Electrochemical Studies of Vesicular Release

囊泡释放的电化学研究

• 批准号: 6837284
• 负责人: CHRISTY L HAYNES
• 金额: $ 4.15万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-16 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6837284
• 关键词: acidity /alkalinity; amphetamines; calcium flux; chromaffin cells; dopamine; dopamine transporter; electrochemistry; exocytosis; fluorescence microscopy; genetically modified animals; laboratory mouse; microelectrodes; neurons; neurotransmitter transport; neurotransmitters; postdoctoral investigator; synaptic vesicles

DESCRIPTION (provided by applicant): The current understanding of neuronal exocytosis is largely based on studies done using model cell systems. Recent developments in the molecular labeling of dopaminergic neurons and microelectrode design allow single neuronal cell electroanalytical studies of dopamine release. In the proposed work, fast scan cyclic voltammetry, constant potential amperometry, and fluorescence imaging will be implemented to measure dopamine exocytosis under various conditions that affect the vesicular storage mechanisms. The frequency and amplitude of release will be monitored during the pharmacological inhibition of membrane transport machinery that regulates the entry and exit of molecules into and from the synaptic vesicle. In order to probe the role of the intravesicular matrix, weak bases will be introduced to disrupt equilibrium conditions. Manipulation of the osmolarity, pH, and temperature of the extracellular fluid will illuminate the role of solution gradients in exocytosis efficiency. This work will then extend to intact neurons from long-term culture. In effort to investigate the classical model of exocytosis, neurotransmitter release will be measured at both the cell body and synaptic terminals of neurons with intact dendrites and axons.

描述（由申请人提供）：目前对神经元胞吐作用的理解主要基于使用模型细胞系统进行的研究。多巴胺能神经元的分子标记和微电极设计的最新进展允许单神经元细胞多巴胺释放的电分析研究。在拟议的工作中，快速扫描循环伏安法，恒电位安培法，荧光成像将实施测量多巴胺胞吐在各种条件下，影响囊泡存储机制。在调节分子进出突触囊泡的膜转运机制的药理学抑制期间，将监测释放的频率和幅度。为了探索囊内基质的作用，将引入弱碱以破坏平衡条件。细胞外液的渗透压、pH和温度的操纵将阐明溶液梯度在胞吐效率中的作用。然后，这项工作将扩展到长期培养的完整神经元。为了研究胞吐作用的经典模型，将在具有完整树突和轴突的神经元的细胞体和突触末梢处测量神经递质释放。