In vivo monitoring of cerebral agmatine by microdialysis and capillary electrophoresis
In vivo monitoring of cerebral agmatine by microdialysis and capillary electrophoresis
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DOI：
10.1016/j.jchromb.2011.11.016
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发表时间：
2012-01-01
影响因子：
3
通讯作者：
Hernandez, Luis
中科院分区：
文献类型：
--
作者：
Betancourt, Luis;Rada, Pedro;Hernandez, Luis
胍丁胺是大脑中一种假定的神经递质。目前的分析技术不允许检测细胞外液中的胍丁胺，因此很难研究其生理作用。然而，开发了一种体内监测大脑中胍丁胺的新方法。使用毛细管区带电泳和激光诱导荧光检测 (CZE-LIFD) 测量亚微升样品体积中的胍丁胺纳摩尔浓度。事实证明，该分析技术可检测 0.49 阿托摩尔的胍丁胺，提高了先前分析技术的灵敏度。另一方面，海马体是一个众所周知的大脑区域，它含有大量含有胍丁胺的神经元。因此，在海马中进行脑内微透析，并从细胞外环境中提取胍丁胺。在位于海马体的探针的透析液中发现了可检测量的胍丁胺，但在位于侧脑室的探针中未发现。此外，细胞外胍丁胺是钙和脉冲依赖性的，海马神经元的去极化增加了细胞外胍丁胺浓度。这里报道的方法足够灵敏，可以研究脑胍丁胺在自由活动的动物中的生理作用。 (C) 2011 Elsevier B.V. 保留所有权利。
Agmatine is a putative neurotransmitter in the brain. Current analytical techniques do not allow the detection of agmatine in extracellular fluid, making it difficult to study its physiological role. However, a new method for in vivo monitoring agmatine in the brain was developed. Capillary zone electrophoresis and laser induced fluorescence detection (CZE-LIFD) was used to measure nanomolar concentrations of agmatine in submicroliter sample volumes. This analytical technique proved to detect 0.49 attomole of agmatine improving the sensitivity of previous analytical techniques. On the other hand, the hippocampus is a brain region well known for having a population of agmatine containing neurons. Therefore, intracerebral microdialysis was performed in the hippocampus and agmatine was extracted from the extracellular environment. Detectable amounts of agmatine were found in dialysates from probes located in the hippocampus but not from the probes located in the lateral ventricle. Furthermore, extracellular agmatine was calcium and impulse dependent and depolarization of hippocampal neurons increased extracellular agmatine concentration. The methods reported here are sensitive enough to study the physiological role of brain agmatine in freely moving animals. (C) 2011 Elsevier B.V. All rights reserved.