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Immunoelectron microscopic localization of D-amino acid oxidase in rat kidney and liver
Authors:M E Perotti  E Gavazzi  L Trussardo  N Malgaretti and B Curti
Institution:(1) Department of General Physiology and Biochemistry, University of Milano, 20133 Milano, Italy;(2) Institute of Veterinary Physiology and Biochemistry, University of Milano, 20133 Milano, Italy
Abstract:Summary The intracellular localization ofd-amino acid oxidase in rat kidney and liver has been investigated using the indirect immunogold postembedding technique. Different fixation and embedding conditions for optimal preservation of antigenicity and fine structure have been tested. Immunolabelling was possible only in tissues embedded in polar resins (glycol methacrylate and Lowicryl K4M). In kidney the enzyme was demonstrable only in the peroxisomes of the proximal tubule, where it was associated with the peroxisome core. The enzyme was present in all the peroxisomes of the proximal tubule and appeared to be codistributed with catalase. Control experiments and quantitative analysis confirmed the specificity of thed-amino acid oxidase immunolocalization. All the other cells in kidney failed to demonstrate any labelling. In liver, the immunolabelling was present in the matrix of the hepatocyte peroxisomes, whereas no traces of the enzyme were found in the nucleoid. The intensity of the immunolabelling in liver peroxisomes was lower than in kidney. No specific labelling was observed in cells other than hepatocytes.
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