Mechanisms and Functions of Subcellular Motility

亚细胞运动的机制和功能

基本信息

批准号：
7675326
负责人：
WILLIAM M SAXTON
金额：
$ 31.26万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA CRUZ
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-08-01 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7675326
关键词：
Alzheimer&apos s Disease Axon Axonal Transport Binding Binding Proteins Biochemical Biochemistry Biological Models Cells Collection Complex Cytoplasmic streaming Cytoskeletal Filaments Defect Dense Core Vesicle Destinations Development Dominant Genetic Conditions Dominant-Negative Mutation Drosophila genus Dynein ATPase Elements Engineering Experimental Models Filament Future Gene Mutation Genetic Genetic Models Genetic Screening Germ-Line Mutation Goals Hand Head Homologous Gene Human JNK-activating protein kinase Kinesin Knowledge Light Link MAP Kinase Kinase Kinase MAP3K1 gene MAPK8 gene Mass Spectrum Analysis Mediating Methods Microscope Microscopy Mitochondria Mitogen-Activated Protein Kinase Kinases Molecular Molecular Motors Motor Motor Neurons Movement Mutation Myosin Type V Nerve Degeneration Nervous system structure Neurodegenerative Disorders Neurons Neurotransmitters Oocytes Oogenesis Ooplasm Organelles Organism Pathway interactions Pattern Formation Peptides Phosphotransferases Process Protein Binding Proteins Regulation Research Personnel Role Scaffolding Protein Signal Pathway Staging Stream Symptoms Tail Techniques Temperature Testing Time Tissues Transgenes Transport Process Ursidae Family Walking Work cell motility digital human disease insight link protein novel novel strategies organelle movement overexpression programs retrograde transport scaffold

项目摘要

DESCRIPTION (provided by applicant): Our broad goal is to understand how cells deliver critical components to specific destinations. Such transport processes, driven by molecular motors that walk on cytoskeletal filaments, are essential in almost all cells of humans and other complex organisms. The proposed work will be done in the oocytes and neurons of Drosophila, a powerful experimental model system for studying transport and human disease mechanisms. Oocytes provide a unique opportunity to study a simplified transport mechanism. Fast streaming, a continuous movement of ooplasm, needs only one motor, kinesin-1, the motor is in a constitutively "on" state, and its linkage to cargo organelles may be relatively direct. Genetic, dominant negative, and biochemical approaches will be used to identify parts of kinesin-1 and associated proteins that link its movement to the movement of ooplasm. The results will illuminate fundamental aspects of motor- cargo linkage and help illuminate developmental pattern formation. Within neurons, results indicate that multiple types of motors cooperate in axonal transport of mitochondria and the large dense-core vesicles (DCVs) that bear peptide neurotransmitters. Time-lapse microscopy, digital organelle tracking in whole nervous systems, and powerful statistical approaches will be used in combination with genetic mutations to determine the specific sets of motors that transport mitochondria and DCVs. Novel fast-acting temperature- sensitive mutations and a fast thermal microscope stage will be engineered for rigorous tests of direct motor functions and motor-motor interactions. Also in neurons, novel regulators of mitochondrial transport will be identified and their mechanistic contributions investigated. This project uses as a springboard, our discovery that a JNK scaffolding, kinesin-binding protein (APLIP1) strongly influences retrograde mitochondrial transport. A novel genetic screen will be used to identify additional components of that control mechanism and their roles will be studied using genetics, biochemistry, and molecular approaches. Because organelle movement is such an important factor in the function and survival of neurons, defining the underlying linkage and control mechanisms is critical for understanding causes of neurodegeneration. Manipulation of transport mechanisms has great promise for future therapies that will slow the onset of the debilitating symptoms of ALS, Alzheimer's and other neurodegenerative diseases.

描述（由申请人提供）：我们的广泛目标是了解细胞如何向特定目的地传递关键组成部分。这种运输过程是由在细胞骨骼丝上行走的分子电机驱动的，几乎在人类和其他复杂生物的所有细胞中都是必不可少的。拟议的工作将在果蝇的卵母细胞和神经元中进行，果蝇是一种强大的实验模型系统，用于研究运输和人类疾病机制。卵母细胞为研究简化的运输机制提供了独特的机会。快速流媒体是卵子的连续运动，只需要一个电动机，即运动蛋白1，电动机处于组成型“状态”状态，其与货物细胞器的连锁可能相对直接。遗传，主导性阴性和生化方法将用于识别驱动蛋白-1的一部分以及将其运动与卵子运动联系起来的相关蛋白。结果将阐明汽车连锁的基本方面，并有助于阐明发展模式形成。在神经元内，结果表明，多种类型的电动机在线粒体的轴突转运和带有肽神经递质的大型密集核囊泡（DCV）中合作。延时显微镜，整个神经系统中的数字细胞器跟踪以及强大的统计方法将与基因突变结合使用，以确定传输线粒体和DCV的特定电机集。新型快速作用的温度敏感突变和快速的热显微镜阶段将被设计为直接运动功能和电动机相互作用的严格测试。同样在神经元中，将确定线粒体运输的新调节剂，并研究了其机械贡献。该项目用作跳板，这是我们发现的JNK脚手架，动力蛋白结合蛋白（APLIP1）强烈影响逆行线粒体运输。新的遗传筛查将用于鉴定该控制机制的其他组成部分及其作用，将使用遗传学，生物化学和分子方法研究。由于细胞器运动是神经元功能和存活的重要因素，因此定义潜在的联系和控制机制对于理解神经变性的原因至关重要。对运输机制的操纵对未来疗法具有巨大的希望，这些疗法将减缓ALS，阿尔茨海默氏症和其他神经退行性疾病的衰弱症状的发作。