatal development on the estrogen-signaling system in peripubertal female rats. Pregnant Holtzman rats were given 1 microgram/kg TCDD or vehicle control by gavage on gestational Day 15. Body weights were reduced, though not significantly, on postnatal Day 21. While ovarian and uterine wet weights were not increased by TCDD exposure, the percentage of body weight attributed to the ovary was increased significantly. Through use of ribonuclease protection and gel-shift assays, exposed females were compared with nonexposed counterparts for estrogen receptor (ER) mRNA and DNA-binding activity in the following tissues: hypothalamus, pituitary (mRNA only), uterus, and ovary. ER mRNA levels increased in the hypothalamus, uterus, and ovary, and decreased in the pituitary. The results of the DNA-binding assays paralleled the mRNA results in the uterus, while DNA-binding activity was decreased in the hypothalamus and was unchanged in ovarian protein extracts. Circulating concentrations of estrogen were significantly lower in TCDD-exposed rats than in controls. These data suggest that the decrease in serum estrogen may be a cause of the alterations in ER mRNA; the changes in ER DNA-binding activity may indicate alterations in either translation or posttranslational receptor processing. Overall, this study shows that TCDD may act systemically in this model, and these effects should not necessarily be characterized as antiestrogenic.

Effect of gonadotropin-releasing hormone on estrogen receptor messenger ribonucleic acid level in perifused pituitary cells.

1. We examined the potential effect of GnRH pulses on pituitary estrogen receptor mRNA level. 2. The treatment of perifused pituitary cell aggregates with four hourly pulses of GnRH (10nM/1 min/h) resulted in a marked increase in the steady-state level of ER mRNA (25% vs unstimulated control, n = 3). 3. No changes were observed for the LH beta mRNA. Data suggest, for the first time, that a cross-talk between the GnRH and nuclear ER may occur in the gonadotrope cells.