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首页> 外文期刊>Neurochemical research >A capillary fraction collector coupled to a fluorescence reader: A novel device to continuously quantify glutamate during microdialysis
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A capillary fraction collector coupled to a fluorescence reader: A novel device to continuously quantify glutamate during microdialysis

机译：与荧光读取器耦合的毛细管级分收集器：一种在微透析过程中连续定量谷氨酸的新型装置

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摘要

Microdialysis coupled to HPLC is the preferred method for quantification of glutamate (Glu) concentrations, both in normal and pathological conditions. However, HPLC is a time consuming technique that suffers from poor temporal resolution. Here we describe an alternative method to measure glutamate concentrations in small-volume dialysis samples by quantifying hydrogen peroxide released by glutamate oxidase using the Amplex Red method. This system permits continuous automatic sample collection and the detection of a fluorescent reaction product, resorufin, which provides a measure of the glutamate concentration. Quantification can be carried out in small microdialysis samples to allow a temporal resolution of 60 s. Both in vitro and in vivo tests showed that this method was reproducible and reliable, detecting Glu along a linear scale. To validate the proposed method, extracellular Glu concentrations in the rat brain were measured and correlated with electrophysiological activity prior, during and after seizure induction with 4-aminopyridine. This method may be adapted to monitor other biologically active compounds, including acetylcholine and glucose, as well as other compounds that generate hydrogen peroxide as a reaction product and may be used as an alternative to other neurochemical methods.
机译：在正常和病理条件下，微透析与HPLC相结合是定量谷氨酸（Glu）浓度的首选方法。但是，HPLC是一项耗时的技术，时间分辨率较差。在这里，我们描述了一种通过使用Amplex Red方法定量谷氨酸氧化酶释放的过氧化氢来测量小体积透析样品中谷氨酸浓度的替代方法。该系统可实现连续自动样品收集以及荧光反应产物试卤灵的检测，从而提供了谷氨酸浓度的度量​​。可以在小型微透析样品中进行定量，以实现60 s的时间分辨率。体外和体内试验均表明，该方法可重现且可靠，可线性检测Glu。为了验证所提出的方法，测量了大鼠脑中细胞外Glu的浓度，并将其与4-氨基吡啶诱发癫痫发作之前，之中和之后的电生理活性相关。该方法可适于监测其他生物活性化合物，包括乙酰胆碱和葡萄糖，以及产生过氧化氢作为反应产物的其他化合物，并可替代其他神经化学方法。

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