Drp1、Marf对线粒体形态结构及其功能影响

发布时间：2018-02-27 00:28

本文关键词： 线粒体 NDUFS3 Drp1 Marf　出处：《桂林医学院》2017年硕士论文　论文类型：学位论文

【摘要】：目的:通过构建Drp1(dynamin-related protein 1)、Marf(mitoechondrial assembly regulatory factor)转基因果蝇,探讨Drp1、Marf对果蝇间接飞行肌的影响,以及Drp1、Marf对线粒体形态结构的调控及其功能影响的相关性研究。方法:使用MHC-Gal4启动子,利用经典的GAL4/UAS系统,使果蝇线粒体形态调控基因Drp1和Marf特异性表达于肌肉组织中。通过敲除Drp1和过表达Marf基因干预调控其在果蝇肌肉线粒体的表达,改变线粒体的形态,观察果蝇翅膀的异常形态以及飞行能力,并通过透射电子显微镜观察果蝇肌肉组织间接飞行肌肌肉的排列和线粒体的形态学是否改变。通过ATP试剂盒检测线粒体ATP产能功能,并进一步应用Western Blot蛋白免疫印迹技术检测线粒体氧化呼吸链复合物I亚单位NDUFS3蛋白量表达量,来探讨Drp1、Marf对线粒体线粒体功能的影响。结果:相比于对照组,通过敲除Drp1和过表达Marf基因均使果蝇间接飞行肌发生障碍,表现为飞行能力变差,翅膀的异常率增高;电镜可观察到Drp1、Marf的转基因果蝇胸部肌肉组织线粒体呈融合状态,形态结构受损,果蝇间接飞行肌肉排列不规则,肌纤维存在断裂。线粒体氧化呼吸产生的ATP含量减少以及Western Blot蛋白检测线粒体氧化呼吸复合物I亚单位NDUFS3蛋白表达含量均明显下调。结论:敲除Drp1和过表达Marf基因果蝇模型都可以改变线粒体的形态结构,影响果蝇的飞行功能;敲除Drp1和过表达Marf基因的果蝇模型都可以使线粒体功能下调;推测无论是线粒体分裂受限还是融合过度,都会使线粒体功能发生障碍。
[Abstract]:Objective: to construct Drp1 (dynamin-related protein 1), Marf (mitoechondrial assembly regulatory factor) to investigate the effects of transgenic Drosophila, Drp1, and Marf on Drosophila indirect flight muscle and Drp1, study the related regulation and functional effects of Marf on mitochondrial morphology. Methods: using the MHC-Gal4 promoter using the classical GAL4/UAS system the Drosophila mitochondrial morphology, gene expression and regulation of Drp1 and Marf in muscle tissue specificity. The deletion of Drp1 and overexpression of Marf gene intervention to regulate the expression in Drosophila muscle mitochondria, mitochondrial morphology change, observe the abnormal shape of the Drosophila wings and flying ability, and observed by transmission electron microscopy morphology of Drosophila muscle tissue indirectly the arrangement of flight muscle and mitochondria are changed. By detecting the production function of mitochondrial ATP ATP kit, and further application of Western The expression of Blot protein was detected by Western blotting of mitochondrial respiratory chain complex I subunit NDUFS3 protein, to explore the Drp1, effects of Marf on mitochondrial function. Results: compared with control group, the Drp1 knockdown and overexpression of Marf gene showed that Drosophila indirect flight muscle disorders, performance of flight capacity variation the abnormal rate increased, wings; electron microscope can be observed Drp1, Marf transgenic Drosophila chest muscle tissue mitochondria showed fusion, morphological damage, Drosophila indirect flight muscles are irregular, muscle fiber fracture. ATP content of mitochondrial oxidative respiration and the decrease of the Western Blot protein detection of mitochondrial respiratory complexes were significantly the content of reduced expression of I subunit of NDUFS3 protein. Conclusion: the morphological structure of Drp1 knockout and overexpression of Marf gene in Drosophila model can change the effect of the fly Drosophila mitochondria. Row function; knockout of Drp1 and overexpression of Marf gene Drosophila model can reduce mitochondrial function. It is speculated whether mitochondrial splitting or overfusion will cause mitochondrial dysfunction.

【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R741

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