Knapczyk Katarzyna, Duda Malgorzata, Durlej Malgorzata, Galas Jerzy, Koziorowski Marek, Slomczynska Maria
Department of Endocrinology and Tissue Culture, Institute of Zoology, Jagiellonian University, Ingardena 6, Krakow, Poland.
Domest Anim Endocrinol. 2008 Aug;35(2):170-9. doi: 10.1016/j.domaniend.2008.05.001. Epub 2008 Jun 6.
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.
本研究的目的是利用免疫组织化学、蛋白质印迹法和逆转录-聚合酶链反应(RT-PCR)分析,证明在交配后第10、18、32、50、71和90天获得的猪卵巢卵泡和黄体中存在雌激素受体α(ERα)和β(ERβ)蛋白及相应的mRNA。免疫组织化学显示,ERα蛋白位于卵巢卵泡的颗粒细胞中,在交配后第32天和50天观察到最强的免疫反应。ERβ蛋白主要存在于卵泡的膜细胞以及黄体细胞中。在交配后第18天,在膜细胞中观察到最强烈的免疫反应,而在黄体中,ERβ染色强度逐渐增加,并在妊娠中期和晚期保持升高。在交配后第50天的黄体中,ERβ的免疫反应仅存在于小黄体细胞中,但从交配后第71天到90天,在小黄体细胞和大黄体细胞中均观察到。进行了蛋白质印迹分析,并验证了从免疫组织化学获得的数据。RT-PCR结果表明,ERα mRNA仅在妊娠母猪的卵巢卵泡中表达,而ERβ的mRNA在卵泡和黄体中均有表达。结果表明,雌激素通过ERα和ERβ发挥自分泌/旁分泌作用,在妊娠期间调节卵巢功能以及成功繁殖过程中发挥作用。