Epigenetic modification cooperates with Zeb1 transcription factor to regulate Bmp4 to promote chicken PGCs formation
Epigenetic modification cooperates with Zeb1 transcription factor to regulate Bmp4 to promote chicken PGCs formation
复制标题
表观遗传修饰配合Zeb1转录因子调控Bmp4促进鸡PGCs形成
DOI：
10.1016/j.gene.2021.145760
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发表时间：
2021-06-17
期刊：
影响因子：
3.5
通讯作者：
Li, Bichun
中科院分区：
文献类型：
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作者：
Zhou, Shujian;Li, Tingting;Li, Bichun
BMP4是哺乳动物原始生殖细胞形成的关键基因，但鸡PGCs形成的机制尚不清楚。在这项研究中，我们比较了不同物种的进化关系。尽管蛋白质序列在小鼠、人类和鸡之间高度保守，但启动子在鸟类和哺乳动物物种之间存在差异。因此，很容易预测鸡体内Bmp4表达可能存在不同的调控机制。在这里，我们阐明了鸡 Bmp4 在 PGC 形成过程中的功能。在体内，Bmp4可以促进PGCs的发育和迁移，并增加关键基因（Cvh、c-kit、cxcr4等）的表达。而敲除 Bmp4 后，这些基因的表达将会降低。体外过表达和敲除Bmp4后发现，过表达Bmp4可以促进拟胚体（EB）的形成并上调PGC形成和迁移的关键基因，而敲除Bmp4可以抑制拟胚体的形成并降低相关基因的表达。流式细胞术和间接免疫荧光法也显示出相同的结果。这些结果证明鸡Bmp4也能促进PGCs的形成。此外，双荧光素酶活性检测表明Bmp4的启动子活性受到转录因子Zeb1的正向调控。 Zeb1 的过表达还可以增加 Bmp4 的 mRNA 和蛋白表达。同时，DNA甲基化抑制Bmp4转录，组蛋白甲基化能够促进其转录。总之，本研究证实鸡Bmp4可以促进鸡PGC的形成。该基因受 DNA、组蛋白甲基化和转录因子 Zeb1 调节。这些结果为探讨Bmp4在PGCs形成过程中的功能和分子机制奠定了理论基础。
BMP4 is the critical gene of primordial germ cell formation in mammal, however, the mechanism of PGCs formation in chicken still unknown. In this research, we compared the evolution relationship of different species. Although the protein sequence is highly conservative between mouse, human and chicken, promotors vary among avian and mammal species. Therefore, it is easily to predict that there would be different regulation mechanism of Bmp4 expression in chicken. Here, we elucidate the function of chicken Bmp4 during PGCs formation. In vivo, Bmp4 can promote PGCs development and migration, and increase the expression of key genes (Cvh, c-kit, cxcr4, etc.). Whereas, the expression of these genes will decrease after knocking out Bmp4. After overexpression and knockout Bmp4 in vitro, we found that overexpression of Bmp4 could promote the formation of embryoid bodies (EB) and up-regulate the key genes of PGCs formation and migration, while knockout Bmp4 could inhibit the formation of embryoid bodies and decrease the expression of related genes. Flow and indirect immunofluorescence also indicated the same result. These all results proved that chicken Bmp4 could also promote the formation of PGCs. Furthermore, dual-luciferase activity detection showed that the promotor activity of Bmp4 was positively regulated by transcription factor Zeb1. Overexpression of Zeb1 can also increase the mRNA and protein expression of Bmp4. At the same time, DNA methylation inhibited Bmp4 transcription and histone methylation was able to promote its transcription. In conclusion, this study established that chicken Bmp4 can promote the formation of chicken PGCs. This gene is regulated by DNA, histone methylation and transcription factor Zeb1. These results lay a theoretical foundation for exploring the function and molecular mechanism of Bmp4 in the process of PGCs formation.