Histochemical studies of human guanase (guanine deaminase) have seldom been undertaken, in part because of technical difficulties which result in heavy background staining. In this report, we describe a modified procedure in which the methodological inadequacies have been overcome. The modified technique has been applied to determine the intracellular and lobular distribution of guanase in normal human liver and in cases of primary biliary cirrhosis and alcoholic cirrhosis. Guanase was present within the cytoplasm of hepatocytes throughout the entire lobule. Enzyme activity was stronger on the sinusoidal side of the hepatocytes and in the periportal area. The reaction was weaker in perivenular hepatocytes. Portal components (bile ducts and veins), fibrous tissue and inflammatory cells were non-reactive, and the enzyme was absent from hepatocyte nuclei and membranes. Sections of skeletal muscle contained no guanase. The specificity of the reaction was confirmed by control tests on liver tissue and by the use of a specific inhibitor of guanase. It is concluded that the modified procedure overcomes the disadvantages inherent in the original method for guanase demonstration, allows the examination of fine cellular detail and should become a valuable histochemical tool with which to study diseases of the liver.
Human liver guanase was purified and a specific antibody against it was raised in rabbits. The antiserum formed a single precipitin line with human liver extract, and also completely inhibited the activity of the liver enzyme. An immunoblotting study showed that the antibody bound specifically to one band of protein with guanase activity and not to other proteins. Therefore, we concluded that this antiserum against the liver enzyme was suitable for use in immunohistochemical demonstration of guanase. In tissue sections, the immunohistochemical reaction with this antibody was positive in the same locations as the histochemical guanase reaction with DAB (3,3'-diaminobenzidine tetrahydrochloride).
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