Development of an innovative in vivo voltammetric technique for measurements of tonic serotonin concentrations in the mammalian brain.

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 测量紧张剂浓度在生理上具有代表性的相对快速的秒至秒变化 清醒的动物大脑中的特定神经化学物质，如5-羟色胺。微透析，一种 通常使用的活体采样技术，能够测量发生在紧张素水平的变化。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。目前的伏安技术可以提供补药 测量能力。然而，这引起了人们对准确量化的担忧，因为对 消除本底电容电流。此外，有限的生物污垢已经被显示出来，限制了它在 清醒行为动物的长期补充性5-羟色胺测量。提出的电化学技术 我们称之为N形多循环方波伏安法(N-MCSWV)将使秒到秒 测量细胞外紧张性5-羟色胺水平，具有特殊的空间分辨率，灵敏度， 专一性、选择性和减少的生物污垢。该建议利用了我们在以下方面的独特专业知识 神经科学、电化学、软件开发和工程学来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N-MCSWV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、选择性和最小化来定量体内的紧张性5-羟色胺细胞外水平 (2)将傅里叶变换电化学阻抗谱应用于N-MCSWV的监测 以及(3)验证N-MCSWV在体内、急性和慢性的使用， 紧张性5-羟色胺浓度的选择性测量。

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 tonic concentrations of specific neurochemicals, such as serotonin, in the brains of awake behaving animals. Microdialysis, a commonly used in vivo sampling technique, is able to measure changes that occur in tonic 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. Present voltammetry technique can provide tonic measurement capability. However, it raises concerns for accurate quantification due to the limited approach to eliminate background capacitive current. Furthermore, limited biofouling has been shown, limiting its use for long-term tonic serotonin measurements in awake behaving animals. The proposed electrochemical technique we call N-shaped Multiple Cyclic Square Wave Voltammetry (N-MCSWV) will enable second-to-second measurements of tonic extracellular levels of serotonin with exceptional spatial resolution, sensitivity, specificity, selectivity, and diminished biofouling. 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 N-MCSWV as a reliable research tool that is capable of identifying and quantifying tonic serotonin extracellular levels in vivo with unsurpassed sensitivity, selectivity, and minimize biofouling, (2) apply Fourier transform electrochemical impedance spectroscopy to N-MCSWV to monitor the degree of electrode biofouling in vivo, and (3) validate the use of N-MCSWV for in vivo, acutely and chronically, selective measurement of tonic serotonin concentrations.