Hormonal regulation of follicle-stimulating hormone receptor messenger ribonucleic acid levels in cultured rat granulosa cells.
Hormonal regulation of follicle-stimulating hormone receptor messenger ribonucleic acid levels in cultured rat granulosa cells.
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DOI：
10.1210/endo.130.3.1311235
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发表时间：
1992-03
期刊：
影响因子：
4.8
通讯作者：
J. Tilly;P. Lapolt;A. Hsueh
中科院分区：
文献类型：
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作者：
J. Tilly;P. Lapolt;A. Hsueh
卵巢颗粒细胞的成熟依赖于垂体促性腺激素FSH，其作用是通过特定的质膜受体介导的。为了在mRNA水平上研究卵巢FSH受体表达的调控，我们使用特异性cRNA探针来评估培养颗粒细胞中FSH受体转录本的变化。未成熟雌激素处理大鼠的颗粒细胞含有两种主要的FSH受体mRNA转录物（7.0和2.5千碱基），在2天的培养期间，其水平呈时间相关下降。然而，在培养基中加入FSH （30 ng/ml）可以阻止FSH受体mRNA水平的下降。与对照组相比，用FSH （1-100 ng/ml）处理颗粒细胞48小时后，FSH受体mRNA水平呈剂量依赖性增加（ED50, 4.5 ng/ml），当FSH浓度为30 ng/ml时，最大增幅为5.9 +/- 0.7倍。腺苷环化酶激活剂forskolin（0.1-30微米）可以模拟FSH的刺激作用，提示cAMP参与FSH受体基因转录和/或mRNA稳定性。颗粒细胞与表皮生长因子（EGF, 0.3-10 ng/ml）、碱性成纤维细胞生长因子（bFGF, 1-30 ng/ml）或胰岛素样生长因子-1 （igf -1, 1-30 ng/ml）孵育48小时，不影响FSH受体mRNA水平，而最高剂量的EGF和bFGF，而不是igf -1，完全抑制FSH （30 ng/ml）对其自身受体mRNA水平的刺激作用。同样，GnRH （10-1000 nM）以剂量依赖性的方式减弱FSH对其受体mRNA水平的作用（ID50, 8 nM）。通过与GnRH拮抗剂（[Ac-D-Phe1,D-pCl-Phe2,D-Trp3,6]GnRH; 100 nM）共处理，GnRH （100 nM）的抑制作用被逆转，表明GnRH的作用是通过特异性GnRH受体介导的。这些数据表明，用FSH处理颗粒细胞增加了两种FSH受体mRNA转录物的水平。然而，这种可能导致FSH作用放大的正反馈系统受到EGF、bFGF和GnRH抑制作用的严格调控。因此，利用培养的大鼠颗粒细胞为分析卵巢中激素对FSH受体基因表达的调节提供了一个模型系统。
The maturation of ovarian granulosa cells is dependent upon the pituitary gonadotropin FSH, the actions of which are mediated via specific plasma membrane receptors. To study the regulation of ovarian FSH receptor expression at the mRNA level, we used a specific cRNA probe to evaluate changes in FSH receptor transcripts in cultured granulosa cells. Granulosa cells obtained from immature estrogen-treated rats contained two predominant FSH receptor mRNA transcripts (7.0 and 2.5 kilobases), the levels of which declined in a time-related manner during a 2-day culture period. However, inclusion of FSH (30 ng/ml) in the culture medium prevented the decline in FSH receptor mRNA levels. Compared to controls, treatment of granulosa cells for 48 h with FSH (1-100 ng/ml) increased FSH receptor mRNA levels in a dose-dependent manner (ED50, 4.5 ng/ml), with a maximal 5.9 +/- 0.7-fold increase observed in response to 30 ng/ml FSH. The stimulatory actions of FSH were mimicked by the adenyl cyclase activator forskolin (0.1-30 microM), suggesting the involvement of cAMP in FSH receptor gene transcription and/or mRNA stability. Incubation of granulosa cells for 48 h with epidermal growth factor (EGF; 0.3-10 ng/ml), basic fibroblast growth factor (bFGF; 1-30 ng/ml), or insulin-like growth factor-I (IGF-I; 1-30 ng/ml) did not affect basal FSH receptor mRNA levels, whereas the highest doses of EGF and bFGF, but not IGF-I, completely suppressed the stimulatory effects of FSH (30 ng/ml) on its own receptor mRNA levels. Similarly, GnRH (10-1000 nM) attenuated the actions of FSH on its receptor mRNA levels in a dose-dependent manner (ID50, 8 nM). The inhibitory effects of GnRH (100 nM) were reversed by cotreatment with a GnRH antagonist ([Ac-D-Phe1,D-pCl-Phe2,D-Trp3,6]GnRH; 100 nM), indicating that the actions of GnRH are mediated via specific GnRH receptors. These data indicate that treatment of granulosa cells with FSH increases the levels of two FSH receptor mRNA transcripts. However, this positive feedback system, which may lead to an amplification of FSH action, is tightly regulated by the inhibitory actions of EGF, bFGF, and GnRH. Thus, the use of cultured rat granulosa cells provides a model system to analyze the hormonal regulation of FSH receptor gene expression in the ovary.