Schier Christina J, Mangieri Regina A, Dilly Geoffrey A, Gonzales Rueben A
College of Pharmacy, Division of Pharmacology and Toxicology, The University of Texas at Austin, Austin, Texas, USA.
J Vis Exp. 2012 Sep 5(67):4142. doi: 10.3791/4142.
Operant self-administration methods are commonly used to study the behavioral and pharmacological effects of many drugs of abuse, including ethanol. However, ethanol is typically self-administered orally, rather than intravenously like many other drugs of abuse. The pharmacokinetics of orally administered drugs are more complex than intravenously administered drugs. Because understanding the relationship between the pharmacological and behavioral effects of ethanol requires knowledge of the time course of ethanol reaching the brain during and after drinking, we use in vivo microdialysis and gas chromatography with flame ionization detection to monitor brain dialysate ethanol concentrations over time. Combined microdialysis-behavioral experiments involve the use of several techniques. In this article, stereotaxic surgery, behavioral training and microdialysis, which can be adapted to test a multitude of self-administration and neurochemical centered hypotheses, are included only to illustrate how they relate to the subsequent phases of sample collection and dialysate ethanol analysis. Dialysate ethanol concentration analysis via gas chromatography with flame-ionization detection, which is specific to ethanol studies, is described in detail. Data produced by these methods reveal the pattern of ethanol reaching the brain during the self-administration procedure, and when paired with neurochemical analysis of the same dialysate samples, allows conclusions to be made regarding the pharmacological and behavioral effects of ethanol.
操作式自我给药方法常用于研究包括乙醇在内的许多滥用药物的行为和药理作用。然而,乙醇通常是口服自我给药,而不像许多其他滥用药物那样通过静脉注射。口服给药的药物药代动力学比静脉注射给药的药物更为复杂。由于理解乙醇的药理作用与行为效应之间的关系需要了解饮酒期间及之后乙醇到达大脑的时间进程,我们使用体内微透析和带有火焰离子化检测的气相色谱法来监测脑透析液中乙醇浓度随时间的变化。微透析与行为相结合的实验涉及多种技术的运用。在本文中,立体定位手术、行为训练和微透析(这些方法可用于检验众多以自我给药和神经化学为中心的假设)仅用于说明它们与后续样本采集和透析液乙醇分析阶段的关系。本文详细描述了通过具有火焰离子化检测功能的气相色谱法进行透析液乙醇浓度分析,这一方法专门用于乙醇研究。这些方法产生的数据揭示了自我给药过程中乙醇到达大脑的模式,并且当与相同透析液样本的神经化学分析相结合时,能够得出关于乙醇药理作用和行为效应的结论。