Development of advanced voltammetric method for basal neurotransmitter level measurement

项目总结 我们建议开发和优化一种先进的神经化学记录技术，该技术将能够 测量在生理上具有代表性的基础浓度的相对快速的秒级变化 特定的神经化学物质，如多巴胺，在清醒的行为动物的大脑中。微透析，一种 常用的活体采样技术，能够测量发生在基础水平的变化。但是，在 实践中，采样时间刻度明显限于分钟到分钟的变化，并且它的缺点是 空间分辨率，并导致严重的组织损伤。同样，传统的体内电化学 记录技术，如快速扫描循环伏安法，本质上仅限于测量相位 (刺激诱导的)神经化学浓度的变化而不是基础浓度的变化。这个 我们称之为多重循环方波伏安法(M-CSWV)的电化学技术将使 具有特殊空间的神经化学物质基础细胞外水平的秒级测量 分辨率、灵敏度、特异度和最小的组织干扰。这项建议充分利用了我们独特的专业知识 在神经科学、电化学、软件开发和工程方面开发和验证这部小说 神经化学记录技术在基础神经科学研究、临床脑中的广泛应用 神经调节，以及各种电化学应用。我们最初的动物研究将指导和告知 我们的研究技术的应用为普通神经科学和医学界所用。 我们的建议旨在(1)建立M-CSWV作为一种可靠的研究工具，能够识别和 以无与伦比的灵敏度和选择性在体内定量细胞外基础多巴胺水平；以及(2) 验证M-CSWV在体内慢性选择性测量基础多巴胺浓度和 药物诱导神经化学敏化动物模型的应用。

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.