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Glycoconjugate Journal

, Volume 22, Issue 7–9, pp 409–416 | Cite as

Identification of 9-O-acetyl-N-acetylneuraminic acid in normal canine pre-ocular tear film secreted mucins and its depletion in Keratoconjunctivitis sicca

  • A. P. Corfield
  • S. R. Donapaty
  • S. D. Carrington
  • S. J. Hicks
  • R. Schauer
  • G. Kohla
Article

Abstract

O-Acetylated sialic acids have been reported in many sialoglycoproteins where they mediate a variety of immune and other biological events. We have previously demonstrated that the protective mucus barrier on the surface of the canine eye contains sialoglycoproteins. We have also investigated the occurrence of O-Acetylated sialic acids in these ocular mucins. Mucus aspirated from the surface of normal dog eyes and those with keratoconjunctivitis sicca (KCS) was fractionated into three pools by density gradient centrifugation. Sialic acids comprised 0.6–0.9% of the dry weight of the mucins isolated. The sialic acid profile in these pools was examined using HPLC. O-Acetylated sialic acids, mainly Neu5,9Ac2, were detected in normal animals and made up 10–30% of the total sialic acids detected. A doubling of the sialic acid content was found in KCS mucins, but the level of 9-O-Acetylated sialic acid was reduced below 4% of total.

Histological analysis of conjunctival tissue from normal and KCS dogs showed the presence of sialic acids, detected with the α(2–6) sialic acid-specific lectin Sambucus nigra, in the goblet cells and corresponding to the staining pattern for MUC5AC, the major ocular-secreted mucin gene product. In KCS animals a disruption of the normal pattern of conjunctival goblet cells was seen with preservation of the pattern of lectin binding observed in normal animals.

Thus the data demonstrate the presence of mono-O-Acetylated sialic acids in normal canine ocular mucins and a loss of this population of sialic acids in dry eye disease in spite of a significant increase in total sialic acids in KCS mucin.

Keywords

mucins sialic acids pre-ocular tear film 9-O-acetylation Keratoconjunctivitis Sicca 

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References

  1. 1.
    Corfield AP, Carroll D, Myerscough N, Probert CS, Mucins in the gastrointestinal tract in health and disease, Frontiers in Bioscience 6, D1321–57 (2001).PubMedGoogle Scholar
  2. 2.
    van Klinken BJW, Dekker J, Buller HA, Einerhand AWC, Mucin gene structure and expression: Protection vs. adhesion, Amer. J. Physiol. 269, G613–27 (1995).PubMedGoogle Scholar
  3. 3.
    Moniaux N, Escande F, Porchet N, Aubert J-P, Batra SK, Structural organisation and classification of the human mucin genes, Frontiers in Bioscience 6, 1192–1206 (2001).Google Scholar
  4. 4.
    Brockhausen I, Pathways of O-glycan biosynthesis in cancer cells, Biochim Biophys Acta 1473, 67–95 (1999).PubMedGoogle Scholar
  5. 5.
    Roussel P, Lamblin G, Human mucosal mucins in diseases, In Glycoproteins and Disease edited by Montreuil J, Vliegenthart JFG, Schachter H (Elsevier Science BV, Amsterdam, 1996), pp. 351–393.Google Scholar
  6. 6.
    Van den Steen P, Rudd PM, Dwek RA, Opdenakker G, Concepts and principles of O-linked glycosylation, Crit Rev Biochem Mol Biol 33, 151–208 (1998).PubMedGoogle Scholar
  7. 7.
    Angata T, Varki A, Chemical diversity in the sialic acids and related keto acids. An evolutionary perspective, Chem Rev. 102, 439–69 (2002).PubMedCrossRefGoogle Scholar
  8. 8.
    Schauer R, Kelm S, Reuter G, Roggentin P, Shaw L, Biochemistry and role of sialic acids., In Biology of the Sialic Acids. edited by A. Rosenberg (Plenum Press, New York, 1995), pp. 7–67.Google Scholar
  9. 9.
    Varki A, Sialic acids as ligands in recognition phenomena (review), FASEB J. 11, 248–55 (1997).PubMedGoogle Scholar
  10. 10.
    Klein, A, Krishna M, Varki NM, Varki A, 9-O-Acetylated sialic acids have widespread but selective expression: Analysis using a chimeric dual-function probe derived from influenza C hemagglutinin-esterase, Proc. Natl. Acad Sci USA 91, 7782–6 (1994).Google Scholar
  11. 11.
    Corfield AP, Myerscough N, Warren BF, Durdey P, Paraskeva C, Schauer R, Reduction of sialic acid O-acetylation in human colonic mucins in the adenoma-carcinoma sequence, Glycoconj J 16, 307–17 (1999).PubMedCrossRefGoogle Scholar
  12. 12.
    Varki A, Kornfeld S, An autosomal dominant gene regulates the extent of 9-O-acetylation of murine erythrocyte sialic acids. A probable explanation for the variation in capacity to activate the human alternate complement pathway, J. Exp. Med. 152, 532–44 (1980).PubMedCrossRefGoogle Scholar
  13. 13.
    Kniep B, Flegel WA, Northoff H, Rieber EP, CDw60 glycolipid antigens of human lymphocytes: Structural characterization and cellular distribution, Blood 82, 1776–86 (1993).PubMedGoogle Scholar
  14. 14.
    Reivinen H, Holthofer, Miettinen A, O-Acetylated GD3 ganglioside in human peripheral blood T lymphocytes. Int. Immunology 6, 1409–16 (1994).Google Scholar
  15. 15.
    Kelm S, Schauer R, Manuguerra JC, Gross HJ, Crocker PR, Modifications of cell surface sialic acids modulate cell adhesion mediated by sialoadhesin and CD22, Glycoconj. J. 11, 541–9 (1994).CrossRefGoogle Scholar
  16. 16.
    Sjoberg ER, Powell LD, Klein A, Varki A, Natural ligands of the B-cell adhesion molecule CD22β can be masked by 9-O-acetylation of sialic acids, J. Cell Biol. 126, 549–62 (1994).PubMedCrossRefGoogle Scholar
  17. 17.
    Chava AK, Bandyopadhyay S, Chatterjee M, Mandal C, Sialoglycans in protozoal diseases: Their detection, modes of aquisition and emerging biological roles, Glycoconjugate Journal 20, 199–206 (2004).PubMedGoogle Scholar
  18. 18.
    Pal S, Chatterjee M, Bhattacharya DK, Bandhyopadhyay S, Mandal C, Identification and purification of cytolytic antibodies directed against O-Acetylated sialic acid in childhood acute lymphoblastic leukemia, Glycobiology 10, 539–49 (2000).PubMedCrossRefGoogle Scholar
  19. 19.
    Harms G, Reuter G, Corfield AP, Schauer R, Binding specificity of influenza C virus to variably O-acetylated glycoconjugates and its uses for the histochemical detection of N-acetyl-9-O-acetylneuraminic acid in mammalian tissues, Glycoconjugate J. 13, 621–30 (1996).CrossRefGoogle Scholar
  20. 20.
    Klotz FW, Orlandi PA, Reuter G, Cohen SJ, Haynes JD, Schauer R, Howard RJ, Palese P, Miller LH, Binding of Plasmodium falciparum 175 kilodalton erythrocyte binding antigen and invasion of murine erythrocytes requires N-acetylneuraminic acid but not its O-Acetylated form, Mol. Biochem. Parasitol. 51, 49–54 (1992).PubMedCrossRefGoogle Scholar
  21. 21.
    Corfield AP, Carrington SD, Hicks SJ, Berry M, Ellingham R, Ocular mucins: Purification, metabolism and functions, Progress in Retinal and Eye Research 16, 627–56 (1997).CrossRefGoogle Scholar
  22. 22.
    Kaswan RL, Salisbury MA, A new perspective on canine keratoconjunctivitis sicca. Treatment with ophthalmic cyclosporine, Veterinary Clinics of North America 20, 583–613 (1990).PubMedGoogle Scholar
  23. 23.
    Grauwels MFM, A study of keratoconjunctivitis sicca in the dog, PhD thesis, University of Glasgow, UK (1979).Google Scholar
  24. 24.
    Moore, C. P., J. B. McHugh, J. G. Thorne, and T. E. Phillips. Effect of cyclosporine on conjunctival mucin in a canine keratoconjunctivitis sicca model, Invest Ophthalmol Vis Sci 42, 653–9 (2001).PubMedGoogle Scholar
  25. 25.
    Sansom J, Barnett KC, Keratoconjunctivitis sicca in the dog: A review of two hundred cases, J Small Anim Pract 26, 121–31 (1985).CrossRefGoogle Scholar
  26. 26.
    Hicks SJ, Corfield AP, Kaswan RL, Hirsh S, Stern M, Bara J, Carrington SD, Biochemical analysis of ocular surface mucin abnormalities in dry eye: the canine model, Exp Eye Res 67, 709–18 (1998).PubMedCrossRefGoogle Scholar
  27. 27.
    Berry M, Ellingham RB, Corfield AP, Polydispersity of human ocular mucins, Invest. Ophthalmol. Vis. Sci. 37, 2559–71 (1996)PubMedGoogle Scholar
  28. 28.
    Ellingham RB, Berry M, Stevenson D, Corfield, AP, Secreted human conjunctival mucus contains MUC5AC glycoforms, Glycobiology 9, 1181–9 (1999).PubMedGoogle Scholar
  29. 29.
    Carrington SD, Hicks SJ, Corfield AP, Kaswan RL, Packer N, Bolis S, Morris CA, Structural analysis of secreted ocular mucins in canine dry eye, Adv Exp Med Biol 438, 253–63 (1998).PubMedGoogle Scholar
  30. 30.
    Corfield AP, do Amaral Corfield C, Veh RW, Wagner SA, Clamp JR, Schauer R, Characterization of the major and minor mucus glycoproteins from bovine submandibular gland, Glycoconj. J. 8, 330–9 (1991).PubMedCrossRefGoogle Scholar
  31. 31.
    Pearce AGE, Histochemistry, Theoretical and Applied., 3rd ed. (Churchill Livingstone, Edinburgh, 1985).Google Scholar
  32. 32.
    Spicer SS, Diamine methods for differentiating mucopolysaccharides histochemically, J. Histochem. Cytochem. 13, 211–34 (1965).PubMedGoogle Scholar
  33. 33.
    Roe R, Corfield AP, Williamson RCN, Sialic acid in colonic mucin: An evaluation of modified PAS reactions in single and combination histochemical procedures, Histochem. J. 21, 216–22 (1989).PubMedCrossRefGoogle Scholar
  34. 34.
    Veh RW, Meessen D, Kuntz D, May B, Histochemical demonstration of side-chain substituted sialic acids., In Colon carcinogenesis, edited by Malt RA, Williamson RCN (MTP Press, Lancaster, UK, 1982), pp. 355–365.Google Scholar
  35. 35.
    Paulsen F, Thale A, Kohla G, Schauer R, Rochels R, Parwaresch R, Tillmann B, Functional anatomy of human lacrimal duct epithelium, Anat Embryol (Berl) 198, 1–12 (1998).Google Scholar
  36. 36.
    Reuter G, Schauer R, Determination of sialic acids, Methods Enzymol. 230, 168–99 (1994).PubMedGoogle Scholar
  37. 37.
    Mawhinney TP, Chance DL, Hydrolysis of sialic acids and O-acetyl sialic acids with propionic acid, Anal. Biochem. 223, 164–7 (1994).PubMedCrossRefGoogle Scholar
  38. 38.
    Hara S, Yamaguchi M, Takemori Y, Furuhata K, Ogura H, Nakamura M, Determination of mono-O-Acetylated N-acylneuraminic acids in human and rat sera by fluorimetric high performance liquid chromatography, Anal. Biochem. 179, 162–6 (1989).PubMedCrossRefGoogle Scholar
  39. 39.
    Hicks SJ, Carrington SD, Kaswan RL, Adam S, Bara J, Corfield AP, Demonstration of discrete secreted and membrane-bound ocular mucins in the dog, Experimental Eye Research 64, 597–607 (1997).PubMedCrossRefGoogle Scholar
  40. 40.
    Garcher C, Bara J, Bron A, Oriol R, Expression of mucin peptide and blood group ABH and Lewis-related carbohydrate antigens in normal human conjunctiva, Invest. Ophthalmol. Vis. Sci. 35, 1184–91 (1994).PubMedGoogle Scholar
  41. 41.
    Hazlett LD, Moon MM, Berk RS, In vivo identification of sialic acid as the ocular receptor for Pseudomonas aeruginosa, Infect. Immun. 51, 687–9 (1986).PubMedGoogle Scholar
  42. 42.
    Hazlett L, Rudner XW, Masininck S, Ireland M, Gupta S, In the immature mouse, Pseudomonas aeruginosa pili bind a 57-kD (alpha 2–6) sialylated corneal epithelial cell surface protein: a first step in infection, Invest. Ophthalmol. Vis. Sci. 36, 634–41 (1995).PubMedGoogle Scholar
  43. 43.
    Adams G, Dilly PN, Differential staining of ocular goblet cells, Eye 3, 840–4 (1989).PubMedGoogle Scholar
  44. 44.
    Ralph RA, Conjunctival goblet cell density in normal and dry eye syndromes, Invest. Ophthalmol. 14, 299–302 (1975).PubMedGoogle Scholar
  45. 45.
    Schauer R, Schmid H, Pommerencke J, Iwersen M, Kohla G, Metabolism and role of O-Acetylated sialic acids., In The Molecular Immunology of Complex Carbohydrates 2, edited by Wu AM, vol. 491. (Kluwer Academics, New York, 2001), pp. 325–342.Google Scholar
  46. 46.
    Holden BA, Tan KO, Sack RA, The closed-eye challenge, Adv. Exp. Med. Biol. 350, 427–30 (1994).PubMedGoogle Scholar
  47. 47.
    Fleiszig SMJ, Zaidi TS, Ramphal R, Pier GB, Modulation of Pseudomonas aeruginosa adherence to the corneal surface by mucus, Infect. Immun. 62, 1799–1804 (1994).PubMedGoogle Scholar
  48. 48.
    Kelm S, Schauer R, Sialic acids in Molecular and Cellular Interactions, p. 137–240. In Int. Rev. Cytology, edited by Jeon KW, Jarvik JW, vol. 175. (Academic Press, San Diego, 1997).Google Scholar
  49. 49.
    Nakamura Y, Yokoi N, Tokushige H, Kinoshita S, Sialic acid in normal and dry eye tear fluid., p. 347-. In Lacrimal Gland, Tear Film and Dry Eye Syndromes 3, edited by Sullivan D, Stern ME, Tsubota K, Dartt DA, Sullivan RM, Bromberg BB (Kluwer Academic, New York (2002), vol. 506, Part A.Google Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • A. P. Corfield
    • 1
    • 6
  • S. R. Donapaty
    • 2
  • S. D. Carrington
    • 2
    • 4
  • S. J. Hicks
    • 2
    • 5
  • R. Schauer
    • 3
  • G. Kohla
    • 3
  1. 1.Mucin Research Group, Clinical Sciences at South Bristol, Level 7Bristol Royal InfirmaryBristolUK
  2. 2.Department of Anatomy, School of Veterinary SciencesUniversity of BristolBristolUK
  3. 3.Biochemisches InstitutUniversität KielKielGermany
  4. 4.Department of Veterinary Anatomy, Faculty of Veterinary MedicineUniversity College DublinDublin 4Ireland
  5. 5.School of Applied SciencesUniversity of Wales Institute CardiffCardiffUK
  6. 6.Mucin Research Group, Clinical Sciences at South Bristol, Jenner YardBristol Royal InfirmaryBristolUK

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