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）：当前对神经胞吐作用的理解主要基于使用模型细胞系统进行的研究。多巴胺能神经元和微电极设计的分子标记的最新发展允许多巴胺释放的单个神经元细胞电分析研究。在拟议的工作中，将实施快速扫描循环伏安法，恒定电位安培法和荧光成像，以在影响囊泡存储机制的各种条件下测量多巴胺胞吐作用。释放的频率和振幅将在对膜转运机械的药理抑制过程中进行监测，该膜转运机械调节分子进入突触囊泡的进入和退出。为了探测内部矩阵的作用，将引入弱碱基以破坏平衡条件。对细胞外液的渗透性，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文化的完整神经元。为了研究胞吐作用的经典模型，将在具有完整的树突和轴突的神经元的细胞体和突触末端测量神经递质释放。