Granulosa cell proliferation is inhibited by PGE2 in the primate ovulatory follicle.
Granulosa cell proliferation is inhibited by PGE2 in the primate ovulatory follicle.
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DOI：
10.1080/19768354.2020.1764385
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发表时间：
2020-05-21
影响因子：
2.9
通讯作者：
Duffy DM
中科院分区：
文献类型：
--
作者：
Lundberg PS;Moskowitz GJ;Bellacose C;Demirel E;Trau HA;Duffy DM
前列腺素E2（PGE 2）是排卵的关键旁分泌介质。很少有特定的PGE 2调控的基因产物已被确定，所以我们假设，PGE 2可能会调节蛋白质网络的表达和/或活性，以促进排卵。为了测试这一概念，使用不可逆性途径分析（IPA）来预测灵长类动物排卵卵泡中PGE 2调节的功能。食蟹猴接受卵巢刺激。卵泡颗粒细胞在排卵剂量的人绒毛膜促性腺激素（hCG）之前（0小时）或之后36小时获得，预期排卵在hCG之后37-40小时。颗粒细胞获得额外的猴子治疗后36小时，hCG和PTGS 2抑制剂塞来昔布，显着减少hCG刺激的卵泡前列腺素合成。通过微阵列测定颗粒细胞RNA表达，并使用IPA进行分析。与仅使用hCG相比，未发现颗粒细胞mRNA被hCG +塞来昔布显着上调或下调。然而，IPA预测，前列腺素消耗显着调节几个功能途径。选择细胞周期/细胞增殖进行进一步研究，因为已知减少的颗粒细胞增殖对于排卵和形成功能齐全的黄体是必要的。前瞻性的体内和体外实验证实了预测，hCG刺激的颗粒细胞增殖的停止是通过PGE 2介导的。我们的研究表明，PGE 2通过不涉及关键细胞周期调节因子的mRNA水平的显著调节的机制提供了关键的颗粒细胞增殖调节。通路分析正确地预测，PGE 2作为一个旁分泌介导的卵巢结构和功能的这一重要转变。
Prostaglandin E2 (PGE2) is a key paracrine mediator of ovulation. Few specific PGE2-regulated gene products have been identified, so we hypothesized that PGE2 may regulate the expression and/or activity of a network of proteins to promote ovulation. To test this concept, Ingenuity Pathway Analysis (IPA) was used to predict PGE2-regulated functionalities in the primate ovulatory follicle. Cynomolgus macaques underwent ovarian stimulation. Follicular granulosa cells were obtained before (0 h) or 36 h after an ovulatory dose of human chorionic gonadotropin (hCG), with ovulation anticipated 37–40 h after hCG. Granulosa cells were obtained from additional monkeys 36 h after treatment with hCG and the PTGS2 inhibitor celecoxib, which significantly reduced hCG-stimulated follicular prostaglandin synthesis. Granulosa cell RNA expression was determined by microarray and analyzed using IPA. No granulosa cell mRNAs were identified as being significantly up-regulated or down-regulated by hCG + celecoxib compared with hCG only. However, IPA predicted that prostaglandin depletion significantly regulated several functional pathways. Cell cycle/cell proliferation was selected for further study because decreased granulosa cell proliferation is known to be necessary for ovulation and formation of a fully-functional corpus luteum. Prospective in vivo and in vitro experiments confirmed the prediction that hCG-stimulated cessation of granulosa cell proliferation is mediated via PGE2. Our studies indicate that PGE2 provides critical regulation of granulosa cell proliferation through mechanisms that do not involve significant regulation of mRNA levels of key cell cycle regulators. Pathway analysis correctly predicted that PGE2 serves as a paracrine mediator of this important transition in ovarian structure and function.