Drp1对PINK1缺失果蝇保护机制的研究

发布时间：2018-04-23 11:37

本文选题：帕金森病 + 果蝇　；参考：《中南大学》2014年硕士论文

【摘要】：帕金森病(Parkinson's Disease, PD)是一种最常见的神经退行性运动障碍疾病。线粒体功能损伤是PD的重要致病机制之一。目前己鉴定了16个与PD相关的基因,其中PINK1突变会导致常染色体隐性遗传的PD。PINK1是一个可以锚定在线粒体上的丝氨酸/苏氨酸激酶。大量的研究表明PINK1与线粒体的功能密切相关。PINK1缺失的果蝇表现为运动障碍及线粒体结构和功能异常。线粒体需要通过不断的相互融合和分裂维持其正常功能,而这一过程的紊乱会导致线粒体结构和功能异常,进一步引起细胞损伤。Drp1是线粒体分裂相关蛋白。在果蝇里进行的遗传互作研究表明,过表达Drp1增加线粒体的分裂可以恢复PINK1缺失导致的异常表型。然而Drp1恢复PINK1缺失导致的相关表型的具体机制还不是很清楚。我们的前期研究发现,PINK1敲除的小鼠中Drpl596位点丝氨酸的磷酸化会发生改变。因此,本论文将进一步探讨Drp1S596位点的磷酸化对PINK1缺失果蝇的影响。目的：探讨Drp1S596位点的磷酸化对PINK1缺失果蝇的影响。方法：在PINK1缺失果蝇背景下分别过表达Drp1WT, Drp1S596A(模拟去磷酸化)和Drp1S596D(模拟磷酸化),观察Drp1S596位点的磷酸化对PINK1缺失导致的异常表型的影响。结果：Drp1S596D和Drp1WT均可以恢复PINK1缺失果蝇的异常表型,包括翅膀姿势异常、背部塌陷、运动能力下降、肌肉和多巴胺能神经元线粒体形态异常以及ATP生成减少；而Drp1S596A不能使PINK1缺失果蝇的运动能力及肌肉和多巴胺能神经元中的线粒体形态恢复正常,但是Drp1S596A也能使翅膀异常、ATP生成水平恢复正常。结论：Drp1可能通过S596位点的磷酸化来参与调节PINK1介导的线粒体的形态和功能。这为阐明Drp1恢复PINK1缺失果蝇异常表型的具体机制,进一步了解PD的发病机制提供了实验依据。
[Abstract]:Parkinson's disease (PD) is one of the most common neurodegenerative motor disorders. Mitochondrial dysfunction is one of the important pathogenetic mechanisms of PD. Sixteen PD-related genes have been identified, of which PINK1 mutation leads to autosomal recessive inheritance of PD.PINK1, a serine / threonine kinase anchored on mitochondria. A large number of studies have shown that Drosophila melanogaster, which is closely related to the function of mitochondria and the deletion of PINK1, shows dyskinesia and abnormal structure and function of mitochondria. Mitochondria need to maintain their normal function through continuous fusion and division, and the disorder of this process will lead to abnormal structure and function of mitochondria, and further cause cell damage. Drp1 is a mitosynthesis-related protein of mitochondria. Genetic interaction studies in Drosophila have shown that overexpression of Drp1 increases mitochondrial division and restores abnormal phenotypes caused by PINK1 deletion. However, the specific mechanism of Drp1 recovery of the associated phenotypes resulting from PINK1 deletion is not well understood. Our previous study found that Drpl596 site serine phosphorylation changes in PINK1 knockout mice. Therefore, the effect of phosphorylation of Drp1S596 site on PINK1 deletion in Drosophila melanogaster is further investigated. Objective: to investigate the effect of phosphorylation of Drp1S596 site on PINK1 deletion in Drosophila melanogaster. Methods: Drp1WT, Drp1S596A (simulated dephosphorylation) and Drp1S596D (simulated phosphorylation) were overexpressed under the background of PINK1 deletion in Drosophila melanogaster respectively. The effects of phosphorylation of Drp1S596 site on abnormal phenotypes induced by PINK1 deletion were observed. Results both 1 / Drp1S596D and Drp1WT could restore the abnormal phenotypes of Drosophila PINK1 deletion, including abnormal wing posture, back collapse, decreased motor ability, abnormal mitochondrial morphology of muscle and dopaminergic neurons, and decreased ATP production. However, Drp1S596A could not restore the motor ability of Drosophila melanogaster PINK1 deficiency and mitochondrial morphology in muscle and dopaminergic neurons, but Drp1S596A could also restore the level of ATP production in abnormal wings of Drosophila melanogaster. Conclusion it is suggested that the phosphorylation of S596 site may be involved in regulating the morphology and function of mitochondria mediated by PINK1. This provides experimental evidence for elucidating the specific mechanism of Drp1 restoring the abnormal phenotype of PINK1 deletion Drosophila and further understanding the pathogenesis of PD.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R742.5

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