Electroanalytical Probes of Exocytosis

胞吐作用的电分析探针

• 批准号: 6331685
• 负责人: ANDREW G EWING
• 金额: $ 22.41万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6331685
• 关键词: PC12 cells; analytical method; chromatography; electrochemistry; electrophysiology; enzyme electrodes; exocytosis; fluorescent dye /probe; method development; neural plasticity; neurons; neurotransmitter receptor; neurotransmitter transport; single cell analysis; synapses; tissue /cell culture

This proposal is concerned with the development and application of methods in electrochemical extremely small environments aimed at single nerve cells. The underlying goals of the proposed work are to develop a better fundamental understanding of neurotransmission by examining exocytosis at neuronal cells bodies, mammalian cells in culture, and in single neuronal synapses. This approach combines developments in analytical measurements with experiments to investigate new aspects of neurochemistry. The methods under development use electrochemistry to probe ultrasmall environments in neuroscience. New aspects include systems to model electrochemical responses, capillary/electrochemistry measurements of vesicles, and nanometer tip electrodes for the femtoliter environment of the synapse. Neurochemical experiments are targeted at understanding exoytosis at the molecular level, discovering the neuronal rationale for neurotransmitter release at the cell body, and understanding concentration and volume effects in transmitter release during exocytosis. The later issue could be important in understanding factors involved in the treatment and/or progression of Parkinson's Disease. The specific plans of the research are: 1, to model exocytosis events measured at single electrodes; 2, to use electrochemical probes to measure plasticity mediated by the vesicular monoamine transporter; 3, to develop and apply hydrodynamic chromatography in nanometer capillaries with electrochemical detection to measure vesicle size and neurotransmitter amount for single vesicles; 4, to investigate with electrochemical and fluorescent methods the factors regulating the function of transmitter release from the neuronal cell body; and 5, to carry out electrochemical measurements in single synapses.

这一建议是有关方法的发展和应用在电化学极小的环境中针对单个神经细胞。提出的工作的潜在目标是通过检查神经元细胞体，培养的哺乳动物细胞和单个神经元突触的胞吐作用，对神经传递有更好的基础理解。这种方法将分析测量的发展与实验相结合，以研究神经化学的新方面。正在开发的方法使用电化学来探测神经科学中的超小环境。新的方面包括模拟电化学反应的系统，囊泡的毛细管/电化学测量，以及用于突触飞升环境的纳米尖端电极。神经化学实验的目标是在分子水平上理解胞吐，发现神经递质在细胞体释放的神经元原理，并了解胞吐过程中递质释放的浓度和体积效应。后一个问题对于理解帕金森病治疗和/或进展的相关因素可能很重要。本研究的具体计划是：1、模拟单电极测量的胞吐事件；2、利用电化学探针测量水疱单胺转运体介导的可塑性；3、开发并应用纳米毛细管水动力色谱电化学检测技术，测量单个微泡的大小和神经递质含量；4、用电化学和荧光法研究神经元胞体释放递质功能的调控因素；5，在单个突触中进行电化学测量。