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

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， 测量相对快速的生理代表性的第二到第二的变化，紧张浓度的 特定的神经化学物质，如血清素，在清醒的动物大脑中。微透析，a 通常使用的体内取样技术，能够测量紧张水平发生的变化。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。目前的伏安法技术可以提供 测量能力。然而，由于有限的方法， 消除背景电容电流。此外，已经显示出有限的生物结垢，限制了其用于 在清醒行为的动物中进行长期的紧张性血清素测量。所提出的电化学技术 我们称之为N形多循环方波伏安法（N-MCSWV）， 测量血清素的紧张性细胞外水平具有优异的空间分辨率，灵敏度， 特异性、选择性和减少的生物污垢。该提案利用了我们在以下方面的独特专长： 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N-MCSWV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、选择性和最小化来定量体内紧张性5-羟色胺细胞外水平 （2）将傅立叶变换电化学阻抗谱应用于N-MCSWV以监测生物污垢， 体内电极生物污染程度，和（3）验证N-MCSWV用于体内急性和慢性， 选择性测量血清素浓度。

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.