Yang Peixin, Kriatchko Alexie, Roy Shyamal K
Department of Obstetrics and Gynecology, University of Nebraska Medical Center, Omaha, Nebraska 68198-4515, USA.
Endocrinology. 2002 Jun;143(6):2385-98. doi: 10.1210/endo.143.6.8858.
Spatiotemporal expression patterns of ER-alpha and ER-beta protein and mRNA in hamster ovarian cells during the estrous cycle and following hypophysectomy and selective hormone replacement were evaluated by immunofluorescence, immunoblotting and in situ hybridization analyses. Whereas ER-beta mRNA and protein expression predominated in granulosa cells and ER-alpha expression was in interstitial and thecal cells, overlap in receptor subtype expression across cell types was evident. Both ER subtypes were present from primordial follicle stage onward. ER-alpha mRNA levels and immunoreactivity started increasing from D3:0900 h in interstitial and granulosa cells and peaked on the proestrous (D4:0900 h). Regionalized higher expression of ER-alpha in granulosa cells in and around the forming antrum was evident. Surface epithelial cells were also positive. ER-beta mRNA and protein expression increased markedly in granulosa and interstitial cells on D2:0900 h, reached a peak on D3:0900 h, and then declined sharply on D4:0900 h. No change in ER expression occurred following the preovulatory gonadotropin surge. Whereas FSH or human CG stimulated ER-alpha mRNA and protein expression in hypophysectomized hamsters, only FSH could stimulate ER-beta mRNA and protein, and the effect was significantly attenuated by human CG. ER expression was stimulated by estrogen, but progesterone strongly inhibited estrogen action. These results indicate that ER expression is cell type specific to the larger extent and is critically regulated by reproductive hormones.
通过免疫荧光、免疫印迹和原位杂交分析,评估了仓鼠卵巢细胞在发情周期、垂体切除术后以及选择性激素替代后,雌激素受体α(ER-α)和雌激素受体β(ER-β)蛋白及mRNA的时空表达模式。虽然ER-β mRNA和蛋白表达在颗粒细胞中占主导,而ER-α表达存在于间质细胞和卵泡膜细胞中,但不同细胞类型间受体亚型表达的重叠是明显的。两种ER亚型从原始卵泡阶段就已存在。ER-α mRNA水平和免疫反应性从第3天09:00开始在间质细胞和颗粒细胞中增加,并在动情前期(第4天09:00)达到峰值。在形成的卵泡腔及其周围的颗粒细胞中,ER-α有明显的区域化高表达。表面上皮细胞也呈阳性。ER-β mRNA和蛋白表达在第2天09:00在颗粒细胞和间质细胞中显著增加,在第3天09:00达到峰值,然后在第4天09:00急剧下降。排卵前促性腺激素激增后,ER表达没有变化。虽然促卵泡激素(FSH)或人绒毛膜促性腺激素(hCG)刺激垂体切除的仓鼠中ER-α mRNA和蛋白表达,但只有FSH能刺激ER-β mRNA和蛋白,且该作用被hCG显著减弱。雌激素刺激ER表达,但孕酮强烈抑制雌激素作用。这些结果表明,ER表达在很大程度上具有细胞类型特异性,并受到生殖激素的严格调控。