THE LOCALIZATION OF N-ACETYL-β-GLUCOSAMINIDASE IN TISSUES

Abstract

α-Naphthyl N-acetyl-β-glucosaminide has been prepared and used in a simultaneous azo-dye coupling method for the demonstration of N-acetyl-β-glucosaminidase activity in formalin fixed frozen sections The effect of fixation on the activity of the enzyme has been invesitgated. In addition the effects of varying the substrate concentration, amount of diazonium salt and pH of the incubating medium are described N-Acetylglucosaminonolactone, a specific inhibitor of N-acetyl-β-glucosaminidase has been used in experiments designed to test the specificity of the reaction. The relative activity of the enzyme in rat tissue homogenates has been measured and its distribution in rat tissues and human cartilage and synovial membrane is described A satisfactory localization of N-acetyl-β-glucosaminidase is possible by the technique described. Factors affecting the histochemical localization are discussed in relation to the results obtained with other enzymes