Agarwal P, Peluso J J, White B A
Department of Anatomy, University of Connecticut Health Center, Farmington 06030, USA.
Biol Reprod. 1996 Dec;55(6):1271-5. doi: 10.1095/biolreprod55.6.1271.
The expression of steroidogenic factor 1 (SF-1), cytochrome P450 aromatase (P450arom), and cytochrome P450 cholesterol side-chain cleavage (P450scc) was examined during the periovulatory period. Immature rats were injected with eCG to induce development of ovarian follicles to the preovulatory stage. At 48 h after the eCG injection, the LH surge was simulated by an injection of an ovulatory dose of hCG, and RNA was isolated at 0, 2, 4, 6, 8, and 24 h after hCG injection. The mRNA levels for SF-1, P450arom, and P450scc were relatively high in total ovarian RNA samples from eCG-treated rats. Levels of SF-1 and P450arom mRNA decreased within 2 h after injection of hCG. The SF-1 mRNA levels gradually increased from 4 to 24 h. Aromatase levels remained undetectable until 24 h after hCG. P450scc mRNA levels increased throughout the 24-h period after hCG. Levels of SF-1 and P450arom, but not P450scc, mRNA were also reduced in RNA samples from isolated granulosa cells at 4 h after hCG treatment relative to those from eCG-treated rats. In situ hybridization analysis also revealed that hCG uniformly suppressed SF-1 mRNA levels an all granulosa cells compared to those of eCG-treated controls. The relationship of SF-1 expression to immediate/early gene expression and cell cycle traverse was also examined. C-myc mRNA levels were induced by up to 10-fold at 4 h after hCG injection. Similarly, DNA synthesis, as measured by the percentage of granulosa cells that incorporated 5'-bromodeoxyuridine, was increased from 16 +/- 4% in eCG-treated rats to 61 +/- 7% at 4 h after hCG treatment (p < 0.05). This study provides the novel finding that SF-1 expression is transiently repressed to very low levels in response to the LH surge. Further, these studies suggest that granulosa cells traverse the cell cycle before becoming terminally differentiated luteal cells.
在排卵期前后,对类固醇生成因子1(SF-1)、细胞色素P450芳香化酶(P450arom)和细胞色素P450胆固醇侧链裂解酶(P450scc)的表达进行了检测。给未成熟大鼠注射eCG以诱导卵巢卵泡发育至排卵前期。在注射eCG后48小时,通过注射排卵剂量的hCG模拟LH峰,并在注射hCG后的0、2、4、6、8和24小时分离RNA。在经eCG处理的大鼠的总卵巢RNA样本中,SF-1、P450arom和P450scc的mRNA水平相对较高。注射hCG后2小时内,SF-1和P450arom mRNA水平下降。SF-1 mRNA水平在4至24小时逐渐升高。直到hCG注射后24小时,芳香化酶水平仍无法检测到。hCG处理后4小时,分离的颗粒细胞RNA样本中SF-1和P450arom mRNA水平相对于经eCG处理的大鼠也降低,但P450scc mRNA水平未降低。原位杂交分析还显示,与经eCG处理的对照组相比,hCG使所有颗粒细胞中的SF-1 mRNA水平均一性降低。还研究了SF-1表达与即刻/早期基因表达和细胞周期进程的关系。hCG注射后4小时,C-myc mRNA水平诱导增加高达10倍。同样,通过掺入5'-溴脱氧尿苷的颗粒细胞百分比测量的DNA合成,从经eCG处理的大鼠中的16±4%增加到hCG处理后4小时的61±7%(p<0.05)。本研究提供了一个新发现,即SF-1表达在LH峰的作用下被短暂抑制至极低水平。此外,这些研究表明颗粒细胞在成为终末分化的黄体细胞之前经历细胞周期。