Growth differentiation factor 9 promotes follicle-stimulating hormone-induced progesterone production in chicken follicular granulosa cells
Growth differentiation factor 9 promotes follicle-stimulating hormone-induced progesterone production in chicken follicular granulosa cells
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生长分化因子 9 促进鸡卵泡颗粒细胞中卵泡刺激素诱导的黄体酮产生
DOI：
10.1016/j.ygcen.2019.03.005
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发表时间：
2019-05-15
影响因子：
2.7
通讯作者：
Gong, Yanzhang
中科院分区：
文献类型：
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作者：
Li, Jinqiu;Luo, Wei;Gong, Yanzhang
与哺乳动物中的已知功能相比，迄今为止，鸟类中卵母细胞源性生长分化因子 9 (GDF9) 在卵泡中的功能尚不清楚。我们的目的是研究 GDF9 对类固醇生成和鸡卵巢颗粒细胞 (GC) 有丝分裂的影响。来自分级前（直径 6-8 mm，phGC）和排卵前卵泡（F1-F5，poGC）的原代 GC 在存在或不存在 GDF9 蛋白的情况下进行培养。然后通过放射免疫分析(RIA)测定培养基中的孕酮(P4)水平，并通过定量PCR检测类固醇生成基因的表达水平。我们发现单独的 GDF9 通过调节 STAR、CYP11A1 和 HSD3B 等类固醇生成基因的表达对 P4 水平没有显着影响。进一步的实验表明，GDF9 促进卵泡刺激素 (FSH) 诱导的 P4 产生和 STAR 表达。 GDF9还挽救了FSH诱导的FSH受体（FSHR）表达减少，但对毛喉素诱导的P4、STAR和毛喉素抑制的FSHR表达水平没有影响，表明GDF9可能通过增强FSHR和STAR表达来实现其对P4的调节作用。此外，GDF9还促进GC细胞周期进程，调节相关基因的基因转录，增强DNA复制并抑制细胞凋亡。有趣的是，这些效应在 phGC 和 poGC 之间是不同的。据我们所知，这是第一份说明 GDF9 对鸡 GC 的功能以及对卵巢类固醇生成的影响的报告。我们的研究结果强调了中央卵母细胞对周围颗粒细胞的调节，并强调了旁分泌信号和内分泌激素对卵巢黄体酮产生的相互作用；这些发现有助于了解禽类卵巢卵泡的发育。
The function of oocyte-derived growth differentiation factor 9 (GDF9) in ovarian follicles has thus far been poorly defined in avian species compared with the defined function in mammals. Our aim here is to investigate the effects of GDF9 on steroidogenesis and on chicken ovarian granulosa cell (GC) mitosis. Primary GCs from both prehierarchical (6-8 mm in diameter, phGCs) and preovulatory follicles (F1-F5, poGCs) were cultured in the presence or absence of the GDF9 protein. The progesterone (P4) levels in the culture medium were then measured by radioimmunoassay (RIA), and the expression levels of steroidogenesis genes were detected by quantitative PCR. We found that GDF9 alone showed no significant effect on the P4 levels by regulating the expression of steroidogenesis genes, such as STAR, CYP11A1 and HSD3B. Further experiments indicated that GDF9 promoted follicle-stimulating hormone (FSH)-induced P4 production and STAR expression. GDF9 also rescued the FSH-induced decrease of FSH receptor (FSHR) expression but had no effect on the forskolin-induced P4, STAR and forskolin-inhibited FSHR expression levels, suggesting that GDF9 might achieve its regulatory role of P4 by enhancing FSHR and STAR expression. In addition, GDF9 also promoted GC cell cycle progression, regulated the gene transcription of related genes, potentiated DNA replication and inhibited apoptosis. Interestingly, these effects differed between the phGCs and the poGCs. To our knowledge, this is the first report that illustrates the function of GDF9 on chicken GCs and the effects on ovarian steroidogenesis. Our findings highlight the regulation of central oocytes on the surrounding granulosa cells and emphasize the interaction between paracrine signals and endocrine hormones on ovarian progesterone production; these findings contribute to the understanding of the development of avian ovarian follicles.