GATA4 and GATA6 Silencing in Ovarian Granulosa Cells Affects Levels of mRNAs Involved in Steroidogenesis, Extracellular Structure Organization, IGF-I Activity, and Apoptosis
GATA4 and GATA6 Silencing in Ovarian Granulosa Cells Affects Levels of mRNAs Involved in Steroidogenesis, Extracellular Structure Organization, IGF-I Activity, and Apoptosis
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DOI：
10.1210/en.2013-1410
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发表时间：
2013-12-01
期刊：
影响因子：
4.8
通讯作者：
Stocco, Carlos
中科院分区：
文献类型：
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作者：
Bennett, Jill;Baumgarten, Sarah C.;Stocco, Carlos
颗粒细胞 (GC) 中转录因子 GATA4 和 GATA6 的敲低会损害卵泡发生并诱导不孕。为了研究这些因素调控的途径和基因，我们对野生型 GC 或缺乏 GATA4、GATA6 或 GATA4/6（G4（gcko）、G6（gcko）和 G4/6（gcko））的 GC 在体内用马绒毛膜促性腺激素处理后进行了微阵列分析。 GATA4 缺失影响的基因数量比 GATA6 多，这与在 G4(gcko) 小鼠中观察到的生育力低下以及在 G6(gcko) 动物中发现的正常生殖功能相关。删除这两个因子会影响更多的基因。此外，与单一 GATA 缺陷动物相比，已知对卵巢功能至关重要的 FSH 受体、LH 受体、抑制素 α 和 β、多功能蛋白聚糖、妊娠相关血浆蛋白 A 以及蛋白激酶 A 调节单元 2b 的表达在 GATA4 和 GATA6 双重敲除中受到极大影响。这表明 GATA4 和 GATA6 在关键卵巢基因的调节中功能上相互补偿。功能丰富表明，G4/6（gcko）小鼠的排卵、生长、细胞内信号传导、细胞外结构组织、促性腺激素和生长因子作用以及类固醇生成均受到显着调节。该分析的结果通过定量聚合酶链反应、免疫组织化学和生物测定得到证实。用 cAMP/IGF-I 处理 GC，以绕过 FSH 和 IGF-I 信号传导缺陷，结果表明大多数受影响的基因是 GATA4/6 的直接靶标。受 GATA 敲低影响的途径的多样性强调了这些因素在 GC 功能调节中的重要作用。
Knockdown of the transcription factors GATA4 and GATA6 in granulosa cells (GCs) impairs folliculogenesis and induces infertility. To investigate the pathways and genes regulated by these factors, we performed microarray analyses on wild-type GCs or GCs lacking GATA4, GATA6, or GATA4/6 (G4(gcko), G6(gcko), and G4/6(gcko)) after in vivo treatment with equine chorionic gonadotropin. GATA4 deletion affected a greater number of genes than GATA6, which correlates with the subfertility observed in G4(gcko) mice and the normal reproductive function found in G6(gcko) animals. An even greater number of genes were affected by the deletion of both factors. Moreover, the expression of FSH receptor, LH receptor, inhibin alpha and beta, versican, pregnancy-associated plasma protein A, and the regulatory unit 2b of protein kinase A, which are known to be crucial for ovarian function, was greatly affected in double GATA4 and GATA6 knockouts when compared with single GATA-deficient animals. This suggests that GATA4 and GATA6 functionally compensate for each other in the regulation of key ovarian genes. Functional enrichment revealed that ovulation, growth, intracellular signaling, extracellular structure organization, gonadotropin and growth factor actions, and steroidogenesis were significantly regulated in G4/6(gcko) mice. The results of this analysis were confirmed using quantitative polymerase chain reaction, immunohistochemical, and biological assays. Treatment of GCs with cAMP/IGF-I, to bypass FSH and IGF-I signaling defects, revealed that most of the affected genes are direct targets of GATA4/6. The diversity of pathways affected by the knockdown of GATA underscores the important role of these factors in the regulation of GC function.