AXON-TARGETED MICRODEVICES FOR CNS AXON TRANSPORT STUDIES

用于中枢神经系统轴突运输研究的轴突靶向微器件

基本信息

批准号：
8129436
负责人：
Shelly Elese Sakiyama-Elbert
金额：
$ 22.34万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-01 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8129436
关键词：
Affect Alzheimer&apos s Disease Area Axon Axonal Transport Back Biological Assay Cell Adhesion Cell Death Cells Cellular Stress Controlled Environment Data Development Devices Dimensions Disease Disease model Environment Functional disorder Genetic Models Goals Green Fluorescent Proteins Image Impairment In Vitro Injury Knowledge Label Lead Liquid substance Microfabrication Midbrain structure Mitochondria Molecular Movement Multiple Sclerosis Mus Nerve Degeneration Nervous System Trauma Neuraxis Neurodegenerative Disorders Neurons Organelles Oxidopamine Parkinson Disease Pathology Patients Pharmaceutical Preparations Physiological Play Population Process Property Public Health Research Research Personnel Risk Role Sampling Seeds Side Signal Transduction Slide Stroke Synapses System Techniques Testing Time Toxin Transfection Trauma Tyrosine 3-Monooxygenase Work axon growth axonal degeneration design disease mechanisms study dopaminergic neuron fluorophore in vivo insight mimetics nervous system disorder neuronal cell body novel novel therapeutics promoter protein aggregation prototype public health relevance research study restraint synaptic function therapeutic development tool trafficking

项目摘要

DESCRIPTION (provided by applicant): Axonal injury is a central component in many neurological disorders including trauma, stroke, and multiple sclerosis as well as neurodegenerative disorders such as Alzheimer's (AD) and Parkinson's disease (PD). The overall goal of this research is to develop compartmentalized culture chambers using microfabrication techniques that isolate axons in culture to facilitate the study of axon growth, function and pathology. Specifically, the design and fabrication of microdevices will allow the quantitative delivery of pharmacological agents selectively to axons and allow their effects on axonal transport and growth to be examined in a controlled environment. By understanding the effects of toxins or drugs on axon growth and transport, we will gain mechanistic insight into the pathophysiology of neurodegenerative disorders and injury, which could in turn guide the development of therapeutics for treatment of degeneration or injury. The focus of this project is to develop open chamber microdevices that allow axonal isolation and the targeted application of drugs/toxins. Because patient data, genetic models, and in vivo and in vitro toxin studies all support a compelling role for axonal dysfunction in PD, we will use a well characterized PD model to study toxin effects in axons at risk in this disorder. Specifically, cultures of midbrain neurons from mice expressing green fluorescent protein (GFP) under the tyrosine hydroxylase (TH) promoter in our microdevices will allow us to test in real time the hypothesis that the Parkinson mimetic 6-OHDA (6-Hydroxydopamine) triggers changes in axonal transport resulting in the loss of axonal function and ultimately cell death. The specific aims of this proposal are to: (1) determine the design constraints for an open chamber design that allows easy cell seeding and axonal assays, which can be readily scaled for medium/high throughput cultures, (2) determine the design constraints for a novel microdevice that allows the application of a drug/toxin to axons (but not cell bodies) that are synapsing onto another population of neurons, and (3) use the new microdevices to test for the first time the effects of PD-mimetic 6-OHDA on mitochondrial trafficking in dopaminergic (DA) axons. PUBLIC HEALTH RELEVANCE: If the aims of this proposal are achieved, novel microdevices will become available to a large numbers of researchers trying to understand the role that axons play in neurological disease and injury. Establishing a causal role for axon dysfunction in neurodegenerative disorders such as Parkinson's and Alzheimer's disease is a challenging task given current methodological restraints. The tools proposed here will overcome current limitations in this field and further advance our knowledge in this area by providing unique platforms to study pharmacological and molecular mechanisms of disease and to screen for novel therapeutics.

描述（由申请人提供）：轴突损伤是许多神经系统疾病的中心成分，包括创伤，中风和多发性硬化症以及神经退行性疾病，例如阿尔茨海默氏病（AD）和帕金森氏病（PD）。这项研究的总体目标是使用微生物化技术开发分隔培养室，以分离培养物中的轴突，以促进轴突生长，功能和病理的研究。具体而言，微型版本的设计和制造将允许将药理剂定量递送至轴突，并允许在受控环境中检查其对轴突运输和生长的影响。通过了解毒素或药物对轴突生长和运输的影响，我们将获得对神经退行性疾病和损伤的病理生理学的理解，这反过来又可以指导用于治疗变性或损伤的治疗学。该项目的重点是开发允许轴突隔离和靶向使用药物/毒素的开室微发频。由于患者数据，遗传模型以及体内和体外毒素研究都支持PD中轴突功能障碍的引人注目的作用，因此我们将使用良好表征的PD模型来研究这种疾病风险的轴突中的毒素作用。 Specifically, cultures of midbrain neurons from mice expressing green fluorescent protein (GFP) under the tyrosine hydroxylase (TH) promoter in our microdevices will allow us to test in real time the hypothesis that the Parkinson mimetic 6-OHDA (6-Hydroxydopamine) triggers changes in axonal transport resulting in the loss of axonal function and ultimately cell death. The specific aims of this proposal are to: (1) determine the design constraints for an open chamber design that allows easy cell seeding and axonal assays, which can be readily scaled for medium/high throughput cultures, (2) determine the design constraints for a novel microdevice that allows the application of a drug/toxin to axons (but not cell bodies) that are synapsing onto another population of neurons, and (3) use the new microdevices to test for PD模拟6-OHDA对多巴胺能（DA）轴突线粒体运输的第一次影响。公共卫生相关性：如果实现了该提案的目的，那么大量的研究人员将可以使用新的微型版本，以了解轴突在神经系统疾病和损伤中的作用。鉴于当前的方法论限制，轴突功能障碍在神经退行性疾病（如帕金森氏病和阿尔茨海默氏病）中的因果作用是一项艰巨的任务。这里提出的工具将克服该领域的当前局限性，并通过提供独特的平台来研究疾病的药理和分子机制，并筛查新的治疗剂，从而进一步促进我们在该领域的知识。