ELECTROANALYTICAL PROBES OF EXOCYTOSIS

描述：(改编自申请人的摘要)本提案涉及 电化学检测方法的发展及应用 单次胞吐事件。拟议工作的总体目标是 进一步了解神经递质胞吐作用。这是基本的 神经递质和激素从细胞释放到 启动化学通讯。然而，仍有很多事情是未知的 胞吐作用的分子机制。申请者 建议开发制造更小电极的新策略， 可以放置在突触中，理论上应该是有用的 具有毫秒响应时间的酶电极。建议进行的实验 涉及1)调查涉及细胞控制的事件 胞吐作用的时间；2)胞吐作用外部位的调查 突触和此释放的靶受体；3)最近使用的一种 发展了胞吐作用的电化学模型，以检验 药物对神经递质浓度和囊泡大小的影响； 4)制定检查囊泡对接作用的方法学 蛋白质对胞吐释放的影响；5)制定策略 电化学测量单个突触的胞吐事件和6) 亚微米针尖直径酶电极的制备。这个 建议的工作如能顺利完成，将进一步发展 可用于测量胞吐事件的技术，并将提供 朝着理解胞吐作用的分子基础迈出了重要的一步。

DESCRIPTION: (adapted from applicant's abstract) This proposal concerns the development and application of electrochemical methods for measurements of single exocytosis events. The overall goal of the proposed work is to further understanding of neurotransmitter exocytosis. This is the basic process by which neurotransmitters and hormones are released from cells to initiate chemical communication. Yet, a great deal is still unknown about the function of the molecular mechanism of exocytosis. The applicants propose to develop new strategies for manufacturing smaller electrodes that can be placed in the synapse and should theoretically be useful to make enzyme electrodes with millisecond response time. The experiments proposed involve 1) investigation of events that involve cellular control of the timing of exocytosis; 2) investigation of sites of exocytosis outside the synapse and the target receptors for this release; 3) use of a recently developed electrochemical model of exocytosis to examine the effects of pharmacological agents on neurotransmitter concentration and vesicle size; 4) development of methodology to examine the role of vesicle-docking proteins on exocytotic release; 5) development of strategies to electrochemically measure exocytosis events in single synapses and 6) preparation of enzyme electrodes with submicron tip diameter. The successful completion of the work proposed will further develop the technology available for measurements of exocytotic events and will provide a significant step toward understanding the molecular basis of exocytosis.