Summary
Concanavalin A lectin binding sites have been detected within the cytoplasm of epiphyseal chondrocytes. Correlative light and electron microscopic results were obtained, indicating the presence ofα-d-mannose and/or α-D-glucose residues detected by the lectin in the rough endoplasmic reticulum region. Quantitation of the electron microscopic cytochemical reaction also showed that the specific labelling was almost exclusively localized in the lumen of endoplasmic reticulum cisternae. No significant staining was found in other membrane compartments or extracellular matrix. This labelling pattern could be considered as the cytochemical evidence ofN-glycosylation processes occurring during the biosynthesis of cartilage extracellular matrix components by chondrocytes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
BAENZIGER, J. U. & FIETE, D. (1979) Structural determinants of Concanavalin A specificity for oligosaccharides.J. Biol. Chem. 254, 2400–7.
BAKER, J. R. & CATERSON, B. (1979) The isolation and characterization of the link proteins from proteoglycan aggregates of bovine nasal cartilage.J. Biol. Chem. 254, 2387–93.
CARLEMALM, E., GARAVITO, R. M. & VILLIGER, W. (1982) Resin development for electron microscopy and an analysis of embedding at low temperature.J. Microscopy 126, 123–43.
DEBRAY, H., DECOUT, D., STRECKER, G., SPIK, G. & MONTREUIL, J. (1981) Specificity of twelve lectins towards oligosaccharides and glycopeptides related toN-glycosylproteins.Eur. J. Biochem. 117, 41–55.
DE LUCA, S., LOHMANDER, L. S., NILSSON, B., HASCALL, V. C. & CAPLAN, A. I. (1980) Proteoglycans from chick limb bud chondrocyte cultures. Keratan sulfate and oligosaccharides which contain mannose and sialic acid.J. Biol. Chem. 255, 6077–83.
DORFMAN, A. (1981) Proteoglycan biosynthesis. InCell Biology of Extracellular Matrix (edited by HAY, E. D.), pp. 115–38. New York: Plenum.
FELLINI, S. A., HASCALL, V. C. & KIMURA, J. H. (1984) Localization of proteoglycan core protein in subcellular fractions isolated from rat chondrosarcoma chondrocytes.J. Biol. Chem. 259, 4634–41.
FRENS, G. (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions.Nature Phys. Sci. 241, 20–2.
GEETHA-HABIB, M., CAMPBELL, S. C. & SCHWARTZ, N. B. (1984) Subcellular localization of the synthesis and glycosylation of chondroitin sulfate proteoglycan core protein.J. Biol. Chem. 259, 7300–10.
GEOGHEGAN, W. D. & ACKERMAN, G. A. (1977) Adsorption of horseradish peroxidase, ovomucoid and antiimmunoglobulin to colloidal gold for the indirect detection of Concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level. A new method, theory and application.J. Histochem. Cytochem. 25, 1187–1200.
GOLDSTEIN, I. J., HOLLERMAN, C. E. & SMITH, E. E. (1965) Protein—carbohydrate interaction. II. Inhibition studies on the interaction of Concanavalin A with polysaccharides.Biochemistry 4, 876–83.
GRAHAM, R. C. & KARNOVSKY, M. J. (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique.J. Histochem. Cytochem. 14, 291–302.
GRIFFITHS, G., BRANDS, R., BURKE, B., LOUVARD, D. & WARREN, G. (1982) Viral membrane proteins acquire galactose in trans Golgi cisternae during intracellular transport.J. Cell Biol. 95, 781–92.
KIMURA, J. H., CAPUTO, C. B. & HASCALL, V. C. (1981) The effect of cycloheximide on synthesis of proteoglycans by cultured chondrocytes from the swarm rat chondrosarcoma.J. Biol. Chem. 256, 4368–76.
KIMURA, J. H., LOHMANDER, L. S. & HASCALL, V. C. (1984) Studies on the biosynthesis of cartilage proteoglycan in a model system of cultured chondrocytes from the swarm rat chondrosarcoma.J. Cell Biochem. 26, 261–78.
KORNFELD, R. & FERRIS, C. (1975) Interaction of immunoglobulin glycopeptides with Concanavalin A.J. Biol. Chem. 250, 2614–19.
KORNFELD, R. & KORNFELD, S. (1980) Structure of glycoproteins and their oligosaccharide units. InThe Biochemistry of Glycoproteins and Proteoglycans (edited by LENNARZ, W. J.), pp. 1–34. New York: Plenum.
LOHMANDER, L. S., DE LUCA, S., NILSSON, B., HASCALL, V. C., CAPUTO, C. B., KIMURA, J. H. & HEINEGARD, D. (1980) Oligosaccharides on proteoglycans from the swarm rat chondrosarcoma.J. Biol. Chem. 255, 6084–91.
LOHMANDER, L. S., FELLINI, S. A., KIMURA, J. H., STEVENS, R. L. & HASCALL, V. C. (1983) Formation of proteoglycan aggregates in rat chondrosarcoma chondrocyte cultures treated with tunicamycin.J. Biol. Chem. 258, 12280–6.
MITCHELL, D. & HARDINGHAM, T. (1981) The effects of cycloheximide on the biosynthesis and secretion of proteoglycans by chondrocytes in culture.Biochem. J. 196, 521–9.
NILSSON, B., DE LUCA, S., LOHMANDER, L. S. & HASCALL, V. C. (1982) Structures ofN-linked andO-linked oligosaccharides on proteoglycan monomer isolated from the swarm rat chondrosarcoma.J. Biol. Chem. 257, 10920–7.
OLSEN, B. R. (1981) Collagen biosynthesis. InCell Biology of Extracellular Matrix (edited by HAY, E. D.), pp. 139–77. New York: Plenum.
RODEN, L. (1980) Structure and metabolism of connective tissue proteoglycans. InThe Biochemistry of Glycoproteins and Proteoglycans (edited by LENNARZ, W. J.), pp. 267–371. New York: Plenum.
ROTH, J. (1983) Application of lectin—gold complexes for electron microscopic localization of glycoconjugates on thin sections.J. Histochem. Cytochem. 31, 987–99.
ROTH, J. (1984) Cytochemical localization of terminalN-acetyl-d-galactosamine residues in cellular compartments of intestinal goblet cells: implications for the topology ofO-glycosylation.J. Cell Biol. 98, 339–406.
ROUGHLEY, P. J., POOLE, A. R. & MORT, J. S. (1982) The heterogeneity of link proteins isolated from human articular cartilage proteoglycan aggregates.J. Biol. Chem. 257, 11908–14.
SCHACHTER, H. (1984) Coordination between enzyme specificity and intracellular compartmentation in the control of protein-bound oligosaccharide biosynthesis.Biol. Cell 51, 133–46.
STRUCK, D. K. & LENNARZ, W. J. (1980) The function of saccharide-lipids in synthesis of glycoproteins. InThe Biochemistry of Glycoproteins and Proteoglycans (edited by LENNARZ, W. J.), pp. 35–83. New York: Plenum.
TARTAKOFF, A. M. & VASALLI, P. (1983) Lectin-binding sites as markers of Golgi subcompartments: proximal-to-distal maturation of oligosaccharides.J. Cell Biol. 97, 1243–8.
THONAR, E. J. M. A., LOHMANDER, L. S., KIMURA, J. H., FELLINI, S. A., YANAGISHITA, M. & HASCALL, V. C. (1983) Biosynthesis ofO-linked oligosaccharides on proteoglycans by chondrocytes from the swarm rat chondrosarcoma.J. Biol. Chem. 258, 11564–70.
VERTEL, B. M., BARCKMAN, L. L. & MORRELL, J. J. (1985a) Intracellular features of type II procollagen and chondroitin sulfate proteoglycan synthesis in chondrocytes.J. Cell Biochem. 27, 215–29.
VERTEL, B. M., MORRELL, J. J. & BARCKMAN, L. L. (1985b) Immunofluorescence studies on cartilage matrix synthesis. The synthesis of link protein, chondroitin sulfate proteoglycan monomer and type II collagen.Exp. Cell Res. 158, 423–32.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Velasco, A., Hidalgo, J. Light and electron microscopical localization of concanavalin A lectin binding sites in rat epiphyseal chondrocytes. Histochem J 19, 7–14 (1987). https://doi.org/10.1007/BF01675287
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01675287