G protein-coupled receptor 30 expression is required for estrogen stimulation of primordial follicle formation in the hamster ovary
G protein-coupled receptor 30 expression is required for estrogen stimulation of primordial follicle formation in the hamster ovary
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DOI：
10.1210/en.2008-0441
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发表时间：
2008-09-01
期刊：
影响因子：
4.8
通讯作者：
Roy, Shyamal K.
中科院分区：
文献类型：
--
作者：
Wang, Cheng;Prossnitz, Eric R.;Roy, Shyamal K.
雌二醇17β（E-2）在原始卵泡的形成和发育中发挥着重要作用，但其机制仍不清楚。 G 蛋白偶联受体 30 (GPR30) 可以在多种细胞中介导快速且不依赖于转录的 E2 信号传导。本研究的目的是检测 GPR30 在新生仓鼠卵巢中是否表达以及它是否可以在原始卵泡形成过程中介导雌激素作用。 GPR30 mRNA 水平从妊娠第 13 天 (E13) 到产后第二天 (P2) 下降，然后从 P3 到 P6 稳定增加。与 mRNA 水平的变化一致，GPR30 蛋白表达从 E13 到 P2 下降，随后在 P7（仓鼠卵巢中首次出现原始卵泡的前一天）显着增加。 GPR30 在卵母细胞和体细胞中均表达，但在卵母细胞中表达较低。 GPR30蛋白主要位于核周内质网，这也是E-2-BSA-FITC（E2-BSA-异硫氰酸荧光素）结合的位点。 E-2 或 E-2-BSA 在体外增加新生仓鼠卵巢细胞的细胞内钙。体外暴露于 GPR30 小干扰 RNA 显着降低了培养的仓鼠卵巢中的 GPR30 mRNA 和蛋白质水平，减弱了 E-BSA 与培养的 P6 卵巢细胞的结合，并显着抑制了雌激素刺激的原始卵泡形成。这些结果表明，膜雌激素受体 GPR30 在围产期发育过程中在卵巢中表达，并介导 E-2 对原始卵泡形成的作用。
Estradiol-17 beta (E-2) plays an important role in the formation and development of primordial follicles, but the mechanisms remain unclear. G protein-coupled receptor 30 (GPR30) can mediate a rapid and transcription-independent E2 signaling in various cells. The objectives of this study were to examine whether GPR30 was expressed in the neonatal hamster ovary and whether it could mediate estrogen action during the formation of primordial follicles. GPR30 mRNA levels decreased from the 13th day of gestation (E13) through the second day of postnatal (P2) life, followed by steady increases from P3 through P6. Consistent with the changes in mRNA levels, GPR30 protein expression decreased from E13 to P2 followed by a significant increase by P7, the day before the first appearance of primordial follicles in the hamster ovary. GPR30 was expressed both in the oocytes and somatic cells, although the expression in the oocytes was low. GPR30 protein was located primarily in the perinuclear endoplasmic reticulum, which was also the site of E-2-BSA-FITC (E2-BSA-fluorescein isothiocyanate) binding. E-2 or E-2-BSA increased intracellular calcium in neonatal hamster ovary cells in vitro. Exposure to GPR30 small interfering RNA in vitro significantly reduced GPR30 mRNA and protein levels in cultured hamster ovaries, attenuated E-BSA binding to cultured P6 ovarian cells, and markedly suppressed estrogen-stimulated primordial follicle formation. These results suggest that a membrane estrogen receptor, GPR30, is expressed in the ovary during perinatal development and mediates E-2 action on primordial follicle formation.