Influence of cargo geometry on axonal transport kinetics

货物几何形状对轴突运输动力学的影响

• 批准号: 6792234
• 负责人: Sameer B. Shah
• 金额: $ 4.3万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6792234
• 关键词: amyloid proteins; axon; cell component structure /function; chemical aggregate; electron microscopy; endocytosis; fluorescent dye /probe; imaging /visualization /scanning; immunofluorescence technique; laboratory mouse; microinjections; molecular cloning; molecular size; neural degeneration; neuromuscular function; neuromuscular system; neuronal transport; neurons; postdoctoral investigator; tissue /cell culture; transfection; vesicle /vacuole

DESCRIPTION (provided by applicant): Proper function of the neuro-muscular system requires the delivery of a variety of cargoes from the neuronal cell body to destinations along the length of the axon via molecular motor-mediated transport. Given the diversity of cargoes, there is a surprising lack of information on how critical variables such as cargo size, motor density, and axonal geometry interact to affect the transport of cargo-motor complexes. The proposed experiments will test hypotheses regarding relationships between cargo size and cargo velocity, cargo run lengths, and the continuity of cargo motion. In addition, hypotheses regarding the effects of physical blockages, such as those found in the pathology of several neurodegenerative diseases, on axonal transport will be tested. Towards the testing of these hypotheses, I propose to 1) quantify the influence of cargo size on transport kinetics by imaging the motion of fluorescent beads of different diameters in cultured neurons, 2) to determine the influence of cargo size on transport kinetics by imaging the transport of vesicles engineered to specified sizes, and 3) To characterize the effects of protein accumulation on transport by introducing aggregated beta-amyloid fibrils into cultured neurons.

描述（由申请人提供）：神经肌肉系统的正确功能需要通过分子运动介导的传输将各种货物从神经元细胞体传递到沿轴突长度的目的地。考虑到货物的多样性，令人惊讶的是缺乏有关货物大小，运动密度和轴突几何形状等关键变量如何相互作用以影响货物运动配合物的运输的信息。提出的实验将检验有关货物尺寸和货物速度之间关系，货物运行长度以及货物运动连续性之间关系的假设。此外，将测试有关物理阻塞的影响的假设，例如在几种神经退行性疾病的病理中发现的假设，对轴突运输。为了测试这些假设，我建议1）通过对培养神经元中不同直径的荧光珠的运动成像，量化货物大小对传输动力学的影响，2）确定货物大小对传输动力学的影响，通过对传输动力学的影响进行成像。通过将囊泡运输到指定尺寸的囊泡的运输，以及3）通过将聚集的β-淀粉样蛋白原纤维引入培养的神经元中来表征蛋白质积累对转运的影响。