ICAR-Indian Veterinary Research Institute, Physiology & Climatology Division, Izatnagar, Bareilly, Uttar Pradesh, 243122, India.
ICAR-Indian Veterinary Research Institute, Division of Animal Genetics, Izatnagar, Bareilly, Uttar Pradesh, 243122, India.
Sci Rep. 2020 Apr 22;10(1):6849. doi: 10.1038/s41598-020-63804-z.
The EGR family comprises of EGR 1, EGR 2, EGR 3 and EGR 4 which are involved in the transactivation of several genes. A broad range of extracellular stimuli by growth factors is capable of activating EGR mediated transactivation of genes involved in angiogenesis and cell proliferation. However, their role in controlling VEGF A and FGF 2 signaling in the CL of water buffalo is not known. The present study was conducted to understand the role of EGR mediated regulation of VEGF A and FGF 2 signaling in buffalo luteal cells. Towards this goal, luteal cells were cultured and treated with VEGF A and FGF 2 and the mRNA expression pattern of EGR family members were documented. The EGR 1 message was found to be up-regulated in luteal cells of buffalo at 72 hours of culture. The functional validation of EGR 1 gene was accomplished by knocking out (KO) of EGR 1 in cultured luteal cells by CRISPR/Cas9 mediated gene editing technology. The EGR 1 KO cells were then cultured and stimulated with VEGF A and FGF 2. It was observed that VEGF A and FGF 2 induced angiogenesis, cell proliferation and steroidogenesis in wild type luteal cells, whereas the response of the growth factors was attenuated in the EGR 1 KO cells. Taken together our study provides evidence convincingly that both VEGF and FGF mediate their biological action through a common intermediate, EGR 1, to regulate corpus luteum function of buffalo.
EGR 家族由 EGR1、EGR2、EGR3 和 EGR4 组成，它们参与了几个基因的反式激活。生长因子的广泛的细胞外刺激能够激活 EGR 介导的参与血管生成和细胞增殖的基因的反式激活。然而，它们在控制水牛 CL 中 VEGF A 和 FGF2 信号的作用尚不清楚。本研究旨在探讨 EGR 介导的 VEGF A 和 FGF2 信号在水牛黄体细胞中的调控作用。为此，培养黄体细胞并用 VEGF A 和 FGF2 处理，记录 EGR 家族成员的 mRNA 表达模式。结果发现，在培养的 72 小时时，水牛黄体细胞中的 EGR1 信使上调。通过 CRISPR/Cas9 介导的基因编辑技术敲除（KO）培养的黄体细胞中的 EGR1 基因，完成了 EGR1 基因的功能验证。然后培养 EGR1 KO 细胞并用 VEGF A 和 FGF2 刺激。结果发现，VEGF A 和 FGF2 诱导野生型黄体细胞的血管生成、细胞增殖和类固醇生成，而在 EGR1 KO 细胞中，生长因子的反应减弱。总之，我们的研究提供了令人信服的证据，证明 VEGF 和 FGF 都通过共同的中间物 EGR1 来调节水牛黄体功能来发挥其生物学作用。
