Involvement of progesterone in gonadotrophin-induced pituitary adenylate cyclase-activating polypeptide gene expression in pre-ovulatory follicles of rat ovary.
Involvement of progesterone in gonadotrophin-induced pituitary adenylate cyclase-activating polypeptide gene expression in pre-ovulatory follicles of rat ovary.
复制标题
黄体酮参与促性腺激素诱导的大鼠卵巢排卵前卵泡中垂体腺苷酸环化酶激活多肽基因的表达。
作者：
Jae;Wan;Li Wang;Hyun Jeong Park;Jin Lee;Jeong;Hyuk;A. Tsafriri;S. Chun
本研究旨在探讨黄体酮是否参与促性腺激素诱导的垂体腺苷酸环化酶激活多肽（Pacap）基因在大鼠卵巢中的表达。Northern blot分析显示，孕母马血清促性腺激素（PMSG）引发的未成熟大鼠在HCG前1小时用孕激素拮抗剂RU486或3β -羟基类固醇脱氢酶epostane抑制剂处理，导致HCG诱导的Pacap基因表达呈剂量依赖性抑制。原位杂交表明，RU486处理后卵巢颗粒细胞中表达Pacap mRNA的排卵前卵泡数量大大减少。此外，RU486或epostane与合成黄体酮R5020共处理可逆转对lh诱导的体外培养排卵前卵泡中Pacap基因表达的抑制作用。我们进一步克隆了大鼠Pacap基因的5'-侧翼区域，并确定了共识孕酮受体元件的存在。将含有Pacap基因启动子的荧光素酶融合基因瞬时转染到排卵前卵泡颗粒细胞中，LH显著刺激了荧光素酶的活性。用RU486或依泊斯坦处理可部分抑制lh刺激的PACAP启动子活性。综上所述，黄体酮通过黄体酮受体作用于促性腺激素诱导排卵前卵泡颗粒细胞中Pacap基因的表达，从而可能参与排卵过程。
The present study was designed to determine whether progesterone might have a role in gonadotrophin-induced pituitary adenylate cyclase-activating polypeptide (Pacap) gene expression in rat ovary. Northern blot analysis revealed that treatment of pregnant mare's serum gonadotrophin (PMSG)-primed immature rats with the progestin antagonist RU486 or an inhibitor of 3beta-hydroxysteroid dehydrogenase epostane, 1 h before HCG, resulted in a dose-dependent inhibition of the HCG-induced Pacap gene expression. In-situ hybridization demonstrated that the number of pre-ovulatory follicles expressing Pacap mRNA in their granulosa cells was greatly reduced in ovaries treated with RU486. Moreover, the suppressive effect of RU486 or epostane on the LH-induced Pacap gene expression in cultured pre-ovulatory follicles was reversed by co-treatment with the synthetic progestin R5020. We further cloned the 5'-flanking region of the rat Pacap gene and identified the presence of a consensus progesterone receptor element. When luciferase fusion genes containing Pacap gene promoter were transiently transfected into granulosa cells of pre-ovulatory follicles, luciferase activity was markedly stimulated by LH. Treatment with RU486 or epostane resulted in partial suppression of LH-stimulated PACAP promoter activity. Taken together, these results indicate that progesterone, acting through progesterone receptors, plays a role in gonadotrophin induction of Pacap gene expression in granulosa cells of pre-ovulatory follicles, and thereby may be involved in the process of ovulation.
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