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.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Corfield AP, Carroll D, Myerscough N, Probert CS, Mucins in the gastrointestinal tract in health and disease, Frontiers in Bioscience 6, D1321–57 (2001).
van Klinken BJW, Dekker J, Buller HA, Einerhand AWC, Mucin gene structure and expression: Protection vs. adhesion, Amer. J. Physiol. 269, G613–27 (1995).
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).
Brockhausen I, Pathways of O-glycan biosynthesis in cancer cells, Biochim Biophys Acta 1473, 67–95 (1999).
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.
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).
Angata T, Varki A, Chemical diversity in the sialic acids and related keto acids. An evolutionary perspective, Chem Rev. 102, 439–69 (2002).
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.
Varki A, Sialic acids as ligands in recognition phenomena (review), FASEB J. 11, 248–55 (1997).
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).
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).
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).
Kniep B, Flegel WA, Northoff H, Rieber EP, CDw60 glycolipid antigens of human lymphocytes: Structural characterization and cellular distribution, Blood 82, 1776–86 (1993).
Reivinen H, Holthofer, Miettinen A, O-Acetylated G D3 ganglioside in human peripheral blood T lymphocytes. Int. Immunology 6, 1409–16 (1994).
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).
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).
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).
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).
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).
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).
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).
Kaswan RL, Salisbury MA, A new perspective on canine keratoconjunctivitis sicca. Treatment with ophthalmic cyclosporine, Veterinary Clinics of North America 20, 583–613 (1990).
Grauwels MFM, A study of keratoconjunctivitis sicca in the dog, PhD thesis, University of Glasgow, UK (1979).
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).
Sansom J, Barnett KC, Keratoconjunctivitis sicca in the dog: A review of two hundred cases, J Small Anim Pract 26, 121–31 (1985).
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).
Berry M, Ellingham RB, Corfield AP, Polydispersity of human ocular mucins, Invest. Ophthalmol. Vis. Sci. 37, 2559–71 (1996)
Ellingham RB, Berry M, Stevenson D, Corfield, AP, Secreted human conjunctival mucus contains MUC5AC glycoforms, Glycobiology 9, 1181–9 (1999).
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).
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).
Pearce AGE, Histochemistry, Theoretical and Applied., 3rd ed. (Churchill Livingstone, Edinburgh, 1985).
Spicer SS, Diamine methods for differentiating mucopolysaccharides histochemically, J. Histochem. Cytochem. 13, 211–34 (1965).
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).
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.
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).
Reuter G, Schauer R, Determination of sialic acids, Methods Enzymol. 230, 168–99 (1994).
Mawhinney TP, Chance DL, Hydrolysis of sialic acids and O-acetyl sialic acids with propionic acid, Anal. Biochem. 223, 164–7 (1994).
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).
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).
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).
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).
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).
Adams G, Dilly PN, Differential staining of ocular goblet cells, Eye 3, 840–4 (1989).
Ralph RA, Conjunctival goblet cell density in normal and dry eye syndromes, Invest. Ophthalmol. 14, 299–302 (1975).
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.
Holden BA, Tan KO, Sack RA, The closed-eye challenge, Adv. Exp. Med. Biol. 350, 427–30 (1994).
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).
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).
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.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Corfield, A.P., Donapaty, S.R., Carrington, S.D. et al. Identification of 9-O-acetyl-N-acetylneuraminic acid in normal canine pre-ocular tear film secreted mucins and its depletion in Keratoconjunctivitis sicca . Glycoconj J 22, 409–416 (2005). https://doi.org/10.1007/s10719-005-3698-3
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10719-005-3698-3