the mechanism of cell metabolism regulation in necrosis

细胞程序性死亡是由细胞内信号决定的自主有序的细胞死亡方式，对正常细 胞群的稳定、机体的免疫反应、肿瘤的抑制等都有非常重要的作用。根据细胞死 亡形态学上的差异，可以将细胞死亡方式分为细胞凋亡和细胞坏死。 我们通过研究发现，细胞坏死过程中会发生细胞有氧呼吸的强烈激活，而细 胞凋亡过程不会发生。对细胞有氧呼吸的抑制可以明显抑制细胞坏死，而对细胞 凋亡没有作用，这表明细胞坏死和凋亡对细胞代谢的依赖程度具有明显差异。 RIP3作为细胞坏死过程中的关键调控因子，在细胞代谢的激活中具有重要作用。 在RIP1/RIP3/MLKL坏死复合体形成过程中，RIP3会通过直接磷酸化的方式激 活线粒体代...Programmed cell death is a kind of organizedcell suicide regulated by intracellular signals, which is very important to cell population homeostasis, immune response and tumorigenesis inhibition. According to the different morphology, programmed cell death can be divided into apoptosis and necroptosis. In our study we found that aerobic respiration was activated in necrotic cells, but not ...学位：理学博士院系专业：生命科学学院_生物化学与分子生物学学号：2162012015377

I-SceI Meganuclease Mediates Transgenesis in Zebrafish and Connexin48.5 Localization Research

通信作者:[email protected]细胞连接蛋白是由多基因家族编码的一类结构相似、分子质量不同的蛋白质.在细胞膜上,每6个相同或不同的连接蛋白围绕中央孔排列形成一个连接子,相邻细胞膜上的连接子相互对接形成细胞间隙连接,细胞间隙连接是一种重要的通讯连接,它不仅是细胞间代谢偶联、冲动传导的结构基础,而且可以通过介导与细胞的迁移、分化、增生和器官形成有关的信号物质而在胚胎发育中起重要作用.为了进一步探讨细胞间隙连接在生物体中的作用,采用斑马鱼胚胎显微注射核酸酶I-SceI和质粒DNA的方法成功构建了绿色荧光蛋白GFP与细胞连接蛋白Connexin48.5融合表达的转基因荧光斑马鱼品系,并通过对荧光蛋白发光的观察在斑马鱼鱼体中进行连接蛋白Connexin48.5的定位.实验结果表明:核酸酶I-SceI介导下的斑马鱼转基因方法效率较高,可行性好;连接蛋白Connexin48.5在斑马鱼体内主要定位于眼球晶状体和脊索.所获得的Connexin48.5-GFP融合表达斑马鱼系,以及利用该品系进行的Connexin48.5定位研究对细胞连接蛋白在生物体内的功能研究将具有重要作用.[英文文摘]Connexins are a group of proteins coded by multi gene family.On the cell membrane,every 6 Connexins combine together to form a Connexon,and two Connexons of two different cells connect with each other to form a gap junction.Gap junction is a very important kind of communicctional connection between cells,which is not only the constructional basic of metabolic coupling and signal transduction betweens cells,but also the channeel of signal substance controlling cell migration,differentiation,proliferation and organogenesis.For a deeper study of gap junction function in organisms,we generated a transgenic zebrafish line that expresses Connexin 48.5-GFP fusion protein by co-injecting meganuclease I-Scel and plasmid DNA into zebrafish embryos.I-Scel mediate transgenesis showed high efficiency,which made it a better way to get transgenic animals than the traditional method.While the working mechanism of I-Scel remains unclear,in our opinion,I-Scel binds to its recognition site after the cleavage and promotes unclear localization of the transgene could be a good explanation.Using our transgenic zebrafish line,we found that Connexin 48.5 is highly expressed in lens and notochord in zebrafish.In addition to its expression in lens,which has been reported preciously,our new finding suggested that Connexin48.5 could have important roles 　in notochord development or nervous system function.国家重点基础研究发展计划(973)项目(2009CB22200