线粒体内膜蛋白mitofilin在神经元退行性变中的作用及机制研究

项目名称： 线粒体内膜蛋白mitofilin在神经元退行性变中的作用及机制研究

项目编号： No.31471343

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 遗传学与生物信息学、细胞生物学

项目作者： 佟伟民

作者单位： 中国医学科学院基础医学研究所

项目金额： 90万元

中文摘要： 线粒体功能异常是神经元退行性病变的主要因素之一。研究表明一些神经退行性疾病中线粒体内膜蛋白mitofilin表达与转录会发生改变，但其生物学作尚不清楚。为研究mitofilin的生物学功能，我们构建了mitofilin基因敲除小鼠，初步研究结果提示mitofilin+/-小鼠呈渐进性神经退行性变。本课题将进一步探索mitofilin杂合缺失对小鼠学习、记忆、平衡等行为学的影响，通过小动物核磁、PET等脑组织分子影像分析，建立mitofilin功能缺失性神经退行性病小鼠模型。同时，通过活体荧光显微技术来研究mitofilin对不同脑区神经元线粒体功能的影响，进一步应用原代培养的神经元来研究mitofilin功能缺失对小鼠线粒体氧耗、ATP产生、膜电位、呼吸控制率、活性氧、及线粒体蛋白组成功能的等影响，探讨mitofilin在神经元退行性变发生中的作用及机制，同时探讨神经元保护性新药的作用。

中文关键词： 线粒体；神经元；mitofilin；活性氧；神经退行性病

英文摘要： Mitochondria dysfunction is one of the main factors that cause neurodegenerative diseases. It has been shown that altered expression of mitofilin, a mitochondrial inner membrane protein have been found in several neurodegenerative diseases, including Parkinson's disease. However, the role of mitofilin in the neurodegenerative diseases remain unknown. To study its biological functions in the development of neurodegenerative diseases, we generated a mitofilin-deficient mouse model using gene-trap strategy. Although mitofilin-/- mice were early embryonic lethal due to severe oxidative phosphorylation defects, preliminary study showed that mitofilin+/- mice display a progressive neurological phenotypes, resembleing neurodegenerative disorders. This project is going to investigate mice at various age groups for behavior changes, such as balance, learning and memory using ETHOVISION system and Morris water maze test, to explore the molecular imaging and morphological changes using small animal MRI, PET, and systematic pathological analysis, in order to establish a mouse model for mitofilin-deficient neurodegenerative diseases. To dissect the molecular mechanism of mitofilin in the development of neurodegenrative disorders, mitochondria function in live animals is being analyzed using in vivo fluorescence micro-image, and isolated cerebellar tissues and primary cultured cerebellar neurons are being analyzed for mitochondrial ATP production, O2 consumption, membrane potential, respiration control rate, reactive oxygen species，and mitochondrial protein constituents. Moreover, neuronal protective role of novel drugs will be tested in mitofilin-deficient mice.

英文关键词： mitochondria;neuron;mitofilin;reactive oxygen species;neurodegenerative disease