Real-time monitoring of L-glutamate release from mouse brain slices under ischemia with a glass capillary-based enzyme electrode.
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.
Nakamura
Naoto
N
Department of Chemistry, College of Humanities and Sciences, Nihon University, Sakurajousui, Setagaya, Tokyo 156-8550, Japan.
Negishi
Kayoko
K
Hirano
Ayumi
A
Sugawara
Masao
M
D000594:Amino Acid Oxidoreductases / Q000378:metabolism; D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation*; D001921:Brain / Q000378:metabolism*; D005898:Glass; D018698:Glutamic Acid / Q000378:metabolism*; D007511:Ischemia / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D012680:Sensitivity and Specificity; D013302:Streptomyces; D013997:Time Factors
D018698:Glutamic Acid; D000594:Amino Acid Oxidoreductases; C023061:L-glutamate oxidase