Development of advanced voltammetric method for basal neurotransmitter level measurement

项目摘要 我们建议开发和优化一种先进的神经化学记录技术， 测量基础浓度的相对快速的生理代表性秒到秒的变化， 特定的神经化学物质，如多巴胺，在清醒的动物大脑中。微透析，a 通常使用的体内取样技术，能够测量基础水平发生的变化。但在 在实践中，采样时间尺度明显限于分钟到分钟的变化， 空间分辨率并引起显著的组织损伤。同样，常规的体内电化学 记录技术，例如快速扫描循环伏安法，本质上限于测量相位 （刺激诱导的）神经化学浓度的变化，而不是基础浓度的变化。的 我们提出的称为多循环方波伏安法（M-CSWV）的电化学技术将使 每秒测量细胞外神经化学物质的基础水平， 分辨率、灵敏度、特异性和最小的组织干扰。该提案利用了我们独特的专业知识 在神经科学，电化学，软件开发和工程开发和验证这一新的 神经化学记录技术广泛应用于基础神经科学研究、临床脑 神经调节和各种电化学应用。我们最初的动物研究将指导和告知 我们的研究技术在普通神经科学和医学界的应用。 我们的建议旨在（1）建立M-CSWV作为一个可靠的研究工具，能够识别和 以无与伦比的灵敏度和选择性定量体内基础多巴胺细胞外水平;和（2） 验证使用M-CSWV进行基础多巴胺浓度的体内慢性选择性测量， 在药物诱导的神经化学致敏的动物模型中的应用。

PROJECT SUMMARY We propose to develop and optimize an advanced neurochemical recording technique that would be able to measure relatively rapid physiologically representative second-to-second changes in basal concentrations of specific neurochemicals, such as dopamine, in the brains of awake behaving animals. Microdialysis, a commonly used in vivo sampling technique, is able to measure changes that occur in basal levels. However, in practice the sampling timescale is significantly limited to minute-to-minute changes and it suffers from poor spatial resolution and induces significant tissue damage. As well, conventional in vivo electrochemical recording techniques, such as fast-scan cyclic voltammetry, are intrinsically limited to measuring phasic (stimulation-induced) changes in neurochemical concentrations and not changes in basal concentrations. The proposed electrochemical technique we call Multiple Cyclic Square Wave Voltammetry (M-CSWV) will enable second-to-second measurements of basal extracellular levels of neurochemicals with exceptional spatial resolution, sensitivity, specificity, and minimal tissue disturbance. This proposal leverages our unique expertise in neuroscience, electrochemistry, software development, and engineering to develop and validate this novel neurochemical recording technology for broad use in basic neuroscience research, clinical brain neuromodulation, and a variety of electrochemical applications. Our initial animal studies will guide and inform the application of our investigational technique for use by the general neuroscience and medical community. Our proposal seeks to (1) establish M-CSWV as a reliable research tool that is capable of identifying and quantifying basal dopamine extracellular levels in vivo with unsurpassed sensitivity and selectivity; and (2) validate the use of M-CSWV for in vivo chronic selective measurement of basal dopamine concentrations and application in an animal model of drug-induced neurochemical sensitization.