雌激素受体α、β在妊娠猪子宫中的表达

众所周知,子宫是一个内分泌、旁分泌和自分泌的分子作用靶器官,类固醇激素对其具有特殊的重要性。本试验通过RT-PCR、Western bloting和免疫组化技术对怀孕猪整个妊娠期（交配后10、18、32、50、71、90 d）的雌激素受体（ERα和ERβ）mRNA和蛋白质进行了定位。研究结果首次发现,猪的整个妊娠周期中都有雌激素受体蛋白存在。ERα存在于怀孕猪子宫的腔上皮（LE）、腺上皮（GE）细胞和子宫肌层的肌细胞中,而且在妊娠的不同阶段染色的强度并不相同;在怀孕猪妊娠周期内LE和GE细胞中均可检测到ERβ,但子宫肌层上仅只有少许细胞中存在ERβ,并且染色结果显示弱免疫反应。Western bloting分析结果表明,试验期内都能检测到ERα和ERβ蛋白。RT-PCR结果显示,在妊娠期的每一阶段都有ERα和ERβ的mRNA表达。以上结果表明,相对ERβ而言,妊娠猪子宫中ERα含量更丰富,且占主导地位;怀孕猪整个妊娠周期内都能发现ERα和ERβ,并且可能对雌激素细胞的增值和分化起直接作用。     

雌激素受体α在雌性大鼠脑中的表达

本试验利用免疫组织化学SP法,研究了雌激素受体α在大鼠脑中的表达.结果表明,ER_α免疫反应产物主要定位于神经元细胞核,部分区域出现阳性神经元和阳性突起,在海马CA1、CA2辐射层、齿状回、下托的分子层等出现阳性星形胶质细胞;ER_α在脑中表达广泛,免疫反应产物在端脑的隔外侧核、海马锥体细胞层、大脑皮质等,间脑的缰内侧核、下丘脑室旁核、弓状核、视上核等,中脑的动眼神经核,脑桥的脑桥核外侧部等为高密度;在端脑的尾壳核、杏仁中央核、海马CA4区,间脑的下丘脑室周核、缰外侧核等,中脑的黑质致密带,脑桥的脑桥被盖核等为较高密度;在问脑的视前室周核、丘脑后核、中脑的红核、脑桥的脑桥被盖网状核等为中等密度;在端脑的苍白球,间脑的无名质,无免疫反应产物出现.结果显示,雌激素在调节脑的认知和生殖内分泌过程中,ER_α可能起主要作用;雌激素可通过ER_α调节星形胶质细胞功能.     

雌激素β受体在大鼠睾丸中的表达与发育研究

为了研究雌激素β受体(ER-β)在不同年龄大鼠睾丸中的表达,试验采用免疫组织化学SP法对雌激素β受体在4,23,50周龄大鼠睾丸组织中的表达进行了研究。结果表明:4周龄大鼠睾丸雌激素β受体免疫反应产物表现出强阳性,主要分布在间质细胞和肌样细胞中;23周龄雌激素β受体免疫反应产物表现出中等阳性,主要出现在少量的间质细胞和大量的肌样细胞中;50周龄大鼠睾丸仅见极少数区域个别的阳性间质细胞与肌样细胞,免疫反应产物表现出弱阳性;随着大鼠年龄的增长,大鼠睾丸中的雌激素β受体的表达量呈下降趋势。     

雌激素受体在哺乳动物胎儿性腺中的分布

雌激素对胎儿性腺的发育起着重要作用,其作用是通过位于性腺的特异性受体,即雌激素受体(ER)来实现的。ER包括α(ERα)和β(ERβ)两种亚型。本文总结了此前关于两种雌激素受体亚型在胎儿性腺分布的研究,发现对于不同物种,胎儿性腺雌激素受体的分布明显不同,而且即使是在同种动物,在不同年龄段、不同生理状态、不同部位其受体的表达强度也有变化。     

山羊β雌激素受体多克隆抗体的制备及免疫组化方法的建立

本研究旨在克隆济宁青山羊雌激素受体β(ERβ)基因,进行原核表达,制备多克隆抗体,建立免疫组化分析方法,检测济宁青山羊卵巢和子宫内ERβ分布。根据GenBank发布的绵羊ERβ基因序列设计一对引物,应用RT-PCR方法从济宁青山羊卵巢组织中扩增ERβ部分基因。经双酶切和测序分析后,连接到原核表达载体pET32a(+),构建重组表达载体pET32a(+)-ERβ,转化至大肠杆菌BL21(DE3)中,IPTG诱导表达,表达产物经SDS-PAGE和Western blotting分析鉴定;经Ni-NTA纯化融合蛋白后免疫新西兰大白兔,制备多克隆抗体,并检测抗体效价及特异性;使用该抗体检测济宁青山羊卵巢和子宫组织细胞中的ERβ分布。结果表明,成功扩增出ERβ部分基因,并构建原核表达质粒,转化至大肠杆菌中表达出相对分子质量约为53ku的融合蛋白;Western blotting证明该融合蛋白能与兔抗人的ERβ多克隆抗体特异性反应。制备的多克隆抗体经ELISA检测效价达到1∶213,以此抗体代替购置的标准兔抗人ERβ抗体,进行Western blotting反应,特异性良好;使用该抗体建立免疫组化方法检测结果表明济宁青山羊卵巢颗粒细胞和子宫内膜上皮细胞、平滑肌细胞存在ERβ的分布。本研究获得特异性兔抗山羊ERβ多克隆抗体,使用该抗体建立免疫组化方法首次报道了济宁青山羊卵巢和子宫ERβ的分布表达,为进一步研究β雌激素受体的生物学功能奠定了方法学和形态学基础。     

雏鸡脑雌激素受体的表达及分布

采用免疫组化SP法研究了雌激素受体在lO日龄雏鸡脑组织内的表达，着重观察了雌激素受体在小脑、中脑、下丘脑及端脑的分布。研究表明，雌激素受体主要存在于细胞核中，少数区域仅存于胞浆或胞膜。雌激素受体在脑内分布广泛。在小脑中部皮质的颗粒层、蒲肯野氏层，中脑的中央白质、外侧丘系腹侧核、视束、后连合等区域，雌激素受体免疫反应产物为高密度；在小脑前部皮质的颗粒层，中脑中央灰质、尾侧线形核，端脑副高纹状体等区域，雌激素受体免疫反应产物为中等密度；小脑前部皮质的蒲肯野氏细胞，下丘脑视上核，端脑原始旧纹状体等区域，雌激素受体免疫反应产物为低密度。结果揭示，在鸡脑早期发育过程中，雌激素起著广泛而重要的作用。     

雌激素受体和低密度脂蛋白受体在动脉粥样硬化兔组织中的表达

用饲喂正常标准兔料的去势雌兔作为对照组,高胆固醇饮食法复制动脉粥样硬化模型。采用βactin做内参照的半定量RTPCR方法,对2组去势雌兔心脏、肝脏组织内雌激素受体α和低密度脂蛋白受体mRNA的表达进行了研究。结果表明:2种受体在兔心脏和肝脏内均有表达;对照组,雌激素受体在心脏、肝脏内的相对表达量为0.43±0.12、0.39±0.20,低密度脂蛋白受体在心脏、肝脏内的相对表达量为0.27±0.05、0.86±0.12,差异显著(P<0.01);模型组,雌激素受体和低密度脂蛋白受体在心脏、肝脏内的相对表达量分别为0.29±0.03、0.27±0.06和0.04±0.01、0.17±0.02。     

雌激素对睾丸发育及生精功能的影响

雌激素与睾丸的功能密切相关.人和哺乳动物体内雌激素的合成需芳香化酶(Ar)的参与,而雌激素与特异受体(ERα、ERβ)结合后才发挥生物学效应.在睾丸所有发育阶段,芳香化酶和雌激素受体几乎都有表达.在睾丸发育过程中,雌激素的缺乏对胎儿睾丸的发育有利.但对成年动物来说,大剂量的雌激素可以引起睾丸功能紊乱和生精障碍,而体外条件下小剂量的雌激素又能促进精原细胞的分裂、增生和分化,诱导精子发生和精子成熟,减少细胞凋亡.文章主要从睾丸内雌激素的来源、芳香化酶和雌激素受体的分布、雌激素对胎儿和新生儿睾丸发育以及对精子发生的影响这几个方面进行了综述.     

性激素受体在子午岭黑山羊隐睾及正常睾丸中的分布比较

旨在研究性激素受体[雄激素受体（androgen receptor,AR）、雌激素受体（estrogen receptor,ER）]在子午岭黑山羊正常睾丸与隐睾组织中的分布与隐睾症的关系,应用免疫组织化学及免疫荧光技术方法结合形态计量学统计软件,比较了正常睾丸与隐睾的组织化学特点。特殊染色结果显示：与正常组相比,隐睾组间质组织疏松,管腔面积明显减小,糖原含量明显减少,胶原纤维及网状纤维含量增多。免疫组化及免疫荧光结果显示：1） AR在正常睾丸组Leydig细胞呈高密度强阳性表达,在各级生精细胞呈中等强度阳性表达,隐睾组中Leydig细胞及各级生精细胞表达明显减弱,且在精原细胞偶见表达;2） ER在正常睾丸Leydig细胞呈高密度强阳性表达,管周肌样细胞呈中等强度阳性表达,Sertoli细胞偶见表达,各级生精细胞无表达;3） ER在隐睾Leydig细胞、初级精母细胞、精原细胞和Sertoli细胞中均呈中等强度阳性表达;4）统计结果显示,隐睾组AR的平均光密度较正常组显著降低（P<0.05）,而ER的平均光密度则显著高于正常组（P<0.01）,且隐睾组AR与ER表达量比值基本接近1：1。子午岭黑山羊隐睾组织胶原纤维和网状纤维分布较正常睾丸多,生精小管基膜主要成分以中性糖蛋白为主,酸性糖蛋白含量明显降低影响精子的正常形成;隐睾组织精原细胞及Sertoli细胞ER与AR表达失常尤为明显,可为哺乳动物隐睾的相关研究提供一定参考。     

雌激素受体与动物性别发育关系的研究进展

近年来,"鸡体细胞自主性性别决定机制"的提出使得鸡性别决定的研究倍受关注.在鸡中,雌激素虽然可以在一定程度上影响性表型,但起到的作用却远不如哺乳动物中那么显著,这可能与雌激素发挥作用的载体--雌激素受体(ER)在不同物种中的功能差异有关.因此,本文就雌激素受体的结构、作用机制、在不同物种中的表达规律及其在性别发育上的作...     

Immunohistochemical expression of estrogen receptor beta in normal and tumoral canine mammary glands

To date, two isoforms of estrogen receptors (ER) have been identified, cloned, and characterized from several species, estrogen receptor-alpha (ERalpha) and estrogen receptor-beta (ERbeta). Although the presence of ERalpha has been demonstrated in normal and tumoral canine mammary tissues, the issue of ERbeta expression has not been addressed in the dog. In this study, we have analyzed the expression of ERbeta in formalin-fixed, paraffin-embedded tissue samples of nonaltered mammary gland, 30 malignant (six complex carcinoma, 12 simple carcinoma, three carcinosarcoma, and nine carcinoma or sarcoma in benign tumor), and five benign (one fibroadenoma, one complex papilloma, one complex adenoma, and two benign mixed tumors) mammary tumors of the dog by using a polyclonal ERbeta antibody and the avidin-biotin-peroxidase complex immunohistochemical technique. Our results show that high numbers of normal ductal and acinar epithelium and approximately one third of canine mammary tumors express ERbeta. This expression was higher in benign than in malignant tumors. Furthermore, expression was higher in complex and mixed histologic subtypes of malignant tumors when compared with simple subtypes.     

Changes in estrogen receptor expression in the chick thymus during late embryonic development

In chickens, although estrogen receptors (ER) are reported to be associated with the immunological processes, detailed information about the differences in ER expression in the tissues related to the development of lymphocytes is not fully known, especially during the developmental stage. To learn more about this immunological relationship, we used semi‐quantitative polymerase chain reaction method to detect the ER expression levels in the thymus tissues of chicks during the developmental stage. Furthermore, ER‐expressing cells were detected by immunohistochemistry. The results of this study show that the expression level of ER increased on embryonic day 16 and decreased on day 20. Furthermore, ER expression was significantly higher in male than in female chickens at day 16. The increased expression on day 16 and decreased level on day 20 were also reproduced in the incidence of immunoreactive cells, although there was a 1‐day delay in the elevated incidence of the cells. This study revealed the changes in ER expression and the incidence of ER‐positive cells in the thymus of chickens during the developmental stage.     

Expression of estrogen receptor alpha and beta in the uterus and vagina of immature rats treated with 17-ethinyl estradiol

The action of estrogen on target organs has been actively studied with the discovery of estrogen receptor (ER) beta. This study was carried out to examine the expression of ERalpha and ERbeta in the uterus and the vagina of immature Sprague-Dawley rats treated with 17-ethinyl estradiol (EE). Twenty days old rats were subcutaneously treated with EE at the doses of 0 (vehicle control), 0.03, 0.3, 1.0, 3.0, and 10.0 microg/kg/day for three consecutive days. The treatment of EE at the doses of 0.3, 1.0, 3.0 and 10.0 microg/kg/day significantly increased the weights of the uterus and vagina of rats (p<0.01) and retained fluid in the uterus of rats. At the high doses of 3.0 and 10.0 microg/kg/day, the treatment of EE caused an increase in the uterine height, hypertrophy, and a decrease in the expression of ERalpha and ERbeta in the uterine luminal and glandular epithelium. The treatment of EE at the doses of 3.0 and 10.0 microg/kg/day also caused cornification and a decrease in the expression of ERalpha and ERbeta in the vaginal epithelium. These results suggest that the EE treatment decrease the expression of ERalpha and ERbeta in the uterus and vagina of immature rats and that may be associated with the morphological changes such as increase in the uterine height, hypertrophy of the uterine epithelium, and cornification of the vagina.     

Effects of estrogen on estrogen receptor expression in the bursal cells of chick embryos and steroidogenic enzymes gene expression in the bursa: Relevance of estrogen receptor and estrogen synthesis in the bursa of chick embryos

Estrogen has been reported to act on B cell genesis in the bursa of Fabricius of chick embryos. In this study, we attempted to demonstrate the hypothesis that B cell genesis is controlled by estrogen receptor (ER) in the bursal cells and steroidogenic enzymes synthesized in the bursa. We previously reported the presence of estrogen receptor α (ERα) in the bursa during the late stage of embryogenesis and an increase in the expression of ERα messenger RNA (mRNA) between the 13th day and 16th day. The number of ER-positive cells was maximal on the 16th day. In the present study, ER-positive cells in the bursa during the late stage of embryogenesis increased 4 h after β-estradiol treatment on the 14th to 18th day. The concentration of β-estradiol in the embryonic bursa increased. These results suggest that this stage of embryogenesis is critical in B cell development in the bursa in connection with the effect of estrogen treatment. Our findings also showed that the mRNA expression of five steroidogenic enzymes occurred in the bursa of chick embryos. These results suggest that estrogen is synthesized in the embryonic bursa and estrogen acts on the bursal cells in a paracrine fashion.     

Expression of estrogen receptor alpha (ERalpha) and estrogen receptor beta (ERbeta) in the ovarian follicles and corpora lutea of pregnant swine

The objective of the study was to demonstrate the presence of estrogen receptor alpha (ERalpha) and beta (ERbeta) protein and corresponding mRNA in porcine ovarian follicles and corpora lutea obtained on day 10, 18, 32, 50, 71 and 90 post coitum (p.c.) using immunohistochemistry, Western blot, and RT-PCR analysis. Immunohistochemistry showed that ERalpha protein was located in the granulosa cells of ovarian follicles and the strongest immunoreaction was observed on days 32 and 50 p.c. The ERbeta protein was found mainly in theca cells of follicles as well as in luteal cells. The most intense immunoreaction was observed on day 18 p.c. within theca cells, while in the corpus luteum (CL) the intensity of ERbeta staining gradually increased and remained elevated at mid and late pregnancy. In CL by day 50 p.c. immunoreaction for ERbeta was present only in small luteal cells, but starting from day 71 to 90 p.c. it was observed in both small and large luteal cells. Western blot analysis was performed and validated data obtained from immunohistochemistry. RT-PCR results indicated that ERalpha mRNA was expressed only in ovarian follicles of the pregnant swine, while that of ERbeta in both follicles and CL. The results suggest an autocrine/paracrine role of estrogens acting via both ERalpha and ERbeta in the regulation of the ovarian function during pregnancy and for the process of successful reproduction.     

成年公猪睾丸及附睾中促黄体素受体mRNA表达的研究

试验采用FQ-PCR技术进行了成年公猪睾丸及附睾中促黄体素受体mRNA表达的研究,结果表明:促黄体素受体mRNA不仅在睾丸内表达,在附睾中也有表达;睾丸、附睾中促黄体素受体mRNA的表达量分别为(1.42±0.08)×109 copies/mL和(6.54±0.21)×109copies/mL,睾丸两组织间促黄体素受体mRNA表达量无明显差异(P>0.05).     

不同光照时长对鸡睾丸褪黑素受体及合成酶表达的影响

旨在研究不同光照时长对鸡睾丸组织中褪黑素受体及合成酶表达规律的影响。选用20周龄体质量相近的AA(爱拔益加)种公鸡共120只,随机分为3组,分别使用8.0,11.5和16.0h光照处理,饲喂4周,饲喂结束后颈动脉放血致死,采取睾丸。采用实时荧光定量PCR和Western blot方法检测鸡睾丸组织中褪黑素受体及合成酶mRNA和蛋白表达量。结果表明,鸡睾丸组织中褪黑素受体MEL-1A、MEL-1B和MEL-1C,褪黑素合成酶AANAT的mR-NA及蛋白表达量随光照时长增加而显著降低(P0.01);但合成酶ASMT表达量无显著变化(P0.05)。综上,褪黑素受体MEL-1A、MEL-1B和MEL-1C及合成酶AANAT的表达与光照时长呈负相关。     

The expression and localization of inhibin isotypes in mouse testis during postnatal development

Inhibin, which is important for normal gonadal function, acts on the pituitary gonadotropins to suppress follicle-stimulating hormone (FSH) secretion. The level and cellular localization of the inhibin isotypes, α, β_A and β_B, in the testis of mice were examined during postnatal development in order to determine if inhibin expression is related to testicular maturation. Mouse testes were sampled on postnatal days (PNDs) 1, 3, 6, 18, 48 and 120, and analyzed by Western blotting and immunofluorescence. Western blot analysis showed very low levels of inhibin α, β_A and β_B expression in the testes at days 1 to 6 after birth. The levels then increased gradually from PND 18 to 48-120, and there were significant peaks at PND 48. Inhibin α, β_A and β_B were detected in testicular cells during postnatal development using immunohistochemistry. The immunoreactivity of inhibin α was rarely observed in testicular cells during PND 1 to 6, or in the cytoplasmic process of Sertoli cells surrounding the germ cells and interstitial cells during PND 18 to 120. Inhibin β_A and β_B immunoreactivity was rarely observed in the testis from PND 1 to 6. On the other hand, it was observed in some spermatogonial cells, as well as in the interstitial space between PND 48 and PND 120. We conclude that the expression of inhibin isotypes increases progressively in the testis of mice with increasing postnatal age, suggesting that inhibin is associated with a negative feedback signal for FSH in testicular maturation.     

Evaluation of progesterone and estrogen receptor expression in 15 meningiomas of dogs and cats

OBJECTIVE: To evaluate progesterone and estrogen receptor expression in meningiomas of the CNS in dogs and cats. ANIMAL: 8 dogs (1 of which was treated with gestrinone) and 5 cats with intracranial meningiomas and 2 dogs with spinal cord meningiomas; tissue samples were also obtained from 1 clinically normal dog and 1 clinically normal cat. PROCEDURE: Meningioma tissue was obtained during surgery or at necropsy; samples were processed for histologic classification and immunohistochemical evaluation of the proportion of tumor cells with progesterone and estrogen receptors. Correlation among receptor expression, tumor grade, and histologic subtypes was determined. RESULT: Several histologic subtypes of intracranial meningiomas were detected among tissue samples. In the cats, all intracranial meningiomas were benign. Progesterone receptor immunoreactivity was detected in 14 of 15 meningiomas. Progesterone receptor expression was identified in > 80% of cells in 8 intracranial meningiomas (4 dogs and 4 cats) and 2 spinal cord meningiomas. In samples of malignant transitional and granular cell meningiomas in dogs, progesterone receptors were detected in 32 and 4.8% of cells respectively. In 1 cat, 38% of tumor cells had progesterone receptors. In a dog treated with gestrinone, no progesterone receptors were detected in the intracranial meningioma. Estrogen receptors were only detected in the tumor of 1 dog. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate a high proportion of progesterone receptors in cells of meningiomas of the CNS in dogs and cats. Antiprogesterone treatment may have a role in the treatment of unresectable or recurrent meningiomas in dogs and cats.