Glucose Oxidase-immobilized Glass Disks for Imaging of D-Glucose in Acute Brain Slices

MORIDERA Nobukatsu, YAMAMOTO Miki, OKUMURA Wataru, SUGAWARA Masao

doi:10.2116/analsci.23.39

Glucose Oxidase-immobilized Glass Disks for Imaging of D-Glucose in Acute Brain Slices

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MORIDERA Nobukatsu

Department of Chemistry, College of Humanities and Sciences, Nihon University
YAMAMOTO Miki

Department of Chemistry, College of Humanities and Sciences, Nihon University
OKUMURA Wataru

Department of Chemistry, College of Humanities and Sciences, Nihon University
SUGAWARA Masao

Department of Chemistry, College of Humanities and Sciences, Nihon University

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A biotinylated glucose oxidase (bGOD)-immobilized glass disk was prepared for visualizing D-glucose fluxes in acute brain slices. A mouse hippocampal slice was placed on the bGOD disk and stimulated with a stimulant solution containing horseradish peroxidase (HRP) and a substrate DA-64, followed by capturing digital images of Bindschedler's Green (BG), an oxidized form of DA-64, with a CCD camera. The bGOD membranes responded proportionally to D-glucose, ranging from 2.0 to 5.0 mM. Sucrose, GABA, L-glutamic acid, L-aspartic acid, glycine, acetylcholine and L-ascorbic acid at 10 mM did not cause any responses. The D-glucose fluxes in mouse hippocampal slices stimulated by a hypoxia solution were neuronal region-dependent, i.e., dentate gyrus (DG), cornu ammonis 1 (CA1) and cornu ammonis 3 (CA3), while those stimulated by KCl was independent of the neuronal regions. The response of bGOD disks is discussed in terms of the principle, concentration dependence and selectivity.