Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Medical Center Boulevard, Winston-Salem, NC 27157, USA.
J Neurosci Methods. 2011 Nov 15;202(2):158-64. doi: 10.1016/j.jneumeth.2011.03.001. Epub 2011 Mar 8.
The fast sampling rates of fast scan cyclic voltammetry make it a favorable method for measuring changes in brain monoamine release and uptake kinetics in slice, anesthetized, and freely moving preparations. The most common analysis technique for evaluating changes in dopamine signaling uses well-established Michaelis-Menten kinetic methods that can accurately model dopamine release and uptake parameters across multiple experimental conditions. Nevertheless, over the years, many researchers have turned to other measures to estimate changes in dopamine release and uptake, yet to our knowledge no systematic comparison amongst these measures has been conducted. To address this lack of uniformity in kinetic analyses, we have created the Demon Voltammetry and Analysis software suite, which is freely available to academic and non-profit institutions. Here we present an explanation of the Demon Voltammetry acquisition and analysis features, and demonstrate its utility for acquiring voltammetric data under in vitro, in vivo anesthetized, and freely moving conditions. Additionally, the software was used to compare the sensitivity of multiple kinetic measures of release and uptake to cocaine-induced changes in electrically evoked dopamine efflux in nucleus accumbens core slices. Specifically, we examined and compared tau, full width at half height, half-life, T₂₀, T₈₀, slope, peak height, calibrated peak dopamine concentration, and area under the curve to the well-characterized Michaelis-Menten parameters, dopamine per pulse, maximal uptake rate, and apparent affinity. Based on observed results we recommend tau for measuring dopamine uptake and calibrated peak dopamine concentration for measuring dopamine release.
快速扫描循环伏安法的快速采样率使其成为测量切片、麻醉和自由活动制剂中脑单胺释放和摄取动力学变化的有利方法。评估多巴胺信号变化最常用的分析技术是使用经过充分验证的米氏-门坦动力学方法,该方法可以准确地对多种实验条件下的多巴胺释放和摄取参数进行建模。尽管如此,多年来,许多研究人员已经转向其他措施来估计多巴胺释放和摄取的变化,但据我们所知,这些措施之间没有进行过系统比较。为了解决动力学分析中缺乏一致性的问题,我们创建了 Demon Voltammetry 和 Analysis 软件套件,该套件可供学术和非营利机构免费使用。在这里,我们介绍了 Demon Voltammetry 的采集和分析功能,并展示了它在体外、体内麻醉和自由活动条件下获取伏安数据的实用性。此外,该软件还用于比较多种释放和摄取动力学测量方法对可卡因诱导的伏核核心切片中电诱发多巴胺外排变化的敏感性。具体来说,我们检查并比较了 tau、半高宽、半衰期、T₂₀、T₈₀、斜率、峰高、校准峰多巴胺浓度和曲线下面积与经过充分表征的米氏-门坦参数(多巴胺脉冲数、最大摄取率和表观亲和力)。根据观察到的结果,我们建议使用 tau 来测量多巴胺摄取,使用校准的峰多巴胺浓度来测量多巴胺释放。