Effects Of Environmental Estrogens Research Articles

New frontiers of developmental endocrinology opened by researchers connecting irreversible effects of sex hormones on developing organs

In the early 1960's, at Professor Bern's laboratory, University of California, Berkeley) in the US, Takasugi discovered ovary-independent, persistent vaginal changes in mice exposed neonatally to estrogen, which resulted in vaginal cancer later in life. Reproductive abnormalities in rodents were reported as a result of perinatal exposure to various estrogenic chemicals. Ten years later, vaginal cancers were reported in young women exposed in utero to the synthetic estrogen diethylstilbestrol (DES) and this has been called the “DES syndrome”. The developing organism is particularly sensitive to developmental exposure to estrogens inducing long-term changes in various organs including the reproductive organs. The molecular mechanism underlying the persistent vaginal changes induced by perinatal estrogen exposure was partly demonstrated. Persistent phosphorylation and sustained expression of EGF-like growth factors, lead to estrogen receptor α (ESR1) activation, and then persistent vaginal epithelial cell proliferation. Agents which are weakly estrogenic by postnatal criteria may have major developmental effects, especially during a critical perinatal period. The present review outlines various studies conducted by four generations of investigators all under the influence of Prof. Bern. The studies include reports of persistent changes induced by neonatal androgen exposure, analyses of estrogen responsive genes, factors determining epithelial differentiation in the Müllerian duct, ESR and growth factor signaling, and polyovular follicles in mammals. This review is then expanded to the studies on the effects of environmental estrogens on wildlife and endocrine disruption in Daphnids.

Missing Information from the Estrogen Receptor Puzzle: Where Are They Localized in Bull Reproductive Tissues and Spermatozoa?

Estrogens are steroid hormones that affect a wide range of physiological functions. The effect of estrogens on male reproductive tissues and sperm cells through specific receptors is essential for sperm development, maturation, and function. Although estrogen receptors (ERs) have been studied in several mammalian species, including humans, they have not yet been described in bull spermatozoa and reproductive tissues. In this study, we analyzed the presence of all types of ERs (ESR1, ESR2, and GPER1) in bull testicular and epididymal tissues and epididymal and ejaculated spermatozoa, and we characterize them here for the first time. We observed different localizations of each type of ER in the sperm head by immunofluorescent microscopy. Additionally, using a selected polyclonal antibody, we found that each type of ER in bull sperm extracts had two isoforms with different molecular masses. The detailed detection of ERs is a prerequisite not only for understanding the effect of estrogen on all reproductive events but also for further studying the negative effect of environmental estrogens (endocrine disruptors) on processes that lead to fertilization.

Zeranol stimulates proliferation and aromatase activation in human breast preadipocytes.

Aromatase is a crucial enzyme for the biosynthesis of estrogens and is involved in the process of breast carcinogenesis. Concerns have been raised regarding the effects of environmental estrogens as potential regulators of aromatase expression in human breast cells. Zeranol is a non‑steroidal agent with potent estrogenic activity, which is widely used as a growth promoter for cattle in certain countries. The present study hypothesized that aromatase expression and activity may be elevated by low dose zeranol exposure, providing a source of estrogens that may stimulate cell proliferation. In the present study, primary cultured human breast preadipocytes were used as an invitro model. The effects of zeranol on cell proliferation were measured using the MTS assay, aromatase expression levels were determined by immunocytochemical staining and reverse transcription‑polymerase chain reaction, and aromatase enzyme activity and estrogen production were analyzed using corresponding assay kits. The results demonstrated that low dose zeranol (2‑50nM) was able to significantly promote cell proliferation, aromatase mRNA expression, aromatase activity and estrogen production in primary cultured human breast preadipocytes, thus suggesting that zeranol may act as an aromatase activator. The findings of the present study suggest that zeranol promotes breast cancer cell growth by stimulating aromatase activation and increasing estrogen biosynthesis in adipose tissue.

Alleviation of Drought Stress in <i>Arabidopsis thaliana</i> by 17<i>β</i>-Estradiol Application

Animal steroidal hormones, including estrogens, are being introduced into the agricultural soil and water supply from increased pharmaceutical and farm waste. Considering the current levels of xenoestrogen contamination of plant environments in view of the climate change induced drought conditions, this study was designed to understand the effect of estradiol (ES) application on Arabidopsis drought stress responses. Estradiol treatment (10 nM, 100 nM) of plants subjected to drought stress conditions by withholding water for 7 days resulted in increased tolerance to drought stress reflected in the significantly higher plant survival rates of 74% and 78%, respectively compared to control plants’ survival rates of 36% (no treatment) and 40% (mock treatment). Estradiol application significantly increased the content of glutathione, proline and H2O2 and significantly enhanced the transcription of the stress responsive genes GSTU3, GER5, HSP101, and HSP70b. A high concentration of ES (10 μM) did not protect plants against drought stress and proved to be toxic. These results provide new insight into the effect of ES on drought-stress responses in Arabidopsis with possible practical agricultural applications regarding the effect of environmental estrogens on crop plants.

Fetal and neonatal exposure to the endocrine disruptor, methoxychlor, reduces lean body mass and bone mineral density and increases cortical porosity.

Endogenous estrogen has beneficial effects on mature bone and negatively affects the developing skeleton, whereas the effect of environmental estrogens is not known. Methoxychlor (MXC) is a synthetic estrogen known as a persistent organochlorine and used as a pesticide. Methoxychlor and its metabolites display estrogenic, anti-estrogenic and anti-androgenic activity and may therefore influence bone. Fifty-eight male fetal and neonatal rats were exposed to either: a negative control (DMSO), 0.020, 100 mg/kg MXC, or 1 mg/kg β-estradiol-3-benzoate (EB; positive control). Rats were treated daily for 11 days, from embryonic day 19 to postnatal day (PND) 7 or for 4 days during the postnatal period (PND 0-7). All rats were analyzed at PND-84. Total body, femur, spine, and tibia areal bone mineral density (BMD) and content (BMC), lean body mass (LBM) and fat were measured by dual energy X-ray absorptiometry. Bone geometry and volumetric (v) BMD were measured using micro-computed tomography and biomechanical properties using three-point bending were assessed. Rats exposed to EB or MXC (at either the high and/or low dose), independent of exposure interval showed lower body weight, LBM, tibia and femur BMD and length, and total body BMD and BMC than DMSO control group (p ≤ 0.05). Methoxychlor and EB exposure increased cortical porosity compared to DMSO controls. Trabecular vBMD, number and separation, and cortical polar moment of inertia and cross-sectional area were lower due to EB exposure compared to control (p < 0.05). Early MXC exposure compromises cortical porosity and bone size at maturity, and could ultimately increase the risk of fracture with aging.

Developmental Effects of Environmental Estrogens on Subsequent Fertility and Ovarian Function.

Developmental Effects of Environmental Estrogens on Subsequent Fertility and Ovarian Function.

Effects of Environmental Estrogens on Animals in Israel: Implications for Effects on Humans

In the last decade interest has increased in the possible effects of endocrine disruptor compounds. Numerous papers have appeared as to their possible effects on humans, but definitive effects are hard to demonstrate. In the field of animal husbandry, however, the effects of the endocrine disruptors are well documented. This paper discussed the effects of the endocrine disruptors seen in animals and the possible implications for humans. The areas considered are reproductive disorders, premature udder/breast development, prolapsed oviduct/uterus, scrotal atrophy, and skewed sex ratio.

Environmentally Relevant Concentrations of 17α-Ethinylestradiol (EE2) Interfere With the Growth Hormone (GH)/Insulin-Like Growth Factor (IGF)-I System in Developing Bony Fish

The aim of this study was to evaluate whether effects of environmental estrogens on fish growth and reproduction may be mediated via modulating the growth hormone (GH)/insulin-like growth factor I (IGF-I) system. To this end, developing male and female monosex populations of tilapia were exposed to 17alpha-ethinylestradiol (EE2) at 5 and 25 ng EE2/l water from 10-day postfertilization (DPF) until 100 DPF. Under exposure to both EE2 concentrations, sex ratio shifted toward more females and body length, and weight were significantly reduced in males. The growth-reducing effect was associated with significant changes in hepatic IGF-I expression, both in males and females and with significant alterations of IGF-I mRNA and GH mRNA in the brain. The changes in IGF-I and GH mRNA were accompanied by altered estrogen receptor alpha (ERalpha) expression in brain and liver. These findings point to an influence of estrogenic exposure on the endocrine GH/IGF-I axis. In addition, the EE2 treatment resulted in significant changes of ERalpha and IGF-I expression in ovaries and testis, suggesting that the estrogens interact not only with the endocrine but also with the autocrine/paracrine part of the IGF-I system. Overall, our results provide evidence that EE2 at environmentally relevant concentrations is able to interfere with the GH/IGF-I system in bony fish and that the impairing effects of estrogens reported on fish growth and reproductive functions may rather result from a cross talk between the sex steroid and the IGF-I system than be toxicological.

Quantitative studies on the effects of environmental estrogens on the testis of the guppy, Poecilia reticulata

Estrogens that enter into the water system, such as by sewage effluent, may affect the reproductive capabilities of aquatic animals. The guppy ( Poecilia reticulata) was chosen as a model organism to test the level of disturbances in primary and secondary sexual characteristics. Male guppies were exposed to 17β-estradiol (10 and 50 ng L −1), and 17α-ethinylestradiol (10, 50 and 200 ng L −1) for 3.5 months from birth to adulthood. Specimens were euthanized and the testes removed for histological examination, applied in conjunction with stereological technique. Alterations in the quantity of sperm, decreasing intensity of body coloration, and increased body weight were generally significant only in the group exposed to 200 ng L −1 of 17α-ethinylestradiol. No significant changes were seen in the gonadosomal index (GSI). Quantification of the developmental stages of spermatogenesis indicated a trend of greater representation of later stages in specimens exposed to low doses of estrogens, and greater prevalence of early stages in those exposed to high doses of estrogens. Based on the results of this study, environmental estrogens do not seem to pose a reproductive threat to guppies. It is concluded that stereological histology can be considered a valuable tool in quantifying estrogenic effects upon fish gonads and that the guppy is perhaps not an ideal test organism for estrogenic exposure due to limited sensitivity.

Multihormonal Control of Vitellogenesis in Lower Vertebrates

The comparative approach on how and when vitellogenesis occurs in the diverse reproductive strategies displayed by aquatic and terrestrial lower vertebrates is presented in this chapter; moreover, attention has been paid to the multihormonal control of hepatic vitellogenin synthesis as it is related to seasonal changes and to vitellogenin use by growing oocytes. The hormonal mechanisms regulating vitellogenin synthesis are also considered, and the effects of environmental estrogens on the feminization process in wildlife and humans have been reported. It is then considered how fundamental nonmammalian models appear to be, for vitellogenesis research, addressed to clarifying the yolkless egg and the evolution of eutherian viviparity.

Experimental Evaluation of Vitellogenin as a Predictive Biomarker for Reproductive Disruption

Vitellogenin (VTG) synthesis in male oviparous vertebrates is used as an indicator of environmental estrogen exposure, but the relationship between elevated VTG levels and the effects of environmental estrogens on reproductive success are poorly understood. To examine whether altered VTG expression predicts reproductive impairment, we exposed medaka (Oryzias latipes) for 2 or 8 weeks posthatch to 0, 0.5, 1.0, 2.5, and 7.5 ppb of the environmental estrogen o,p'-DDT. Fish were sampled 2, 4, and 8 weeks after hatch to examine VTG expression and gonad development. After exposure, fish were transferred to clean water, grown to sexual maturity, and placed in mating pairs. We collected eggs for 7 days and scored them for fecundity (number of eggs), fertility (percent fertilized), and hatching success (percent hatched). DDT had no effect on VTG expression after a 2-week exposure, whereas all doses induced VTG after 8 weeks. At both exposure durations, the highest doses of DDT caused a female-skewed sex ratio in adults. Gonadal feminization appeared to be progressive: some ovotestes were observed after 2- or 4-week exposure to the two highest doses, but the proportion of ovaries increased after 8 weeks. Both 2- and 8-week exposures significantly reduced fertility and hatching success at all doses, with lower doses having a greater effect after longer exposure. Fertility and hatching success were more sensitive to estrogenic disruption than were gonad differentiation and vitellogenin expression. We suggest that VTG expression may be interpreted as a warning of reproductive consequences, but absence of expression cannot be interpreted as absence of consequences.

Experimental evaluation of vitellogenin as a predictive biomarker for reproductive disruption.

Vitellogenin (VTG) synthesis in male oviparous vertebrates is used as an indicator of environmental estrogen exposure, but the relationship between elevated VTG levels and the effects of environmental estrogens on reproductive success are poorly understood. To examine whether altered VTG expression predicts reproductive impairment, we exposed medaka (Oryzias latipes) for 2 or 8 weeks posthatch to 0, 0.5, 1.0, 2.5, and 7.5 ppb of the environmental estrogen o,p'-DDT. Fish were sampled 2, 4, and 8 weeks after hatch to examine VTG expression and gonad development. After exposure, fish were transferred to clean water, grown to sexual maturity, and placed in mating pairs. We collected eggs for 7 days and scored them for fecundity (number of eggs), fertility (percent fertilized), and hatching success (percent hatched). DDT had no effect on VTG expression after a 2-week exposure, whereas all doses induced VTG after 8 weeks. At both exposure durations, the highest doses of DDT caused a female-skewed sex ratio in adults. Gonadal feminization appeared to be progressive: some ovotestes were observed after 2- or 4-week exposure to the two highest doses, but the proportion of ovaries increased after 8 weeks. Both 2- and 8-week exposures significantly reduced fertility and hatching success at all doses, with lower doses having a greater effect after longer exposure. Fertility and hatching success were more sensitive to estrogenic disruption than were gonad differentiation and vitellogenin expression. We suggest that VTG expression may be interpreted as a warning of reproductive consequences, but absence of expression cannot be interpreted as absence of consequences.

Inhibition of environmental estrogen-induced proliferation of human breast carcinoma MCF-7 cells by flavonoids.

In the present study, we evaluated the individual and combined effects of environmental estrogens and flavonoids on the proliferation of human breast carcinoma MCF-7 cells. These compounds are as follows: (1) pharmaceutical chemicals such as diethylstilbestrol, 17alpha-ethynylestradiol (17ES), tamoxifen, mestranol, and clomiphene, (2) industrial chemicals such as bisphenol A (BisA), 4-octylphenol (OP), 4-nonylphenol (NP), and p,p'-biphenol, and (3) flavonoids such as daidzein (D), genistein (G), quercetin (Q), and luteolin (L). We found that nanomolar concentrations of 17ES, BisA, OP, and NP were sufficient to stimulate the proliferation of MCF-7 cells. Among them, 1 microM BisA exhibited cell proliferation-stimulating activity as strong as 10 nM 17beta-estradiol; and D and G exhibited cell proliferation-stimulating activity at 10 nM. On the other hand, Q and L exhibited cell proliferation-inhibiting activity. We also found that 10 nM flavonoids, such as D, G, Q, and L, were able to inhibit the proliferation-stimulating activity in MCF-7 cells by 1 microM environmental estrogens.

The effect of environmental estrogens on the expression of the CYP1A1, CYP1A2, and CYP3A4, CYP3A5 genes in the estrogen-dependent cell-line MCF-7

The effect of environmental estrogens on the expression of the <i>CYP1A1, CYP1A2</i>, and <i>CYP3A4, CYP3A5</i> genes in the estrogen-dependent cell-line MCF-7

Screening foods of the endangered Kakapo parrot ( Strigops habroptilus ) for oestrogenic activity using a recombinant yeast bioassay

In the past decade the possible effects of environmental oestrogens, both natural and synthetic, on the reproduction of vertebrates has become an issue of both public and scientific interest (Turner and Sharpe, 1997). While the general significance of such chemicals remains controversial there is clear evidence that, in some contexts, phytooestrogens can influence the fertility of vertebrates (Leopold et al., 1976; Shutt, 1976). While the impact of natural and synthetic environmental oestrogens may be slight, highly endangered species do represent a unique situation in which even modest reductions in the fertility of key individuals may have implications for the survival of the entire species. From these considerations it was deemed prudent to determine if oestrogenic activity could be detected in the natural and artificial foods of the endangered New Zealand nocturnal parrot, the Kakapo (Strigops habroptilus; Powlesland and Lloyd, 1994). Two issues were to be addressed: (i) whether physiologically significant levels of oestrogenic activity were detectable in the artificial (adult supplementary and chick-raising) foods of Kakapo; and (ii ) whether oestrogenic activity could be detected in either crude extracts of the Kakapo’s natural foods (i.e. native tree fruits) or chemicals isolated from these foods. A simple and robust recombinant yeast bioassay (developed by Glaxo-Wellcome) was adapted for this work. Briefly, the recombinant yeast strain used has been transformed with two genes: the human oestrogen receptor (hER) and a lacZ reporter gene located adjacent to an oestrogen response element (ERE-lacZ). Transcription of the lacZ gene in this strain is regulated through the ERE; thus the cells produce increased amounts of b -galactosidase enzyme (encoded by lacZ ) in response to any chemical that can activate the hER and cause it to bind the ERE (5). It should be emphasised that application of this bioassay requires little prior knowledge of the chemistry properties of possible oestrogenic chemicals other than requiring their solubility in 100–70% ethanol. A typical extraction procedure for the foods was as follows: weighed samples (c.100 mg) were extracted overnight (30°C on a rotating wheel) in 1·5 ml of either water alone or water with b -glucosidase (10 or 100 μg/ml). Isoflavones are present in plant cells largely as conjugated glucosides which show little oestrogenic activity: therefore b -glucosidase was added to some extraction suspensions to catalyse hydrolysis of sugar– isoflavone bonds. After adding 3·5 ml of absolute ethanol (i.e. making the extraction solvent to 70% ethanol) the samples were again extracted overnight, centrifuged and filtered through a 0·22 μm filter. Dilution series of 10 μl aliquots of the extracts were assayed as described in Routledge and Sumpter (1996). Foliage of oestrogenic clovers was used as a positive control for both the extraction and assay procedure. Purified phyto-chemicals assayed were dissolved in 96% ethanol before a dilution 1:1 series was assayed. No oestrogenic activity was detected in 70% ethanol extracts of the following adult supplementary foods: sunflower seeds, safflower seeds, walnuts, pumpkin seeds and almonds. However, very low levels of oestrogenic activity were detected in extracts of one of three commercial chick-raising products screened. The bioassay detected high levels of oestrogenic activity in 70% ethanol extracts of the positive control, clover leaves. The following chemicals, possibly present in fruits forming part of the natural Kakapo diet, were screened using the bioassay: coniferyl alcohol, gibberellins (GA9, methyl-GA9, GA4, methyl-GA4, GA3, methyl-GA3), totarol, kaurene, kaurenoic acid, phyllocladene, brassinolide. No oestrogenic activity was detected for any of these chemicals. An earlier paper (Brandt and Ross, 1948) had reported (without presenting data) oestrogenic activity for the alcohol derivative of podocarpic acid, a major component of the resin of rimu (Dacrydium cupressinum), a New Zealand forest tree that is a major natural food source for Kakapo. We confirmed this report, showing podocarpinol had oestrogenic activity comparable with genistein (c.10-fold less than 17-b estradiol) while podocarpic acid itself also had weak oestrogenic activity (c.10-fold less than 17b estradiol). The significance, if any, of this to the reproduction of Kakapo is not clear.

4-Hydroxytamoxifen-induced cytotoxicity and bisphenol A: competition for estrogen receptors in human breast cancer cell lines.

Increasing concerns over the effects of environmental estrogens on wildlife and humans have highlighted the need for screening systems to assess potentially estrogenic effects of test compounds. As a result, in vitro screening methods such as cell proliferation assays using the estrogen-responsive human breast cancer cell line, MCF-7, have been developed. The present study describes an alternative in vitro approach for the assessment of such xenoestrogens, based on estrogenic rescue of MCF-7 cells from antiestrogen-induced cytotoxicity. This method measures the ability of various estrogenic compounds to compete with a known estrogen-receptor-mediated antihormonal drug, 4-hydroxytamoxifen, using the 1-[4,5-dimethylthiazol-2-yl]-3,5-diphenylformazan (MTT) assay to assess mitochondrial activity. Because 4-hydroxytamoxifen treatment of cells results in a dramatic decrease in mitochondrial dehydrogenase activity which is directly related to their estrogen-receptor content, inhibition of this effect with estrogenic compounds represents an estrogen-receptor interaction, or estrogenic rescue. The estrogenic compounds tested include a weak xenoestrogen, bisphenol A (BPA), and two biological estrogens, 17alpha- and 17beta-estradiol. Competitive inhibition of 4-hydroxytamoxifen-induced cytotoxicity by BPA was compared to that of the biological estrogens. The results indicate that the biological estrogens can successfully compete with the antiestrogen in a dose-dependent manner. In addition, the assay is sensitive enough to detect estrogenic rescue by even the very weak xenoestrogen, BPA, albeit at high BPA concentrations. This simple in vitro method could be used as an alternative or second-line screen for potential xenoestrogens.

Elevated serum vitellogenin levels and gonadal abnormalities in wild male flounder ( Pleuronectes yokohamae) from Tokyo Bay, Japan

Concentrations of vitellogenin (VTG) in serum of wild male flounder ( Pleuronectes yokohamae) were investigated, as a biomarker for environmental estrogens in marine waters, from January 1997 to May 1998 in Tokyo Bay, Japan ( n=130) and from a reference site in Hokkaido, Japan, far from urban areas (about 40 km from Hakodate city) from March 1997 to June 1998 ( n=62). Statistically higher concentrations of serum VTG were detected in wild male flounder collected in Tokyo Bay (range: 25–2200 ng ml −1) compared to fish from Hokkaido (range: 31–91 ng ml −1) throughout the entire sampling period. Three out of 20 males collected off Haneda, in the inner part of Tokyo Bay, showed testes containing small numbers of oocytes. This intersex condition was not found among males collected from the reference site. This study suggests that the abnormally high levels of VTG and the presence of oocytes in the testis of male flounder collected from Tokyo Bay could be the effect of environmental estrogens.

Effects of environmental estrogens on tumor necrosis factor alpha-mediated apoptosis in MCF-7 cells.

Environmental estrogens represent a class of compounds which have been shown to mimic the effects or activity of the naturally occurring ovarian hormone 17beta-estradiol. Given the role of 17beta-estradiol in cell survival in a number of systems, we wished to determine if environmental estrogens protect MCF-7 cells from apoptosis. Here we demonstrate that the organochlorine pesticides o, p' DDT and alachlor, like 17beta-estradiol, have the ability to suppress tumor necrosis factor alpha (TNF)-induced apoptosis in estrogen receptor (ER)-positive MCF-7 breast carcinoma cells. These compounds, however, did not affect TNF-induced apoptosis of the ER-negative MDA-MB-231 cell line. The ability of these compounds to suppress apoptosis in MCF-7 cells was correlated with an ER-dependent increase in Bcl-2 expression. Taken together these results demonstrate that estrogenic organochlorine pesticides like o, p' DDT and alachlor may partially mimic the primary endogenous estrogen, 17beta-estradiol, and function to suppress apoptosis in ER-responsive cells.

Effect of environmental estrogens on IL-1beta promoter activity in a macrophage cell line.

Environmental estrogens or estrogen disrupters have recently received a great deal of attention because of their potential health impact on reproductive tissues. Few, if any, studies have been made on the impact of these compounds on the immune system. We sought to determine the activities of various environmental estrogens on the modulation of the interleukin-1beta (IL-1beta) gene in a model monocytic cell line, hER + IL-1beta-CAT+. This cell line stably transfected with the human estrogen receptor, and an IL-1beta promoter construct fused to the CAT reporter gene allows us to monitor the effect of estrogenic compounds on IL-1beta promoter activity. 17beta-estradiol (E2) markedly enhanced lipopolysaccharide- (LPS) induced IL-1beta promoter-driven CAT activity in a dose-dependent manner. The mycotoxins alpha-zearalenol and zearalenone both exhibited full agonist activity, but at lower potencies, with EC50 values of 1.8 and 54 nM, respectively, compared with E2 at 0.5 nM. In addition, genistein was a very low-potency agonist, having an EC50 of 1.5 microM. Similar to the E2 response, the slope factors for alpha-zearalenol, zearalenone, and genistein were close to 3.0, suggesting positive cooperativity in the estrogenic response. The activity of the mycotoxins appeared to be mediated through the estrogen receptor, since both the antiestrogens H1285 and ICI 182,780 effectively inhibited their agonist activity in a dose-dependent manner. Representative environmental estrogenic compounds both from plant and industrial sources were also tested. Unlike the mycoestrogens, none of the compounds, with the exception of genistein, synergized with LPS to enhance IL-1beta promoter activity. When tested for antiestrogenic activity, the industrial compound 4-octylphenol was able to antagonize the response to E2; however, the response was three orders of magnitude less potent than H 1285. Naringenin, a plant flavonoid, showed little or no ability to antagonize the response to E2. Overall, the results show that some environmental estrogens that display agonist activity in reproductive tissue also have an effect on IL-1 gene expression in hemopoietic-derived tissue.

Human estrogen receptor beta-gene structure, chromosomal localization, and expression pattern.

The estrogen receptor (ER) is a ligand-activated transcription factor that mediates the effects of the steroid hormone 17 beta-estradiol, in both males and females. Since the isolation and cloning of ER, the consensus has been that only one such receptor exists. The finding of a second subtype of ER (ER beta) has caused considerable excitement amongst endocrinologists. In this article, we present data regarding the genomic structure and chromosomal localization of the human ER beta gene, demonstrating that two independent ER genes do exist in the human. Furthermore, we present data regarding the tissue distribution of human ER beta, showing that this receptor is expressed in multiple tissues. For instance, ER beta is found in developing spermatids of the testis, a finding of potential relevance for the ongoing debate on the effects of environmental estrogens on sperm counts. In addition, we find ER beta in ovarian granulosa cells, indicating that estrogens also participate in the regulation of follicular growth in the human.