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发情周期卵泡期和黄体期印度水牛输卵管中雌激素受体α(ERα)的免疫组织化学定位

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

作者信息

Pathak Devendra, Bansal Neelam, Singh Opinder, Gupta Kuldip, Ghuman S P S

机构信息

Department of Veterinary Anatomy, College of Veterinary Science, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab, 140004, India.

Department of Veterinary Pathology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab, India.

出版信息

Trop Anim Health Prod. 2019 Jul;51(6):1601-1609. doi: 10.1007/s11250-019-01852-y. Epub 2019 Mar 2.

Abstract

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.

摘要

采用免疫组织化学方法,研究了水牛发情周期卵泡期和黄体期输卵管不同节段雌激素受体α(ERα)的定位和分布。在评估卵巢大体形态后,从屠宰场收集12头水牛(发情周期卵泡期和黄体期各6头)输卵管不同节段的组织样本。此外,在屠宰前采集动物血液样本,以估计雌激素和孕激素水平。使用一步法聚合物HRPO染色系统,通过免疫组织化学技术确定雌激素受体的组织分布。雌激素受体定位于上皮层、黏膜下层、肌层和浆膜层。在上皮层观察到最大定位,其中纤毛细胞和分泌细胞类型的ERα均呈阳性。发情周期卵泡期和黄体期阳性细胞百分比有所不同。输卵管腺的衬里上皮对ERα也呈强阳性。当用抗体稀释剂或缓冲液代替一抗时,在输卵管的任何一层均未观察到免疫染色,作为阴性对照。数据显示,在输卵管壶腹部区域观察到最高免疫阳性细胞,而在子宫输卵管交界处这些细胞最少(p<0.05)。与发情周期黄体期相比,发情周期卵泡期漏斗部、壶腹部和峡部显示出更高百分比的ERα阳性细胞(p<0.05)。子宫输卵管交界处发情周期两个阶段的阳性细胞百分比无显著差异。用抗ERα抗体进行免疫金标记在亚细胞水平证实了免疫组织化学研究的结果。卵泡期较高的表达与雌激素水平直接相关。

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