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Lectin Histochemistry and Cytochemistry—Light Microscopy

Avidin—Biotin Amplification on Resin-Embedded Sections

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Lectin Methods and Protocols

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 9))

Abstract

Within the last 25 yr, the study of glycans, their structures, and their drstribution in tissues has emerged from relative obscurity to become a major theme of molecular and cellular biology: “glycobiology” (1). Glycans are major components of cellular surfaces (2,3), extracellular matrices (4,5), and secretions (6), and play important roles in cell—cell and cell—matrix recognition and adhesion (7,8). They regulate the surface environment of cells by influencing the structure of water (6,9), by modulating diffusion, by sequestering metabohtes such as metal ions, by presenting various growth factors to their receptors (10,11), by acting as ligands in recognition-adhesion systems (11–13), and by making major contributions to cell surface charge (2,14). In secretions, they are variously determinants of molecular folding (15), hydration, interaction, and targeting, and they serve in the mechanisms that monitor the aging of glycoconjugates in circulation (6,9,16,17). They are now implicated in mechanisms of molecular targeting and segregation within the endomembrane systems of cells (18-20), in calcium transport in mitochondria (21), the handling of mRNA and its export from the nucleus, the regulation of transcription (19), and in then classical role as energy stores. In all of this, the association of anatomical or ultrastructural localization with biological function is of paramount importance. specific glycans occur in specific places (22,23). The challenge for the histochemist is to reconcile the achievement of sufficiently precise anatomical localization of glycans with the maximum of chemical information about their nature. The lectins have been the main means of accompllshing this, since their first application as fluorochrome-labeled probes to paraffin sections in the early 1970s (2,24). More recently, fluorescence has been largely supplanted by nonfluorescent disclosing systems, such as biotin-avidin-peroxidase and, though paraffin sections are still widely used, resin-embedding of specimens has proved advantageous in terms of resolution and economy of material at the light microscopic level.

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References

  1. Rademacher, T W, Parekh, R B, and Dwek, R A (1988) Glycobiology Ann Rev Biochem 57, 785–838

    Article  PubMed  CAS  Google Scholar 

  2. Cook, G M W and Stoddart, R W (1973) Surface Carbohydrates of the Eukaryotic Cell Academic, London, pp 346

    Google Scholar 

  3. Hughes, R C (1975) Membrane Glycoproteins A Review of Structure and Function Butterworth, London, pp 416

    Google Scholar 

  4. Kjellén, L and Lindahl, U (1991) Proteoglycans structures and interactions Ann Rev Biochem 60,443–475

    Article  PubMed  Google Scholar 

  5. Garg, H and Lyon, N (1993) Structure of collagen-fibril associated small proteoglycans of mammalian origin Adv Carbohydr Chem Biochem 49, 239–261

    Article  Google Scholar 

  6. Harding, S E (1989) The macrostructure of mucus glycoproteins in solution Adv Carbohydr Chem Biochem. 47, 345–381

    Article  PubMed  CAS  Google Scholar 

  7. Edelman, G. M. and Crossin, K. L (1991) Cell adhesion molecules implications for a molecular histology Ann Rev. Biochem 60,155–190.

    Google Scholar 

  8. Lasky, L A (1995) Selectin-carbohydrate interactions and the initiation of the inflammatory response Ann. Rev Biochem 64, 113–139

    Article  PubMed  CAS  Google Scholar 

  9. Jeffrey, G. A. (1992) Hydrogen bonding in carbohydrates and hydrate inclusion compounds Adv. Enzymol 65, 217–254

    PubMed  CAS  Google Scholar 

  10. Burgess, W H. and Macaig, T. (1989) The heparin-binding (fibroblast) growth factor family of proteins Ann Rev Biochem 58, 575–606.

    Article  PubMed  CAS  Google Scholar 

  11. Kornfeld, S (1992) Structure and function of the mannose-6-phosphate/insulinlike growth factor II receptors. Ann Rev Biochem 61, 307–330

    Article  PubMed  CAS  Google Scholar 

  12. Drickamer, K and Taylor, M. E (1993) Biology of animal lectins Ann Rev Cell Biol 9,237–264.

    Article  PubMed  CAS  Google Scholar 

  13. Turner, M L (1992) Cell adhesion molecules: a unifying approach to topographic biology Biol Rev 67, 359–377

    Article  PubMed  CAS  Google Scholar 

  14. Jeanloz, R. W and Codington, J F. (1976) The biological role of sialic acid at the surface of the cell, in Biological Roles of Sialic Acid (Rosenberg, A and Schengrund, C L, eds.), Plenum, New York, pp. 201–238

    Google Scholar 

  15. Thomas, J. (1994) Glycoproteins and glycosylation, in The Encyclopedia of Molecular Biology (Kendrew, J, ed), Blackwell Scientific, Oxford, UK, pp 439–444

    Google Scholar 

  16. Ashwell, G and Morell, A G (1974) The role of surface carbohydrates in the hepatic recognition and transport of circulating glycoproteins Adv Enzymol 41, 99–128

    PubMed  CAS  Google Scholar 

  17. Rice, K G and Lee, Y C (1993) Oligosaccharide valency and conformation in determining binding to the asialoglycoprotein receptor of rat hepatocytes Adv Enzymol 66,41–83

    PubMed  CAS  Google Scholar 

  18. Stoddart, R W (1979) Nuclear glycocoqugates and their relation to malignancy Biol Rev 54, 199–235

    Article  PubMed  CAS  Google Scholar 

  19. Hart, G W, Haltiwanger, R S, Holt, G D, and Kelly, W G (1989) Glycosylation in the nucleus and cytoplasm Ann Rev Biochem 58,841–874.

    Article  PubMed  CAS  Google Scholar 

  20. Neufeld. E F (1991) Lysosomal storage diseases Ann Rev Biochem 60,257–280

    Article  PubMed  CAS  Google Scholar 

  21. Sottocasa, G L, Sandri, G., Panfili, E, and De Bernard, B (1971) Glycoprotein located in the Inter-membrane space of rat liver mitochondria. FEBS Lett 17, 100–105

    Article  PubMed  CAS  Google Scholar 

  22. Aota, S-I and Yamada, K M (1994) Extracellular matrix molecules, in The Encyclopedia of Molecular Biology (Kendrew, J, ed.), Blackwell Science, Oxford, UK, pp 365–369

    Google Scholar 

  23. Stoddart, R W (1994) Glycans, in The Encyclopedia of Molecular Biology (Kendrew, J, ed). Blackwell Science, Oxford, UK, pp 429–436

    Google Scholar 

  24. Stoddart, R W and Kiernan, J A (1973) Histochemical detection of the α-Darabinopyranoside configuration using fluorescent-labelled Concanavalin A Histochemie 33, 87–94

    Article  PubMed  CAS  Google Scholar 

  25. Northcote, D H (1994) Lectins, in The Encyclopedia of Molecular Biology (Kendrew, J., ed.), Blackwell Science, Oxford, UK, pp 578–580

    Google Scholar 

  26. Jones, C J. P, Morrison, C A M, and Stoddart, R W (1992) Histochemical analysis of rat testicular glycoconjugates II. β-galactosyl residues in O-and N-linked glycans in semmiferous tubules Histochem J 24, 327–336.

    Article  PubMed  CAS  Google Scholar 

  27. Roberts, I S D, Jones, C J. P, and Stoddart, R W (1990) Lectin histochemistry of the mast cell heterogeneity of rodent and human mast cell populations Histochem J 22, 73–80

    Article  PubMed  CAS  Google Scholar 

  28. Bishop, P. N., Boulton, M, McLeod, D., and Stoddart, R W (1993) Glycan localization within the human interphotoreceptor matrix and photoreceptor inner and outer segments. Glycobiology 3, 403–412

    Article  PubMed  CAS  Google Scholar 

  29. Jones, C J. P, Koob, B, Stoddart, R W, Hoffman, B, and Lesser, R (1994) Lectin histochemical analysis of glycans in ovine and bovine near term binucleate cells Cell Tiss Res 278,601–610

    Article  CAS  Google Scholar 

  30. McMahon, R F T, Panesar M J R, and Stoddart, R W (1994) Glycoconjugates of the normal human colorectum. a lectin histochemical study. Histochem J 26, 504–518

    Article  PubMed  CAS  Google Scholar 

  31. Chapman, S A, Bonshek, R E., Stoddart, R W, Jones, C J P, Mackenzie, K R, O’Donoghue, E, and McLeod, D (1995) Glycoconjugates of the human trabecular meshwork a lectin histochemical study. Histochem J 27,869–881

    PubMed  CAS  Google Scholar 

  32. Robb, J L and Stoddart, R W (1988) Partial analysis of the UEA-1 binding oligosaccharide of human endothelial cells, in The Lectins (Bog-Hansen, T. and Freed, D L J, eds), Sigma, St. Louis, MO, 631–633

    Google Scholar 

  33. Jones, C J P, Kirkpatrick, C J, and Stoddart, R. W (1988) An ultrastructural study of the morphology and lectin-binding properties of human mast cell granules Histochem J 20,433–442

    Article  PubMed  CAS  Google Scholar 

  34. Sharon, N (1975) Complex Carbohydrates Their Chemistry, Biosynthesis and Functions Addison-Wesley, Reading, MA

    Google Scholar 

  35. Downs, F., Herp, A, Moscher. J, and Pigman, W (1973) β-elimination and reduction reactions and some applications of dimethylsulphoxide on submaxillary glycoproteins. Biochim Biophys Acta 328, 182–192

    PubMed  CAS  Google Scholar 

  36. Newman, G R, Jasani, B, and Williams, E D (1983) The visualisation of trace amounts of diaminobenzidine (DAB) polymer by a novel gold-sulphide-silver method J Microsc 132, RP1–RP2

    Google Scholar 

  37. Jones, C J P and Stoddart, R W (1986) A post-embedding avidin-biotin peroxidase system to demonstrate the light and electron microscopic localization of lectin binding sues in rat kidney tubules Histochem J 18,371–379

    Article  PubMed  CAS  Google Scholar 

  38. Jones, C J P, Dantzer, V, and Stoddart, R. W (1995) Changes in glycan distribution within the porcine interhaernal barrier during gestation Cell Tissue Res 279,551–564

    Article  PubMed  CAS  Google Scholar 

  39. Hsu, S-M and Raine, L (1982) Versatility of biotin-labeled lectins and avidinbiotin-peroxidase complex for localization of carbohydrate in tissue sections J Histochem Cytochem 30, 157–161

    PubMed  CAS  Google Scholar 

  40. Sarkar, M., Wu, A M., and Kabat, E. A (1981) Immunochemical studies on the carbohydrate specificity of Maclura pomifera lectin Arch Biochem Biophys 209, 204–218

    Article  PubMed  CAS  Google Scholar 

  41. Englund, P T. (1993) The structure and syntheses of glycosyl phosphatidylinosiiol protein anchors Ann Rev Biochem 62, 121–138

    Article  PubMed  CAS  Google Scholar 

  42. Jones, C J P, Aplin, J D, Mulholland, J, and Glasser, S R (1993) Patterns of sialylation in differentrating rat decidual cells as revealed by lectin histochemistry J Reprod Fert 99, 635–645

    Article  CAS  Google Scholar 

  43. Chapman, S A., Bonshek, R. E., Stoddart, R W., Mackenzie, K, and McLeod, D. (1994) Localisation of α(2,3) and α(2,6) linked terminal sialic acid groups in human trabecular meshwork Br.I. Ophthalmol 78, 632–637

    Article  CAS  Google Scholar 

  44. Jones, C. J. P, Mosley, S. M, Jeffrey, I J M, and Stoddart, R. W (1986) Elimination of the non-specific binding of avidin to tissue sections. Histochem J 19, 264–268

    Article  Google Scholar 

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Authors and Affiliations

  1. Department of Pathological Sciences, Stopford Building, University of Manchester, UK

    Robert W. Stoddart & Carolyn J. P. Jones

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  1. Robert W. Stoddart
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  2. Carolyn J. P. Jones
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Editors and Affiliations

  1. University of Liverpool, UK

    Jonathan M. Rhodes  & Jeremy D. Milton  & 

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© 1998 Humana Press Inc.

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Stoddart, R.W., Jones, C.J.P. (1998). Lectin Histochemistry and Cytochemistry—Light Microscopy. In: Rhodes, J.M., Milton, J.D. (eds) Lectin Methods and Protocols. Methods in Molecular Medicine™, vol 9. Humana Press. https://doi.org/10.1385/0-89603-396-1:21

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  • DOI: https://doi.org/10.1385/0-89603-396-1:21

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-396-2

  • Online ISBN: 978-1-59259-593-8

  • eBook Packages: Springer Protocols

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