Glucose oxidase-immobilized glass disks for imaging of D-glucose in acute brain slices.

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.