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细胞外谷氨酸:采用微透析结合酶安培分析的在线监测

Extracellular glutamate: on-line monitoring using microdialysis coupled to enzyme-amperometric analysis.

作者信息

Zilkha E, Obrenovitch T P, Koshy A, Kusakabe H, Bennetto H P

机构信息

Gough-Cooper Department of Neurological Surgery, Institute of Neurology, London, UK.

出版信息

J Neurosci Methods. 1995 Aug;60(1-2):1-9. doi: 10.1016/0165-0270(94)00214-2.

Abstract

An enzyme-amperometric detector cell is described for flow analysis of glutamate in dialysate emerging from an implanted microdialysis probe. Its small size allows it to be placed within a few centimetres of the animal preparation, reducing the delay for data acquisition to around 2 min. The selectivity is provided by glutamate oxidase, immobilised with glutaraldehyde on surfaces adjacent to the 3-electrode system. A film of 1,2-diaminobenzene, electropolymerized on the platinum working electrode, eliminates interference from ascorbic acid and other endogenous electroactive compounds. The high sensitivity (< 0.5 mumol/l) and fast response time of the cell (90% of maximum response in 30 s) make it particularly suitable for investigating conditions that produce rapid changes in brain extracellular glutamate. This is illustrated by monitoring changes in extracellular glutamate subsequent to cardiac arrest, and K(+)-induced local depolarization.

摘要

本文描述了一种酶电流检测池,用于对植入式微透析探针流出的透析液中的谷氨酸进行流动分析。其体积小,可放置在距动物标本几厘米内,将数据采集延迟缩短至约2分钟。选择性由谷氨酸氧化酶提供,该酶用戊二醛固定在与三电极系统相邻的表面上。在铂工作电极上进行电聚合的1,2 - 二氨基苯薄膜消除了抗坏血酸和其他内源性电活性化合物的干扰。该检测池的高灵敏度(<0.5 μmol/l)和快速响应时间(30秒内达到最大响应的90%)使其特别适合研究导致脑细胞外谷氨酸快速变化的情况。通过监测心脏骤停和钾离子诱导的局部去极化后细胞外谷氨酸的变化可以说明这一点。

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