Regulation and function of mitochondrial motility in neurons in vivo

项目摘要 线粒体是动态细胞器，支持神经元整个生命周期的新陈代谢需求 一辈子。先前的研究试图将线粒体运输的调节定义为依赖于 神经元的活动，包括能量需求的变化。然而，之前的研究和未发表的数据 来自巴恩哈特实验室的研究表明，线粒体的运动并不依赖于神经元的活动。鉴于……的缺陷 线粒体转运与神经退行性变有关，本研究的目的是阐明 线粒体运输在维持健康和功能神经元方面的基础作用。我提议 确定跨神经元的在体线粒体定位和线粒体运动性的贡献 神经元中线粒体的功能。为了定义体内线粒体的运输和功能，我将使用 果蝇黑腹果蝇视觉系统使用遗传编码的报告进行活细胞成像 线粒体健康和神经功能。在目标1中，我将使用共聚焦显微镜、免疫染色和 EM重建确定线粒体运动性、线粒体分布之间的关系 模式和神经元结构。这一目标将评估神经元结构和 线粒体的整体定位。在目标2中，我将使用线粒体扰动来评估 在体内，线粒体的运动影响线粒体的健康和神经功能。这些研究将定义 神经元线粒体运动的调节和功能相关性，这为未来铺平了道路 研究线粒体转运缺陷在疾病中的作用，包括在神经退行性变中的作用。

Project Summary Mitochondria are dynamic organelles which support the metabolic needs of neurons across their entire lifetime. Previous research has sought to define the regulation of mitochondrial transport as dependent on the activity of the neuron, including changes in energy demand. However, previous studies and unpublished data from the Barnhart lab show mitochondrial motility is not dependent on neuronal activity. Given defects in mitochondrial transport have been implicated with neurodegeneration, the goal of this research is to elucidate the fundamental role of mitochondrial transport in maintaining healthy and functional neurons. I propose to define both in vivo mitochondrial localization across the neuron and contributions of mitochondrial motility in the function of the mitochondria in neurons. To define mitochondrial transport and function in vivo, I will use the Drosophila melanogaster visual system with live cell imaging using genetically encoded reporters of mitochondrial health and neuronal function. In Aim 1, I will use confocal microscopy, immunostaining and EM reconstructions to define the relationship between mitochondrial motility, mitochondrial distribution patterns and neuronal architecture. This aim will evaluate the relationship between neuronal structure and overall mitochondrial localization. In Aim 2, I will use mitochondrial perturbations to assess how mitochondrial motility affects mitochondrial health and neuronal function in vivo. These studies will define the regulation and functional relevance of mitochondrial motility in neurons, which paves the way towards future studies on the role of mitochondrial transport defects in disease, including in neurodegeneration.