ETS1对非洲爪蛙早期胚胎发育的影响
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国家自然科学基金(81200125);江苏高校优势学科建设工程资助项目;江苏省“青蓝工程”(2010)


Effects of ETS1 on early embryo development of Xenopus laevis
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    摘要:

    目的:通过在非洲爪蛙胚胎中过表达ETS1探讨该基因对爪蛙胚胎早期发育的影响-方法:将克隆PCR获得的ETS1全长片段连接到质粒上,获得pCS2+-ETS1质粒;利用RT-PCR和Western blot检测ETS1 mRNA和蛋白在胚胎发育不同时期的表达情况;显微注射ETS1 mRNA入爪蛙胚胎,利用整胚原位杂交-定量PCR检测胚层标志性基因的表达;通过Western blot检测过表达ETS1对细胞凋亡和增殖的影响-结果:ETS1从受精卵时期即已开始表达,在器官形成期表达增强,过表达ETS1抑制了细胞分化以及神经胚期中胚层和外胚层标志基因表达,凋亡相关蛋白表达增加,增殖相关蛋白无明显变化-结论:ETS1表达于非洲爪蛙胚胎发育各阶段,过表达ETS1抑制了中胚层和外胚层发育,促进细胞凋亡但不影响细胞增殖-

    Abstract:

    Objective:To investigate effects of ETS1 on early embryo development of Xenopus laevis. Methods:The pCS2+-ETS1 plasmid was produced by ligating pCS2+ plasmid and ETS1 cloned according to PCR. The expressions of the ETS1 during embryogenesis were detected by RT-PCR and Western blot. After micro-injection of ETS1 mRNA into embryo,the expressions of marker genes of different embryonic layers were detected by whole mount in situ hybridization and quantitative RT-PCR. The effect of ETS1 over-expression on apoptosis and proliferation of cells was analyzed by Western blot. Results:The expression of ETS1 started from the period of oosperm,and increased through all periods of organogenesis. The overexpression of ETS1 inhibited differentiation of cells and expression of marker genes of mesoderm and ectoderm in neurulation. The expression of apoptosis-related proteins was increased,whereas the expression of proliferation-related proteins had no significant change. Conclusion:ETS1 is expressed through all periods of embryogenesis of Xenopus laevis. The overexpression of ETS1 inhibits the development of mesoderm and ectoderm,as well as promotes the process of apoptosis,nonetheless no change of proliferation is inspected.

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殷晨阳,袁 芳,刘 佳,郭 静,袁 栎. ETS1对非洲爪蛙早期胚胎发育的影响[J].南京医科大学学报(自然科学版),2014,(3):291-296

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  • 收稿日期:2013-10-18
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  • 在线发布日期: 2014-03-26
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