Characterizing new genes that govern mitochondrial function in the axon

表征控制轴突线粒体功能的新基因

基本信息

批准号：
9272960
负责人：
Marc R Freeman
金额：
$ 19.25万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9272960
关键词：
Affect Amyotrophic Lateral Sclerosis Architecture Area Axon Back Biogenesis Biological Assay Biology Brain Candidate Disease Gene Cells Complex Cytoskeleton Defect Disease Disease Progression Disease model Drosophila genome Drosophila genus Functional disorder Genes Genetic Genetic Screening Health Human Image In Vitro Knowledge Lead Length Life Maintenance Mammals Measures Membrane Potentials Metabolism Mitochondria Morphology Motor Neurons Movement Mus Mutation Nerve Degeneration Neurodegenerative Disorders Neurologic Neurologic Symptoms Neurons Oxidation-Reduction Oxidative Stress PINK1 gene Parkinson Disease Pathway interactions Patients Phenotype Physiology Play Population Process Production Research Resolution Role Speed Structure Sum Superoxides System Techniques Toxin Width Work base cell motility dopaminergic neuron gene function genome-wide in vivo knock-down meter mitochondrial dysfunction mutant nervous system disorder neuronal cell body novel overexpression parkin gene/protein screening small hairpin RNA superoxide dismutase 1 tool

项目摘要

Abstract Mitochondria are integral to neuronal health. Subsequently, deficits in mitochondrial function contribute to a wealth of neurodegenerative diseases, where axonal dysfunction and die back usually precedes cell body demise. However, we know relatively little about the basic biology of mitochondrial biogenesis, morphological changes, transport, or function in axons in vivo. The discovery and characterization of new molecules regulating fundamental aspects of mitochondrial biology in axons may `open the door' to entirely new lines of research in neurodegenerative disease. In this proposal we aim to discover new regulators of mitochondria function in the axon using a novel and high throughput unbiased forward genetic screening approach recently developed in the lab. This approach allows us to assay mitochondrial morphology, number, and distribution in axons with unprecedented single axon and single mitochondrion resolution in vivo. Newly identified mitochondrial genes will then be characterized using an array of new tools we have optimized for mitochondrial studies in Drosophila, and we will determine precisely how mitochondrial physiology has been altered in vivo. We will also genetically determine how novel mitochondrial regulating genes function in defined pathways to control mitochondrial maintenance. Given that mitochondrial health and function is tightly correlated with neurodegenerative disease, it is likely that a number of these genes will play causal and/or accessory roles in neurodegeneration. We will therefore also investigate whether these novel mitochondria associated molecules have an exacerbated phenotype in dopamine neurons, since they selectively degenerate in Parkinson's disease (PD), a condition where mitochondrial dysfunction and oxidative stress is thought to play a fundamental role in disease progression. Functional conservation of these new molecules will then be assayed in mammalian neurons in vitro. This effort represents (to the best of our knowledge) the first high through forward genetic screen for molecules required for mitochondrial transport to and maintenance in axons. Thus a wealth of novel regulators of neuronal mitochondria, which have potential roles in neurological disease, await identification.

抽象的线粒体是神经元健康不可或缺的一部分。随后，线粒体功能缺陷导致大量的神经退行性疾病，轴突功能障碍和死亡通常先于细胞体出现灭亡。然而，我们对线粒体生物发生的基础生物学知之甚少，体内轴突的形态变化、运输或功能。新物质的发现和表征调节轴突线粒体生物学基本方面的分子可能“打开大门” 神经退行性疾病的新研究方向。在本提案中，我们的目标是发现新的监管机构使用新颖且高通量的无偏正向遗传筛选来研究轴突中的线粒体功能最近在实验室开发的方法。这种方法使我们能够测定线粒体形态，轴突的数量和分布具有前所未有的单轴突和单线粒体分辨率体内。然后将使用我们拥有的一系列新工具来表征新发现的线粒体基因针对果蝇线粒体研究进行了优化，我们将准确确定线粒体如何体内的生理机能发生了改变。我们还将从基因上确定新的线粒体调节方式基因在确定的途径中发挥作用来控制线粒体的维持。鉴于线粒体健康并且功能与神经退行性疾病密切相关，很可能其中许多基因将在神经变性中发挥因果和/或辅助作用。因此，我们还将调查是否这些新的线粒体相关分子在多巴胺神经元中具有加剧的表型，因为它们在帕金森病（PD）中选择性退化，这是一种线粒体功能障碍的疾病氧化应激被认为在疾病进展中发挥着重要作用。功能保护然后将在哺乳动物神经元中对这些新分子进行体外分析。这项努力代表（据我们所知）首次对所需分子进行高通量正向遗传筛选线粒体运输到轴突并在轴突中维持。因此，大量新颖的神经元调节剂线粒体在神经系统疾病中具有潜在作用，有待鉴定。