HISTOCHEMICAL DEMONSTRATION OF HUMAN LIVER HEXOKINASE USING NITRO-BLUE TETRAZOLIUM

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Histochemical demonstration of human liver hexokinase, low Km hexokinase, Km for glucose 10^<-4> M or less (HK), and glucokinase, Km for glucose10^<-1> M(GK), was described in control and liver disease patients. The method was essentially an application of kinetic assay system of HK and GK established by Vinuela et al. The activities of HK and GK were demonstrated as a dark blue deposit of formazan, which produced by the reduction of nitro blue tetrazolium(nitro-BT) with NADPH, an intermediate in the HK reaction coupled to the oxidation of glucose-6-phosphate(G6P). The specificity of present method was indicated by the lack of color development in the absence of either ATP or glucose. The intensity of formazan deposition appeared to represent the activity of HK and GK in the human liver tissue. HK was demonstrated when stained with 0.5 mM glucose concentration and HK and GK was demonstrated with 100 mM glucose concentration.
日本組織細胞化学会の論文