调控牛卵巢颗粒细胞中转录因子 E2F1 mRNA。
Regulation of the transcription factor E2F1 mRNA in ovarian granulosa cells of cattle.
机构信息
Department of Animal and Food Sciences, Oklahoma State University, Stillwater, OK.
出版信息
J Anim Sci. 2020 Jan 1;98(1). doi: 10.1093/jas/skz376.
The E2F family of transcription factors plays an important role in the control of the cell cycle, cell proliferation, and differentiation, and their role in ovarian function is just emerging. Although some evidence suggests a possible role of E2F1 in ovarian follicular development, what regulates its production in ovarian cells is unknown. Objectives of this study were to determine whether: (i) E2F1 gene expression in granulosa cells (GCs) and theca cells (TCs) change with follicular development and (ii) E2F1 mRNA abundance in TC and GC is hormonally regulated. Using real-time PCR, E2F1 mRNA abundance in GC was 5.5-fold greater (P < 0.05) in small (SM; 1 to 5 mm) than large (LG; >8 mm) follicles, but in TC, E2F1 expression did not differ among follicle sizes. SM-follicle GC had 2.1-fold greater (P < 0.05) E2F1 mRNA than TC. In SM-follicle GC, FGF9 induced a 7.6-fold increase in E2F1 mRNA abundance; however, FGF9 did not affect (P > 0.10) abundance of E2F1 mRNA in LG-follicle TC or GC. Follicle-stimulating hormone (FSH) had no effect (P > 0.10) on E2F1 gene expression in SM- or LG-follicle GC. SM-follicle GC were concomitantly treated with insulin-like growth factor 1 (30 ng/mL), FSH (30 ng/mL), and either 0 or 30 ng/mL of FGF9 with or without 50 µM of an E2F inhibitor (E2Fi; HLM0064741); FGF9 alone increased (P < 0.05) GC numbers, whereas E2Fi alone decreased (P < 0.05) GC numbers, and concomitant treatment of E2Fi with FGF9 blocked (P < 0.05) this stimulatory effect of FGF9. Estradiol production was inhibited (P < 0.05) by FGF9 alone and concomitant treatment of E2Fi with FGF9 attenuated (P < 0.05) this inhibitory effect of FGF9. SM-follicle GC treated with E2Fi decreased (P < 0.05) E2F1 mRNA abundance by 70%. Collectively, our studies show that GC E2F1 mRNA is developmentally and hormonally regulated in cattle. Inhibition of E2F1 reduced FGF9-induced GC proliferation and attenuated FGF9-inhibited estradiol production, indicating that E2F1 may be involved in follicular development in cattle.
E2F 转录因子家族在细胞周期、细胞增殖和分化的控制中发挥着重要作用，其在卵巢功能中的作用才刚刚显现。虽然有证据表明 E2F1 可能在卵巢卵泡发育中发挥作用，但调节其在卵巢细胞中产生的因素尚不清楚。本研究的目的是确定：（i）颗粒细胞（GC）和膜细胞（TC）中 E2F1 基因表达随卵泡发育而变化；（ii）TC 和 GC 中的 E2F1 mRNA 丰度是否受激素调节。通过实时 PCR，小（SM；1 至 5 毫米）卵泡中的 GC 中 E2F1 mRNA 丰度比大（LG；>8 毫米）卵泡高 5.5 倍（P<0.05），但 TC 中卵泡大小之间的 E2F1 表达没有差异。SM 卵泡 GC 的 E2F1 mRNA 丰度比 TC 高 2.1 倍（P<0.05）。在 SM 卵泡 GC 中，FGF9 诱导 E2F1 mRNA 丰度增加 7.6 倍；然而，FGF9 对 LG 卵泡 TC 或 GC 中 E2F1 mRNA 的丰度没有影响（P>0.10）。卵泡刺激素（FSH）对 SM 或 LG 卵泡 GC 中的 E2F1 基因表达没有影响（P>0.10）。SM 卵泡 GC 同时用胰岛素样生长因子 1（30ng/mL）、FSH（30ng/mL）和 0 或 30ng/mL 的 FGF9 以及 E2F 抑制剂（E2Fi；HLM0064741）进行处理，或不处理；单独的 FGF9 增加了（P<0.05）GC 数量，而单独的 E2Fi 减少了（P<0.05）GC 数量，并且 E2Fi 与 FGF9 的同时处理阻断了（P<0.05）FGF9 的这种刺激作用。FGF9 单独抑制（P<0.05）雌二醇的产生，E2Fi 与 FGF9 的同时处理减弱了（P<0.05）FGF9 的这种抑制作用。用 E2Fi 处理的 SM 卵泡 GC 中 E2F1 mRNA 丰度降低了 70%（P<0.05）。总的来说，我们的研究表明，牛的 GC E2F1 mRNA 是发育和激素调节的。E2F1 的抑制减少了 FGF9 诱导的 GC 增殖，并减弱了 FGF9 抑制的雌二醇产生，表明 E2F1 可能参与了牛的卵泡发育。
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