Immunocytochemical localization of alpha-D-mannosidase II in the Golgi apparatus of rat liver.
Mannosidase II is involved in the trimming of alpha-1,6-mannosyl residues during the biosynthesis of glycoproteins containing N-linked oligosaccharides of the complex type. A highly specific polyclonal antibody (IgG) was isolated from rabbits immunized with a homogeneous preparation of mannosidase II prepared from rat liver. With this antibody, light and electron microscopic immunocytochemical studies on rat liver reveal that essentially all mannosidase II in hepatocytes is localized in the Golgi apparatus, the only other site with reaction product being the endoplasmic reticulum. The indirect immunocytochemical method used in this study involved three major steps: exposure of aldehyde-fixed tissue to immune and nonimmune IgG, treatment with staphylococcal protein A labeled with horseradish peroxidase, and incubation in diaminobenzidine to reveal sites of peroxidase activity. The procedures described overcome major problems in immunocytochemistry, allowing preservation of antigenic sites and maintaining adequate ultrastructural integrity. The in situ localization of other carbohydrate-processing enzymes, involved in either trimming or attachment of sugar residues, should be possible with this procedure. Because biosynthetic precursors of the processing enzymes may be revealed by an immunocytochemical approach, it is potentially significant that mannosidase II reaction product is present in areas of the endoplasmic reticulum as well as in the Golgi apparatus.
D000818:Animals; D000936:Antigen-Antibody Complex; D006056:Golgi Apparatus / Q000201:enzymology* / Q000648:ultrastructure; D006651:Histocytochemistry; D007074:Immunoglobulin G; D008099:Liver / Q000201:enzymology* / Q000648:ultrastructure; D008361:Mannosidases / Q000378:metabolism*; D008854:Microscopy, Electron; D051381:Rats; D043323:alpha-Mannosidase
D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D008361:Mannosidases; D043323:alpha-Mannosidase