Distribution Of Estrogen Receptor Alpha Research Articles

Immuno-localization of estrogen receptor (ER) and progesterone receptor (PR) in the buffalo ovary in relation to their plasma hormonal levels

In this study, we examined the tissue distribution of estrogen receptor alpha (ERα) and progesterone receptor (PR) in different compartments of the buffalo ovary during follicular and luteal phases of the estrous cycle. The receptors were localized by immunohistochemistry. Image analysis was done to quantify the immune reactivity. ERα was localized in various cell types of buffalo ovaries differentially during follicular and luteal phases of the estrous cycle. Immunoreactivity of ERα was detected in the primordial, primary, secondary and tertiary follicles, atretic follicles, in cells of the deep and superficial stroma, and the tunica albuginea. Specific immunostaining was observed with anti-ERα antibodies in the nuclei of follicular cells/granulosa cells and theca cells. No reaction was observed in the ovarian surface epithelium. In the growing follicle and secondary follicle, the immunoreaction for these receptors was strong. While in the tertiary follicles weak immunoreactions were recorded in the granulosa cells and theca cells. The progesterone receptors (PR) as revealed by immunohistochemistry were localized in the nuclei of different groups of ovarian cells. It was detected in the primordial, primary, secondary and tertiary follicles, atretic follicles, in cells of the deep and superficial stroma, and the tunica albuginea and surface epithelium. PR was localized in follicular cells of preantral and antral follicles, the stroma of the ovary, endothelial cells of blood vessels. PR positivity was found in one or two granulosa cells of primordial and primary follicles, with moderate immunoreaction, but no staining in oocytes. In the antral follicles, both granulosa cells, as well as theca cells, were immunostained for PR. In the obliterative atretic follicles, the invading stromal cells were highly positive for PR. Follicular cells of the primordial follicle and granulosa cells and theca cells of tertiary follicles had statistically higher percentage positive cells in the follicular phase as compared to the luteal phase. No staining was observed in the negative controls.

Expression of estrogen receptor alpha in developing brain, ovary and shell gland of Gallus gallus domesticus: Impact of stress and estrogen

Estrogen plays a central role in the control of reproductive behaviour and in the regulation of neuroendocrine system. To elucidate the mechanism by which it controls the stress-modulated functions, it is important to understand how estrogenic effects are mediated. The distribution of estrogen receptor alpha (ERα) protein in brain, ovary and shell gland of chickens has been investigated. Immature chickens were taken and randomly divided into four groups. First group served as control (C), second water deprived for two days (WD), third treated with estradiol benzoate (E) and fourth treated with estradiol benzoate followed by water deprivation during last two days of treatment (E + WD). The dose of estradiol benzoate administered for 15 days was 0.5 mg/100 g. Immuno-fluorescent localization demonstrated the presence of ER alpha (ERα) in hypothalamic area, principally the pre-optic area on estrogen administration. However, ERα expression in brain decreased on water deprivation. ERα expression was observed highly in granulosa and thecal cells of E and E + WD ovary compared to immature while least on water deprivation. Similarly, ERα expression was abundant in tunica muscularis and epithelial cells of E and E + WD shell gland while lesser in WD. Plasma estradiol significantly increased in E and E + WD group while decreased on water deprivation. Plasma corticosterone decreased significantly in E and E + WD group while increased in WD. Therefore, we conclude that administration of estradiol in stress decreases corticosterone induced sensitivity mediated by an increased expression of ERα in brain, ovary and shell gland. It appears that negative feedback regulation is involved between HPA-axis and HPG- axis during stress in chickens.

Immunohistochemical localization of estrogen receptor alpha (ERα) in the oviduct of Indian buffalo during follicular and luteal phases of estrous cycle.

The localization and distribution of estrogen receptor alpha (ERα) in different segments of oviduct of buffalo during follicular and luteal phases of estrous cycle were investigated using immunohistochemistry. Tissue samples from the different segments of oviduct from 12 buffaloes (six each during follicular and luteal phases of estrous cycle) were collected from slaughter house after assessing the gross morphology of ovaries. In addition, blood samples were collected from the animals before slaughter to estimate levels of estrogen and progesterone hormones. The tissue distribution of estrogen receptor was determined by immunohistochemical technique using one-step polymer HRPO staining system. The estrogen receptor was localized in the lamina epithelialis, propria submucosa, tunica muscularis, and tunica serosa. The maximum localization was observed in the lamina epithelialis, where both ciliated and secretory cell types were positive for ERα. Percentage of positive cells varied during the follicular and luteal phases of estrous cycle. The lining epithelium of oviductal glands was also intensely positive for ERα. No immunostaining was observed in any tunic of the oviduct when the primary antibody was replaced by antibody diluent or buffer, and it served as negative control. The data showed that highest immune positive cells were observed in the ampulla region of the oviduct and these cells were lowest in the utero-tubal junction (p < 0.05). Infundibulum, ampulla, and isthmus showed a higher percentage of ERα-positive cells during follicular phase of estrous cycle as compared with those of the luteal phase of estrous cycle (p < 0.05). There was no significant difference in the percentage positive cells during the two phases of estrous cycle in the utero-tubal junction. Immunogold labeling with anti-ERα antibody confirmed the findings of immunohistochemical study at subcellular level. The higher expression during the follicular phase was directly correlated with the level of estrogen hormone.

Immuno Localization of Estrogen Receptor (ERα) and Progesterone Receptor (PR) in Uterus of Buffalo during Follicular and Luteal Phases of Estrous Cycle

The tissue distribution of estrogen receptor alpha (ERα) and progesterone receptor (PR) was examined using immunohistochemical technique. Image analysis was done to quantify the immune reactivity. The ERα and PR was localized in luminal epithelium, glandular epithelium, stromal cells, endothelial cells and myometrium and few cells in perimetrium. The immuno staining was observed in the nuclei of cells, however, faint cytoplasmic staining for PR was also observed. Variations were seen in the different tissue compartments of the uterus and during the different phases of the estrous cycle. Significantly higher number of ERα positive cells was observed in lamina epithelialis as compared to stromal cells and smooth muscle cells in myometrium. Significantly higher percentage of ERα positive cells was observed in the lamina epithelialis and lining epithelium of endometrial glands during follicular phase as compared to the luteal phases of estrous cycle (P < 0.05). Higher number of PR positive cells was observed in lamina epithelialis as compared to stromal cells and smooth muscle cells in myometrium (P < 0.05). Higher percentage of PR positive cells was observed in the lamina epithelialis during follicular phase as compared to the luteal phases of estrous cycle (P < 0.05). There was no significant difference in the immuno reactivity in stromal cells, lining epithelium of endometrial glands and smooth muscle cells of myometrium during the phases of estrous cycle. The study concluded that ERα and PR expressions were higher during follicular phase as compared to the luteal phase.

Gender Difference in Distribution of Estrogen and Androgen Receptors in Intestinal-type Gastric Cancer

Gender difference in survival of patients with gastric cancer is not well investigated. The aim of this study was to analyze the gender-related distribution of estrogen receptor alpha (ERα) and androgen receptor (AR) in the epithelium and stroma of intestinal-type gastric cancer. Immunohistochemical analysis was performed in 60 patients (42% females). In gastric cancer patients, frequency of ERα-positive cells was lower in epithelium than in healthy individuals, but not significantly. In stroma and epithelium, AR-positive cells were absent from samples of women with T1 and T2 stage disease, while in men, their frequency was significantly increased in stroma of those with T3 and T4 stages and was significantly higher compared to women. AR-positive cells in stroma were fibroblasts, myofibroblasts and mast cells. To our knowledge, this study is the first to show gender differences in the distribution and frequency of AR-positive cells in neoplastic stroma of gastric cancer.

Effect of anti-PMSG on distribution of estrogen receptor alpha and progesterone receptor in mouse ovary, oviduct and uterus.

It is well established that estrogen and progesterone are critical endogenous hormones that are essential for implantation and pregnancy in females. However, the distribution of estrogen receptor α (ERα) and progesterone receptor (PR) in female reproductive tracts is elusive. Herein, we report that after serial treatments with pregnant mare's serum gonadotrophin (PMSG) with or without anti-PMSG (AP), mice could regulate the distribution of ERα and PR in the murine ovary, oviduct and uterus and the level of estradiol in serum. ERα and PR regulation by PMSG and anti-PMSG was estrous cycle-dependent and critical for promoting the embryo-implantation period. Furthermore, our results suggested that AP-42 h treatment is more effective than the other treatments. In contrast, other treatment groups also affected the distribution of ERα and PR in mouse reproductive tracts. Thus, we found that anti-PMSG has the potential to restore the distribution of ERα and PR, which could effectively reduce the negative impact of residual estrogen caused by the normal superovulation effect of PMSG in mice.

Distribution of estrogen receptor alpha and progesterone receptor, and leukocyte infiltration in the cervix of cyclic bitches and those with pyometra

The objectives were to localize estrogen receptor alpha (ERα) and progesterone receptor (PR), and enumerate leukocyte infiltration in cervical tissue of normal bitches during various stages of the estrous cycle (n = 35), as well as in those developing open (n = 22) or closed-cervix pyometra (n = 19). Each pyometra group was subdivided into anestrus and diestrus. Cervical tissues were collected after ovariohysterectomy. Receptor expressions were determined by immunohistochemistry and leukocyte infiltration was evaluated in histological sections stained with haematoxylin-eosin. The assessment was performed in two parts of cervical sections: the uterine part in four tissue layers (surface epithelium (SE), lamina propria (LP), glandular epithelium (GE), and tunica muscularis (M)), and the vaginal part in three layers (SE, LP and M). An immunohistochemical total score consisted of the addition of both the intensity and proportional scores. The ERα and PR scores differed between groups (P < 0.05) and between layers (P < 0.05), but were not significantly different between uterine and vaginal parts. The ERα score was lowest in the open-cervix pyometra bitches at anestrus and in closed-cervix pyometra bitches at diestrus. For all types of immune cells, there were no significant differences among stages of the estrous cycle in normal bitches, whereas neutrophils were lower in both sub-groups of closed-cervix versus open-cervix pyometra (P < 0.05). In conclusion, distributions of ERα and PR were similar along the longitudinal axis of the canine cervix. We inferred that cervical dilation in normal bitches and bitches with uterine pathology was likely controlled by different mechanisms. Receptor expressions were influenced by stage of the estrous cycle in normal bitches, whereas neutrophil infiltration in cervical tissue appeared to be involved in cervical dilation in bitches with pyometra, regardless of estrous stages.

Paradoxial changes in the expression of estrogen receptor alpha in breast cancer multicellular spheroids

Multicellular spheroids are excellent models for the analysis of cancer behavior. Just like small avascular tumors, they present a marked zonal heterogeneity which influences gene expression and thus, growth and response to chemotherapy. In the present paper, we sought to analyze the effects of three-dimensional culture in the expression and distribution of estrogen receptor alpha. Using MCF-7 breast cancer cells, we found that multicellular spheroids in estrogen-containing medium presented a paradoxical regulation of estrogen receptor alpha, with a decrease in protein expression and a marked increase in mRNA steady-state levels. Immunohistochemistry showed that only sparse cells in the periphery of the spheroid expressed estrogen receptor, in sharp contrast with progresterone receptor, which was more extensively expressed and HIF-alpha, which was expressed in the central core of the spheroid. This could mean that both hypoxia and ERA activation by estrogen participate in the expression heterogeneity of this hormone receptor in breast cancer These results are important to considerate in the analysis and interpretation of immunohistochemistry of ERA and downstream targets in samples of solid tumors.

Social environment alters central distribution of estrogen receptor alpha in juvenile prairie voles

It is well established that social environment, particularly isolation, has a significant impact on social behaviors and neuroendocrine responses. Estrogen receptor alpha (ERα) expression in limbic structures and associated nuclei is related to the display of social behaviors. We hypothesized that the stress of isolation would cause changes in the pattern of ERα expression in the brain. Using a highly social (typically monogamous and biparental) rodent species, the prairie vole ( Microtus ochrogaster), we housed juvenile voles with a sibling, stranger or in isolation for either 4 days or 21 days. Housing manipulations began following weaning from parents and group housed siblings. Rodents may be especially sensitive to manipulations of their social environment during this juvenile period. In particular, female prairie voles are induced ovulators, reliant upon exposure to an unrelated male (male urine) to become reproductively active. ERα immunoreactivity was quantified in the medial preoptic area (MPOA), bed nucleus of the stria terminalis (BST), ventromedial nucleus of the hypothalamus (VMH) and medial amygdala (MeA). Significantly fewer ERα immunoreactive (ERα-ir) cells were labeled in the MPOA and BST of females isolated for 21 days compared with stranger housed females. Non-significant differences were shown in the VMH and MeA of females. No differences were found in voles isolated for 4 days. These results suggest that female prairie voles may be more sensitive than males to manipulations of their social environment during the juvenile period.

Distribution of estrogen receptor alpha mRNA in the brain and inner ear of a vocal fish with comparisons to sites of aromatase expression

Among vertebrates, teleost fish have the greatest capacity for estrogen production in the brain. Previously, we characterized the distribution of the estrogen-synthesizing enzyme aromatase in the brain of the midshipman fish. Here, we investigated the distribution of estrogen receptor alpha (ERalpha). A partial cDNA of ERalpha was cloned and used to generate midshipman-specific primers for RT and real-time PCR which identified transcripts in liver and ovary, the CNS, and the sensory epithelium of the main auditory endorgan (sacculus). In situ hybridization revealed abundant expression throughout the preoptic area, a vocal-acoustic site in the hypothalamus, amygdala homologs of the dorsal pallium, the pineal organ, the inner ear, the pituitary, and the ovary. Weaker expression was found in the midbrain's nucleus of the medial longitudinal fasciculus and in the dimorphic vocal motor nucleus. ERalpha expression in the pineal, gonad, and pituitary axis may function to time seasonal abiotic cues to reproductive state, while expression in the vocal motor and auditory systems support neurophysiological evidence for estrogen as a modulator of vocal motor and auditory encoding mechanisms in midshipman fish. While ERalpha is restricted to specific nuclei, aromatase expression is abundant in glial cells throughout the entire forebrain, and high in midbrain and hindbrain - spinal vocal regions. The only site of aromatase-containing neurons is in the peripheral auditory system, where it is localized to ganglion cells in the auditory nerve. Estrogen production proximal to ERalpha-positive neurons may provide for focal sites of estrogen effects on reproductive-, vocal-, and auditory-related neurons.

Intraspecific variation in estrogen receptor alpha and the expression of male sociosexual behavior in two populations of prairie voles

Estrogen (E) regulates a variety of male sociosexual behaviors. We hypothesize that there is a relationship between the distribution of estrogen receptor alpha (ERα) and the degree of male social behavior. To test this hypothesis, ERα immunoreactivity (IR) was compared in prairie voles ( Microtus ochrogaster) from Illinois (IL), which are highly social, and Kansas (KN), which are less social. The expression of androgen receptors (AR) in males also was compared between populations. The expression of ERα and AR were compared in brains from KN and IL males and females using immunocytochemistry (ICC). There were significant intrapopulational differences, with males expressing less ERα-IR than females in the medial preoptic area, ventromedial nucleus, ventrolateral portion of the hypothalamus, and bed nucleus of the stria terminalis (BST). IL males also displayed less ERα-IR in the medial amygdala (MeA) than IL females. While IL males expressed significantly less ERα-IR in the BST and MeA than KN males, there was no difference in AR-IR. Differences in the pattern of ERα-IR between KN and IL males were behaviorally relevant, as low levels of testosterone (T) were more effective in restoring sexual activity in castrated KN males than IL males. The lack of difference in AR combined with lower expression of ERα-IR in IL males suggests that behavioral differences in response to T are associated with aromatization of T to E and that reduced sensitivity to E may facilitate prosocial behavior in males.

Distribution of Estrogen Receptor Alpha in the Dominant Follicles and Corpus Luteum at the Three Stages of Estrous Cycle in Japanese Black Cows

Distribution of estrogen receptor alpha (ERalpha) in the dominant follicle (DF) and corpus luteum (CL) at the three stages of estrous cycle in Japanese Black cows was evaluated by means of immunohistochemistry. Ovarian dynamics were observed twice daily using ultrasonography until the ovariectomy performed on Day 7 (First group, n=3), Day 10 (Second group, n=3) and Day 18 (Third group, n=3) (Day 0=estrus). Expression of ERalpha represented by immunohistological staining intensity in cells was determined using a light microscope equipped with a digital camera. A tendency toward higher expression were observed in theca interna (TI) of DF when compared with those in mural granulosa cells (mGC), antral granulosa cells (aGC) and theca externa (TE). ERalpha expression in the Third group was lower than that in the First Group in mGC, and it was also lower than that in the second group in TE (P<0.05). ERalpha expression in luteal cells was higher than those in the stromal cells in CL. No significant difference of ERalpha expression was observed within luteal or stromal cells, except in the Second group in the luteal cells, in which significantly higher expressions than that in the Third group (P<0.05) were observed. The results showed that, 1) ERalpha was present in developing DF on Day 7, early regressing DF on Day 10 and preovulatory DF on Day 18, especially in the TI, and a few were localized in the mGC, and 2) ERalpha was highly expressed in the luteal cells and the expression decreased in combination with regression of CL.

Expression of estrogen receptor alpha and 17beta-hydroxysteroid dehydrogenase 4 in the ciliary body.

To investigate the expression and distribution of estrogen receptor alpha (ERalpha) and 17beta-hydroxysteroid dehydrogenase 4 (HSD4) in bovine and monkey ciliary body. Immunohistochemical study was employed to investigate the expression of ERalpha and 17beta-HSD4. To detect ERalpha and 17beta-HSD4 transcripts, reverse transcription-polymerase chain reaction (RT-PCR) was used. In bovine and monkey ciliary body, the staining of the anti-ERalpha antibody was distributed in nonpigmented epithelium, and was not detectable in pigmented epithelium. Vessels in the stroma were also stained. The expression of 17beta-HSD4 was observed in nonpigmented and pigmented epithelium. RT-PCR revealed that the transcripts of ERalpha and 17beta-HSD4 were expressed in monkey ciliary body. This study provides evidence for the presence of ERalpha and 17beta- HSD4 in the ciliary body. The presence of ERalpha and 17beta-HSD4 suggests that estrogen may have important functions in the ciliary body.

Distribution of estrogen receptor alpha and beta immunoreactive profiles in the postnatal rat brain

The present study was conducted to identify the localization and possible contribution of the two estrogen receptor (ER) subtypes in the rat brain at postnatal (P) days 3, 7 and 14. Evaluation of the distribution of ERα and ERβ immunoreactive (ir) nuclei did not reveal gender differences at the developmental point times examined. With the exception of the cerebral cortex, the pattern of staining for these ERs was unchanged across the postnatal ages examined. The distribution of ERα-ir nuclei was wider than ERβ-ir during brain development. From P3, ERβ and ERα-ir nuclei were found in different regions of the cerebral cortex, basal forebrain, amygdala, thalamus, hypothalamus, mesencephalon, pons, cerebellum and medulla oblongata. In addition, ERα-ir nuclei were exclusively detected in the hippocampal subfields, epithalamus and in several circumventricular organs. ERα and ERβ dual immunofluorescence revealed positive nuclei in the medial part of the bed nucleus of the stria terminalis, periventricular preoptic nucleus and in caudal aspects of the ventrolateral part of the ventromedial hypothalamic nucleus. Although the functional significance of the dual expression of both ERs within the same nuclei remains unknown, it is possible that ERs play different roles in gene regulation within the same cell. The presence of ERs in diverse brain regions through early postnatal periods supports a potential role for estrogens in neural differentiation.

Estrogen receptor immunoreactivity in the adult primate brain: Neuronal distribution and association with p75, trkA, and choline acetyltransferase

The neuroactive steroid hormone, estrogen, has been implicated in both the prevention and treatment of Alzheimer's disease. Interactions between estrogen and neurotrophic systems may partially explain the beneficial effects of estrogen therapy. Previous studies have identified estrogen binding sites colocalized with neurotrophin-related proteins and mRNA within the rodent brain. Extending these studies to a model more relevant to human systems, we have mapped the distribution of estrogen receptor alpha (ER-α)-immunoreactive neurons in adult nonhuman primate brains. In addition, we used double-label immunohistochemistry to examine colocalization of ER-α with the low- and high-affinity neurotrophin receptors, p75 and trkA, and with the cholinergic marker choline acetyltransferase. Large numbers of ER-α-immunoreactive cells were detected in several amygdaloid and hypothalamic nuclei. ER-α-labeled cells were also found in the lateral septum, nucleus of the stria terminals, subfornical organ, and periaqueductal gray. Only rare, scattered ER-α-immunoreactive cells were noted in the cholinergic basal forebrain. In contrast to rodents, no cells exhibited ER-α and p75 or ER-α and trkA double-labeling. However, ER-labeled neurons in the amygdala, a region containing putative nerve growth factor-producing cells and exhibiting a role in memory, were densely and specifically invested with cholinergic terminals projecting from the basal forebrain. Estrogen-labeled neurons were also present in the lateral septal nucleus, a system that receives hippocampal inputs and projects to the neurotrophin-sensitive medial septum. Thus, interactions between neurotrophin-sensitive neurons and ER-bearing neurons exist in the primate brain, providing a potential paracrine basis for estrogen-state modulation of vulnerability to Alzheimer's disease. J. Comp. Neurol. 405:529–542, 1999. © 1999 Wiley-Liss, Inc.

Estrogen receptor immunoreactivity in the adult primate brain: Neuronal distribution and association with p75,trkA, and choline acetyltransferase

The neuroactive steroid hormone, estrogen, has been implicated in both the prevention and treatment of Alzheimer's disease. Interactions between estrogen and neurotrophic systems may partially explain the beneficial effects of estrogen therapy. Previous studies have identified estrogen binding sites colocalized with neurotrophin-related proteins and mRNA within the rodent brain. Extending these studies to a model more relevant to human systems, we have mapped the distribution of estrogen receptor alpha (ER-alpha)-immunoreactive neurons in adult nonhuman primate brains. In addition, we used double-label immunohistochemistry to examine colocalization of ER-alpha with the low- and high-affinity neurotrophin receptors, p75 and trkA, and with the cholinergic marker choline acetyltransferase. Large numbers of ER-alpha-immunoreactive cells were detected in several amygdaloid and hypothalamic nuclei. ER-alpha-labeled cells were also found in the lateral septum, nucleus of the stria terminals, subfornical organ, and periaqueductal gray. Only rare, scattered ER-alpha-immunoreactive cells were noted in the cholinergic basal forebrain. In contrast to rodents, no cells exhibited ER-alpha and p75 or ER-alpha and trkA double-labeling. However, ER-labeled neurons in the amygdala, a region containing putative nerve growth factor-producing cells and exhibiting a role in memory, were densely and specifically invested with cholinergic terminals projecting from the basal forebrain. Estrogen-labeled neurons were also present in the lateral septal nucleus, a system that receives hippocampal inputs and projects to the neurotrophin-sensitive medial septum. Thus, interactions between neurotrophin-sensitive neurons and ER-bearing neurons exist in the primate brain, providing a potential paracrine basis for estrogen-state modulation of vulnerability to Alzheimer's disease.