Real-time monitoring of L-glutamate release from mouse brain slices under ischemia with a glass capillary-based enzyme electrode
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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 ∼10 μm containing a very small volume (∼2 μL) 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.
KeywordsGlass capillary electrode Enzyme sensor L-glutamate Mouse hippocampal slices Ischemia
The authors are grateful to T. Oka for his experimental help. This work was supported by Grants for Scientific Research by the Ministry of Education, Science and Culture, Japan. Financial support from a Research Grant from the College of Humanities and Sciences, Nihon University is also acknowledged.
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