Synopsis
The usefulness of a lectin,Limulus polyphemus agglutinin (LPA) has been tested in a series of mammalian tissues with sialic acid-containing glycoproteins. In nearly all the tissues employed, the positive peroxidase-labelled LPA diaminobenzidine (LPA-PO-DAB) reaction of various histological structures was markedly diminished in intensity or abolished, following digestion with neuraminidase. In the same tissues, sialic acid added with LPA-PO abolished the LPA-PO-DAB reaction or notably suppressed its intensity. In the majority of the tissues tested, the LPA-PO-DAB-Alcian Blue (AB) (pH 1.0 or 2.5) procedures appear to be useful dual staining methods which enable one to colour selectively sialic acid-containing and other acidic carbohydrates. In view of the endogenous peroxidase activity in particular histological structures, however, appropriate control staining procedures should be performed when the LPA-PO-DAB procedure is employed, either alone or in combination with AB procedures, to determine the histochemical properties of sialic acid-containing glycoproteins.
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References
Buddecke, E. (1972). Occurrence in animals and plants. In:Glycoproteins, Their Composition, Structure and Function. (ed. A. Gottschalk). pp. 535–564. Amsterdam, London, New York: Elsevier.
Faillard, H. &Schauer, R. (1972). Glycoproteins as lubricants, protective agents, carriers, structural proteins and as participants in other functions. In:Glycoproteins, Their composition, Structure and Function. (ed. A. Gottschalk). pp. 1246–1267. Amsterdam, London, New York: Elsevier.
Graham, R. C. &Karnovsky, M. B. (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.
Kienan, J. A. (1975). Localization of α-d-glucosyl and α-d-mannosyl groups of mucosubstances with concanavalin A and horseradish peroxidase.Histochemistry 44, 39–45.
Leppi, T. J. (1968). Morphochemical analysis of mucous cells in the skin and slime glands of hagfishes.Histochemie 15, 68–78.
Lev, R. &Spicer, S. S. (1964). Specific staining of sulphate groups with alcian blue at low pH.J. Histochem. Cytochem. 12, 309.
Lillie, R. D. &Fullmer, H. C. (1976).Histopathologic Technique and Practical Histochemistry. 4th Edn. pp. 25–68. New York: McGraw-Hill.
Marchalonis, J. J. &Edelman, G. M. (1968). Isolation and characterization of a hemagglutinin fromLimulus polyphemus.J. mol. Biol. 32, 453–65.
Mazzuca, M., Roche, A-C., Lhermitte, M. &Roussel, P. (1977).Limulus polyphemus lectin sites in human bronchial mucosa.J. Histochem. Cytochem. 25, 470–3.
Pearse, A. G. E. (1968).Histochemistry, Theoretical and Applied. Vol. 1. 3rd Edn. pp. 294–380. Edinburgh, London, New York: Churchill, Livingstone.
Quintarelli, G. (1961). Histochemistry of sialo-mucins in connective tissues.Histochemie 2, 356–8.
Quintarelli, G., Tsuiki, S., Hashimoto, T. &Pigman, W. (1961). Studies of sialic acid containing mucins in bovine submaxillary and rat sublingual glands.J. Histochem. Cytochem. 9, 176–83.
Roche, A-C. &Monsigny, M. (1974). Purification and properties of limulin: a lectin (agglutinin) from hemolymph ofLimulus polyphemus.Biochim. biophys. Acta 371, 242–54.
Roche, A-C., Schauer, R. &Monsigny, M. (1975). Protein-sugar interactions, purification by affinity chromatography of limulin, an N-acetyl-neuraminidyl-binding protein.FEBS Letters 57, 245–51.
Spicer, S. S. &Warren, W. (1960). The histochemistry of sialic acid containing mucoproteins.J. Histochem. Cytochem. 8, 135–7.
Spicer, S. S., Horn, R. G. &Leppi, T. J. (1967). Histochemistry of connective tissue mucopolysaccharides. In:The Connective Tissue. (eds. B. M. Wagner & D. E. Smith). pp. 251–303. Baltimore: Williams & Wilkins.
Tuppy, H. &Gottschalk, A. (1972). The structure of sialic acids and their quantitation. In:Glycoproteins, Their Composition, Structure and Function. (ed. A. Gottschalk). pp. 403–449. Amsterdam, London, New York: Elsevier.
Weir, E. E., Pretlow, T. G., Pitts, A. &Williams, E. E. (1974). Destruction of endogenous peroxidase activity in order to locate antigens by peroxidase-labelled antibodies.J. Histochem. Cytochem. 22, 51–4.
Yamada, K. (1972). Histochemical and cytochemical methods for the studies of mucosaccharides.Prot. Nucl. Acid Enzy. 17, 775–90.
Yamada, K. (1973). The effect of digestion withStreptomyces hyaluronidase upon certain histochemical reactions of hyaluronic acid-containing tissues.J. Histochem. Cytochem. 21, 794–803.
Yamada, K. (1978). Concanavalin A-peroxidase-diaminobenzidine-periodic acid-m-aminophenol-Fast Black salt K: a method for the dual staining of neutral complex carbohydrates.Histochem. J. 10, 573–84.
Yamada, K. &Ukai, M. (1976). The histochemistry of mucosacharides in some organs of germfree rats.Histochemistry 47, 219–38.
Yamada, K. &Shimizu, S. (1976). Concanavalin A-peroxidase-diaminobenzidine (Con A-PO-DAB)-alcian blue (AB). A reliable method for dual staining of complex carbohydrates.Histochemistry 47, 159–69.
Yamada, K. &Shimizu, S. (1977). The histochemistry of galactose residues of complex carbohydrates as studied by peroxidase-labelledRicinus communis agglutinin.Histochemistry 53, 143–56.
Yoshizawa, Z. (1972). Sulphated glycoproteins. In:Glycoproteins, Their Composition, Structure and Function. (ed. A. Gottschalk). pp. 1000–1018. Amsterdam, London, New York: Elsevier.
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Yamada, K., Shimizu, S. The use of peroxidase-labelledLimulus polyphemus agglutinin for the histochemistry of sialic acid-containing glycoproteins in light microscopy. Histochem J 11, 457–471 (1979). https://doi.org/10.1007/BF01002773
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DOI: https://doi.org/10.1007/BF01002773