TRANSIENT EXPRESSION OF PROGESTERONE-RECEPTOR MESSENGER-RNA IN OVARIAN GRANULOSA-CELLS AFTER THE PREOVULATORY LUTEINIZING-HORMONE SURGE
TRANSIENT EXPRESSION OF PROGESTERONE-RECEPTOR MESSENGER-RNA IN OVARIAN GRANULOSA-CELLS AFTER THE PREOVULATORY LUTEINIZING-HORMONE SURGE
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DOI：
10.1210/mend-5-7-967
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发表时间：
1991-07-01
影响因子：
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通讯作者：
MAYO, KE
中科院分区：
文献类型：
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作者：
PARK, OK;MAYO, KE
卵巢类固醇孕酮通过调节靶组织中特定基因的表达而影响生殖生理。为了解决卵巢本身是否是孕酮作用的靶组织的问题，我们研究了孕激素受体(PR)mRNA在大鼠卵巢中的定位和调节。我们利用聚合酶链式反应从子宫中克隆了大鼠PR的类固醇结合结构域，并以此为探针从大鼠胎盘cDNA文库中分离了一个更大的cDNA.我们用RNA过滤杂交、定量逆转录聚合酶链式反应扩增和原位杂交检测了大鼠卵巢中PR基因的表达。PR基因的表达最初是在一个未成熟的动物模型中进行的；23日龄的大鼠接受了PMSG或PMSG+hCG的治疗。我们在对照组和PMSG处理组的卵巢中几乎没有发现PR基因的表达，而在PMSG和hCG处理的动物的卵巢中，大卵泡的颗粒细胞中PRmRNA的表达水平很高。其他类型的细胞，包括膜细胞和间质细胞，不表达可检测到的PR mRNA水平。人绒毛膜促性腺激素(HCG)注射后5h，未成熟卵巢中PR基因表达增加20倍以上，12h后表达下调至接近基础水平。在随后的一系列实验中，我们检测了成年大鼠发情周期中PR基因的表达。PR基因的表达是短暂的，并与发情前黄体生成素高峰密切相关。在发情周期中，PR mRNA定位于成熟卵泡的颗粒细胞。在用戊巴比妥阻断排卵前黄体生成素峰的周期动物中，未观察到发情后期PRmRNA的诱导。PR基因在大鼠卵巢中的这种短暂的、激素调节的和细胞特异性的表达强烈地表明，在大鼠生殖周期中，孕酮在卵巢内具有重要的功能。
The ovarian steroid progesterone affects reproductive physiology by regulating the expression of specific genes in target tissues. In an attempt to address the question of whether the ovary itself is a target tissue for progesterone action, we have examined the localization and regulation of progesterone receptor (PR) mRNA in the rat ovary. We used the polymerase chain reaction to clone the steroid-binding domain of the rat PR from uterine cDNA and used this as a probe to isolate a larger cDNA from a rat placental cDNA library. We used RNA filter hybridization, a quantitative reverse transcription-polymerase chain reaction amplification assay, and in situ hybridization to detect PR mRNA in the rat ovary. Expression of the PR gene was initially studied in an immature animal model; 23-day-old rats were treated with either PMSG or PMSG followed by hCG. We found little or no PR mRNA in the ovaries of control or PMSG-treated animals; however, the mRNA was highly expressed in the granulosa cells of large follicles in the ovaries of animals treated with PMSG followed by hCG. Other cell types, including thecal and interstitial cells, did not express detectable levels of PR mRNA. The PR mRNA was induced more than 20-fold in the immature ovary 5 h after hCG administration and was down-regulated to near-basal levels by 12 h after hCG administration. In a subsequent series of experiments, we examined PR gene expression in adult rats during the estrous cycle. The expression of PR mRNA was transient and was tightly coupled to the preovulatory LH surge on proestrous evening. PR mRNA was localized to the granulosa cells of mature ovarian follicles during the estrous cycle. In cycling animals treated with pentobarbital to block the preovulatory LH surge, no induction of PR mRNA on proestrous evening was observed. This transient, hormonally regulated, and cell-specific expression of the PR gene in the rat ovary strongly suggests an important intraovarian function for progesterone during the rat reproductive cycle.