Nakamura Naoto, Negishi Kayoko, Hirano Ayumi, Sugawara Masao
Department of Chemistry, College of Humanities and Sciences, Nihon University, Sakurajousui, Setagaya, Tokyo 156-8550, Japan.
Anal Bioanal Chem. 2005 Oct;383(4):660-7. doi: 10.1007/s00216-005-0033-6. Epub 2005 Oct 19.
Real-time monitoring of L-glutamate release from various neuronal regions of mouse hippocampal slices under ischemia (a glucose-free hypoxia condition) is described. A glass capillary microelectrode with a tip size of approximately 10 microm containing a very small volume ( approximately 2 microL) of a solution of glutamate oxidase (GluOx) and ascorbate oxidase was used. First, the amperometric response behavior of the electrode at 0 V versus Ag/AgCl was characterized with a standard glutamate solution in terms of continuous measurements, effect of oxygen, viscosity of solution and concentration dependence. The electrode was applied to the real-time monitoring of L-glutamate released from different neuronal regions of acute hippocampal slices submerged in a hypoxia solution. The time-resolved amounts of L-glutamate released at various neuronal regions (CA1, CA3 and DG) of mouse hippocampal slices were quantified and compared with the reported L-glutamate fluxes using difference-image analysis during ischemia.
本文描述了在缺血（无糖缺氧条件）下对小鼠海马切片不同神经元区域L-谷氨酸释放的实时监测。使用了一种尖端尺寸约为10微米的玻璃毛细管微电极，其中含有极少量（约2微升）的谷氨酸氧化酶（GluOx）和抗坏血酸氧化酶溶液。首先，通过连续测量、氧气影响、溶液粘度和浓度依赖性，用标准谷氨酸溶液对电极在相对于Ag/AgCl为0 V时的安培响应行为进行了表征。将该电极应用于对浸没在缺氧溶液中的急性海马切片不同神经元区域释放的L-谷氨酸的实时监测。使用差异图像分析对小鼠海马切片不同神经元区域（CA1、CA3和DG）在缺血期间释放的L-谷氨酸的时间分辨量进行了定量，并与报道的L-谷氨酸通量进行了比较。
