| 徐瑜珊,陈 丽,王 娜,臧春燕,彭凡珂,张 军.抑制糖酵解活性限制雄性原始生殖细胞的增殖[J].,2019,19(10):1823-1828 |
| 抑制糖酵解活性限制雄性原始生殖细胞的增殖 |
| Inhibition of Glycolysis Activity Limits the Proliferation of Male Primordial Germ Cells |
| 投稿时间：2019-01-14 修订日期：2019-02-05 |
| DOI：10.13241/j.cnki.pmb.2019.10.005 |
| 中文关键词: 雄性生殖细胞 糖代谢 多能性 分化 |
| 英文关键词: Male primordial germ cell Glycolysis Pluripotency Differentiation |
| 基金项目:国家自然科学基金项目（31871445）；国家自然科学基金项目（31501211） |
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| 中文摘要: |
| 摘要 目的：雄性原始生殖细胞在植入生殖嵴后，会从有丝分裂退出进入静息状态，在这一过程中伴随着细胞内代谢状态的改变，本研究旨在体解析原始生殖细胞增殖的改变与细胞代谢之间的因果关系。方法：通过体内Brdu掺入实验明确不同时间点雄性生殖细胞的增殖状态；分析比较增殖状态和静息状态原始生殖细胞糖酵解相关基因的表达；利用腹腔注射HK2特异性抑制剂2-Deoxy-D-glucose (2-DG)，构建糖酵解抑制小鼠模型；通过免疫荧光与qPCR分析抑制糖酵解后原始生殖细胞的表型。结果：免疫荧光结果显示雄性生殖细胞增殖停滞从E13.5开始，至E15.5完全停滞； qPCR和Western Blot显示在此过程中HK2的表达是逐渐降低的；在E11.5抑制小鼠胚胎中的糖酵解过程，可以在E13.5检测到雄性PGCs增殖下降，并且可以抑制多能性基因如Sox2、Oct4的表达。结论：研究发现，E11.5-E13.5 雄性原始生殖细胞内增殖与多能性的维持需要糖酵解。改变胚胎糖酵解水平可以影响原始生殖细胞增殖分化进程。 |
| 英文摘要: |
| ABSTRACT Objective: Male primordial germ cells will exit from mitosis and enter a cell cycle arrest after implantation of genital ridge, accompanied by changes in intracellular metabolic states. Our work is to explore causal relationship between primordial germ cells proliferation and cell metabolism. Methods: The proliferation states of male primordial germ cells at different time points was determined by BrdU incorporation assay. The expression of glycolysis related genes of primordial germ cells in the proliferative state and cell cycle arrest state was compared by qPCR. A mouse model of glycolysis inhibition was established by intraperitoneal injection of HK2-specific inhibitor 2-DG, and phenotype was analyzed by immunofluorescence and qPCR. Results: The results of immunofluorescence showed that the cell cycle arrest of male germ cells start at E13.5 and completely arrest at E15.5. qPCR and Western Blot showed that Hk2 was decreased during this process. Inhibition of Hk2 in mouse embryos at E11.5, the proliferation of male germ cells was significantly inhibited at E13.5, and the expression levels of pluripotent genes such as Sox2 and Oct4 were decreased. Conclusion: This study found that in E11.5-E13.5, glycolysis is required for proliferation and maintenance of pluripotency in male primordial germ cells. Changing the level of embryonic glycolysis can affect the proliferation and differentiation of primordial germ cells. |
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