Chaffin C L, Heimler I, Rawlins R G, Wimpee B A, Sommer C, Hutz R J
Department of Biological Sciences, University of Wisconsin-Milwaukee, 3209 N. Maryland Ave., 53211, Milwaukee, WI.
Endocrine. 1996 Dec;5(3):315-21. doi: 10.1007/BF02739065.
We have previously shown by immunocytochemistry and autoradiography the presence of estrogen receptors (ER) in rhesus monkey ovary. Intense chromogen staining showed specific binding for ER in nuclei of germinal epithelium and granulosa cells of antral follicles; and radiolabeled ligand bound specifically to functional corpora lutea (CL). Although it is accepted that the germinal epithelium of the primate ovary contains ER, some controversy still persists regarding the intraovarian localization of this molecule. In addition, no data exist that localize the aromatic hydrocarbon (dioxin) receptor (AHR), which is known to modulate ER, to the primate ovary. In the present study, we show the presence of ER using Western blot analysis, and ER capable of binding DNA within intraovarian compartments in two species of the genusMacaca (rhesus macaque,Macaca mulatta and stumptail macaque,Macaca arctoides); extend these findings to human ovarian granulosa cells (GC) using Western blot, reverse transcription-polymerase chain reaction (RT-PCR), and gel mobility-shift analysis; and localize the AHR to intraovarian compartments of the macaque ovary by Western blots and gel-shift assays. These experiments strongly suggest that estrogens can exert effects on follicle development directly at the ovary, and provide the first direct evidence that AHR-mediated toxicity may be manifested at the ovary to induce possible antifertility effects.
我们之前通过免疫细胞化学和放射自显影法证明了恒河猴卵巢中存在雌激素受体(ER)。强烈的显色剂染色显示,在窦状卵泡的生发上皮细胞核和颗粒细胞中,ER有特异性结合;放射性标记配体与功能性黄体(CL)特异性结合。尽管人们公认灵长类动物卵巢的生发上皮含有ER,但关于该分子在卵巢内的定位仍存在一些争议。此外,尚无数据表明已知可调节ER的芳烃(二噁英)受体(AHR)在灵长类动物卵巢中的定位。在本研究中,我们通过蛋白质印迹分析证明了ER的存在,以及在猕猴属的两个物种(恒河猴,猕猴和短尾猕猴,熊猴)的卵巢内隔室中能够结合DNA的ER;使用蛋白质印迹、逆转录聚合酶链反应(RT-PCR)和凝胶迁移率变动分析将这些发现扩展到人类卵巢颗粒细胞(GC);并通过蛋白质印迹和凝胶迁移率变动分析将AHR定位到猕猴卵巢的卵巢内隔室。这些实验强烈表明,雌激素可直接在卵巢对卵泡发育产生影响,并提供了首个直接证据,证明AHR介导的毒性可能在卵巢表现出来,从而诱导可能的抗生育作用。