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.
Moridera
Nobukatsu
N
Department of Chemistry, College of Humanities and Sciences, Nihon University, Tokyo, Japan.
Yamamoto
Miki
M
Okumura
Wataru
W
Sugawara
Masao
M
D000818:Animals; D001923:Brain Chemistry; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000302:isolation & purification; D005470:Fluorometry; D005898:Glass; D005947:Glucose / Q000032:analysis*; D005949:Glucose Oxidase / Q000737:chemistry* / Q000302:isolation & purification; D006624:Hippocampus / Q000378:metabolism; D006735:Horseradish Peroxidase; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D008297:Male; D051379:Mice
D004800:Enzymes, Immobilized; D007202:Indicators and Reagents; D005949:Glucose Oxidase; D006735:Horseradish Peroxidase; D005947:Glucose