Determination of GABA, glutamate and carbamathione in brain microdialysis samples by capillary electrophoresis with fluorescence detection.
Determination of GABA, glutamate and carbamathione in brain microdialysis samples by capillary electrophoresis with fluorescence detection.
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采用毛细管电泳和荧光检测法测定脑微透析样品中的 GABA、谷氨酸和氨基甲硫酮。
DOI：
10.1002/elps.201000463
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发表时间：
2011
期刊：
影响因子：
2.9
通讯作者：
Lunte,CraigE
中科院分区：
文献类型：
--
作者：
Kaul,Swetha;Faiman,MorrisD;Lunte,CraigE
双硫仑被用作治疗酗酒的威慑剂已有近 60 年的历史。我们的实验室表明，双硫仑代谢物 S-(N,N-二乙基氨基甲酰基) 谷胱甘肽（氨基甲硫酮）是由双硫仑形成的，并在服用双硫仑后出现在大脑中。氨基甲硫酮不抑制乙醛脱氢酶，但已被证明是 N-甲基-D-天冬氨酸谷氨酸 (Glu) 受体的部分非竞争性抑制剂。鉴于双硫仑对可卡因依赖的明显临床疗效以及氨基甲硫酮对 N-甲基-D-天冬氨酸受体的影响，氨基甲硫酮对脑 Glu 和 γ-氨基丁酸 (GABA) 的影响需要进一步研究。描述了一种基于萘-2,3-二甲醛衍生化的 CE-LIF 方法，可同时检测脑微透析样品中的神经递质氨基酸和氨基甲硫酮。 Glu、GABA 和氨基甲硫酮的分离使用 50mmol/L 硼酸缓冲液（pH 9.6）在 75cm×50μm 内径熔融石英毛细管（60cm 有效）上在 +27.5kV 电压下进行，运行时间为 11 分钟。 Glu、GABA 和氨基甲硫酮的检测限分别为 6、10 和 15nmol/L。该方法用于监测静脉注射药物（200mg/kg）后来自伏核的脑微透析样品中的氨基甲硫酮和氨基酸神经递质，并揭示了氨基甲硫酮诱导的 GABA 和 Glu 水平的变化。该方法演示了使用微透析采样对两种氨基酸神经递质和氨基甲硫酮进行简单、快速和准确的测量，用于大脑中的体内监测。
Disulfiram has been used as a deterrent in the treatment of alcohol abuse for almost 60 years. Our laboratory has shown that a disulfiram metabolite,S‐(N,N‐diethylcarbamoyl) glutathione (carbamathione), is formed from disulfiram and appears in the brain after the administration of disulfiram. Carbamathione does not inhibit aldehyde dehydrogenase but has been shown to be a partial non‐competitive inhibitor of theN‐methyl‐D‐aspartic acid glutamate (Glu) receptor. In light of disulfiram's apparent clinical effectiveness in cocaine dependence, and carbamathione's effect on theN‐methyl‐D‐aspartic acid receptor, the effect of carbamathione on brain Glu and γ‐aminobutyric acid (GABA) needs to be further examined. A CE‐LIF method based on derivatization with napthalene‐2,3‐dicarboxyaldehyde to simultaneously detect both neurotransmitter amino acids and carbamathione in brain microdialysis samples is described. The separation of Glu, GABA and carbamathione was carried out using a 50 mmol/L boric acid buffer (pH 9.6) on a 75 cm×50 μm id fused‐silica capillary (60 cm effective) at +27.5 kV voltage with a run time of 11 min. The detection limits for Glu, GABA and carbamathione were 6, 10 and 15 nmol/L, respectively. This method was used to monitor carbamathione and the amino acid neurotransmitters in brain microdialysis samples from the nucleus accumbens after the administration of an intravenous dose of the drug (200 mg/kg) and revealed a carbamathione‐induced change in GABA and Glu levels. This method demonstrates a simple, rapid and accurate measurement of two amino acid neurotransmitters and carbamathione for in vivo monitoring in the brain using microdialysis sampling.