Summary
Concanavalin A is a lectin which is known to bind specifically to α-d-glucosyl and α-d-mannosyl groups in the mucosubstances of mammalian tissues. The lectin molecule is bivalent; after its attachment to mucosubstances present in a histological specimen it can also bind horseradish peroxidase, a mannose-containing glycoprotein. The attached peroxidase may then be visualized by virtue of its histochemically demonstrable enzymatic activity. Other investigators have utilized this principle in the electron microscopic localization of cell-surface carbohydrates.
A histochemical technique for light microscopy is described here, along with three control procedures which establish the specificity of the method. The technique is somewhat more fensitive than earlier ones in which fluorescent-labelled concanavalin A was used, and has the additional advantages that all the required reagents are commercially available and that sacilities for fluorescence microscopy are not needed.
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Allen, J. M., Cook, G. M. W., Poole, A. R.: Action of concanavalin A on the attachment stage of phagocytosis by macrophages. Exp. Cell Res. 68, 466–471 (1971)
Bernhard, W., Avrameas, S.: Ultrastructural visualization of cellular carbohydrate components by means of concanavalin A. Exp. Cell Res. 64, 232–236 (1971)
Cook, G. M. W., Stoddart, R. W.: Surface carbohydrates of the eukaryotic cell. London and New York: Academic Press 1974
Etzler, M. E., Branstrator, M. L.: Differential localization of cell surface and secretory components in rat intestinal epithelium. J. Cell Biol. 63, 329–343 (1974)
Goldstein, I. J., Hollerman, C. E., Smith, E. E.: Protein-carbohydrate interaction. II. Inhibition studies on the interaction of concanavalin A with polysaccharides. Biochemistry (Wash.) 4, 876–883 (1965)
Goldstein, I. J., Misaki, A.: Interaction of concanavalin A with an arabinogalactan from the cell wall of Mycobacterium bovis. J. Bact. 103, 422–425 (1970)
Graham, R. C., Karnovsky, M. B.: 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 (1966)
Heap, B. J., Kiernan, J. A.: Histological, histochemical and pharmacological observations on mast cells in the stomach of the rat. J. Anat. (Lond.) 115, 315–325 (1973)
Huet, C., Herzberg, M.: Effects of enzymes and EDTA on ruthenium red and concanavalin A labelling of the cell surface. J. Ultrastruct. Res. 42, 186–199 (1973)
Kiernan, J. A., Stoddart, R. W.: Fluorescent-labelled aprotinin: a new reagent for the histochemical detection of acid mucosubstances. Histochemie 34, 77–84 (1973)
Mallucci, L.: Binding of concanavalin A to normal and transformed cells as detected by immunofluorescence. Nature (Lond.) New Biol. 233, 241–244 (1971)
Nicolson, G. L.: Difference in topology of normal and tumour cell membranes shown by different surface distributions of ferritin-conjugated concanavalin A. Nature (Lond.) New Biol. 233, 244–245 (1971)
Nicolson, G. L., Singer, S. J.: Ferritin-conjugated plant agglutinins as specific saccharide stains for electron microscopy: application to saccharides bound to cell membranes. Proc. nat. Acad. Sci. (Wash.) 68, 942–945 (1971)
Nieland, M. L.: Epidermal intercellular staining with fluorescein-conjugated phytohemagglutinins. J. invest. Derm. 60, 61–66 (1973)
Parmley, R. J., Martin, B. J., Spicer, S. S.: Staining of blood cell surfaces with a lectinhorseradish paroxidase method. J. Histochem. Cytochem. 21, 912–922 (1973)
Pearse, A. G. E.: Histochemistry: theoretical and applied, 3rd ed., vol. 1. London: Churchill 1968
Shannon, L. M., Kay, E., Lew, J. Y.: Peroxidase isozymes from horseradish roots. I. Isolation and physical properties. J. biol. Chem. 241, 2166–2172 (1966)
Smith, C. W., Hollers, J. C.: The pattern of binding of fluorescein-labelled concanavalin A to the motile lymphocyte. J. reticuloendoth. Soc. 8, 458–464 (1970)
So, L. L., Goldstein, I. J.: Protein-carbohydrate interaction. XX. On the number of combinding sites on concanavalin A, the phytohemagglutinin of the jack bean. Biochim. biophys. Acta (Amst.) 165, 398–404 (1968)
Stoddart, R. W., Kiernan, J. A.: Histochemical detection of the α-d-arabinopyranoside configuration using fluorescent-labelled concanavalin A. Histochemie 33, 87–94 (1973)
Tkacz, J. S., Cybulska, E. B., Lampen, J. O.: Specific staining of wall mannan in yeast with fluorescein-conjugated concanavalin A. J. Bact. 106, 1–5 (1971)
Weir, E. E., Pretlow, T. G., Pitts, A., Williams, E. E.: Destruction of endogenous peroxidase activity in order to locate antigens by peroxidase-labelled antibodies. J. Histochem. Cytochem. 22, 51–54 (1974)
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Kiernan, J.A. Localization of α-d-glucosyl and α-d-mannosyl groups of mucosubstances with concanavalin A and horseradish peroxidase. Histochemistry 44, 39–45 (1975). https://doi.org/10.1007/BF00490419
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DOI: https://doi.org/10.1007/BF00490419