Growth differentiation factor-9 stimulates progesterone synthesis in granulosa cells via a prostaglandin E2/EP2 receptor pathway
Growth differentiation factor-9 stimulates progesterone synthesis in granulosa cells via a prostaglandin E2/EP2 receptor pathway
复制标题
DOI：
10.1073/pnas.180295197
复制
发表时间：
2000-08-29
影响因子：
11.1
通讯作者：
Matzuk, MM
中科院分区：
文献类型：
--
作者：
Elvin, JA;Yan, CN;Matzuk, MM
生长分化因子 9 (GDF-9)、卵母细胞分泌的转化生长因子 β 超家族成员、黄体酮受体、环氧合酶 2 (Cox2; Ptgs2) 和 EP2 前列腺素 E-2 (PGE(2)) 受体 (EP2; Ptgerep2) 是雌性而非雄性小鼠生育能力所必需的。为了确定这些因素的相互关系，我们使用了排卵前颗粒细胞培养系统，其中添加了重组 GDF-9、前列腺素、前列腺素受体激动剂或环氧合酶抑制剂。 GDF-9在2小时内刺激Cox2 mRNA，在6小时内刺激PGE(2)；然而，直到添加GDF-9后12小时黄体酮才增加。这表明 Cox2 是 CDF-9 的直接下游靶标，但孕酮合成需要中间体。为了确定黄体酮的产生是否需要前列腺素合成，我们分析了 PGE(2) 和环氧合酶抑制剂对该过程的影响。 PGE(2) 本身可以刺激黄体酮合成，但不如 GDF-9 有效（24 小时内分别增加 3 倍和 6 倍）。此外，吲哚美辛或 NS-398（Cox2 抑制剂）可阻断基础和 GDF-9 刺激的孕酮合成。然而，在含有 GDF-9 和 NS-398 的培养物中添加 PGE(2) 会覆盖黄体酮合成中 NS-398 的阻滞。为了进一步定义 PGE(2) 依赖性途径，我们证明布他前列素（一种特定的 EP2 激动剂）可刺激孕酮合成并覆盖 NS-398 阻滞。此外，CDF-9 通过前列腺素和黄体酮独立途径刺激 EP2 mRNA 合成。因此，CDF-9诱导EP2信号转导途径，这似乎是颗粒卵丘细胞中孕酮合成所必需的。这些研究进一步证明了卵母细胞-体细胞相互作用在女性生殖中的重要性。
Growth differentiation factor-9 (GDF-9), an oocyte-secreted member of the transforming growth factor beta superfamily, progesterone receptor, cyclooxygenase 2 (Cox2; Ptgs2), and the EP2 prostaglandin E-2 (PGE(2)) receptor (EP2; Ptgerep2) are required for fertility in female but not male mice. To define the interrelationship of these factors, we used a preovulatory granulosa cell culture system in which we added recombinant GDF-9, prostaglandins, prostaglandin receptor agonists, or cyclooxygenase inhibitors. GDF-9 stimulated Cox2 mRNA within 2 h, and PGE(2) within 6 h; however, progesterone was not increased until 12 h after addition of GDF-9. This suggested that Cox2 is a direct downstream target of CDF-9 but that progesterone synthesis required an intermediate. To determine whether prostaglandin synthesis was required for progesterone production, we analyzed the effects of PGE(2) and cyclooxygenase inhibitors on this process. PGE(2) can stimulate progesterone synthesis by itself, although less effectively than GDF-9 (3-fold vs. 6-fold increase over 24 h, respectively). Furthermore, indomethacin or NS-398, inhibitors of Cox2, block basal and GDF-9-stimulated progesterone synthesis. However, addition of PGE(2) to cultures containing both GDF-9 and NS-398 overrides the NS-398 block in progesterone synthesis. To further define the PGE(2)-dependent pathway, we show that butaprost, a specific EP2 agonist, stimulates progesterone synthesis and overrides the NS-398 block. In addition, CDF-9 stimulates EP2 mRNA synthesis by a prostaglandin- and progesterone-independent pathway. Thus, CDF-9 induces an EP2 signal transduction pathway which appears to be required for progesterone synthesis in cumulus granulosa cells. These studies further demonstrate the importance of oocyte-somatic cell interactions in female reproduction.