Skip to main content
SpringerLink
Log in

Role of keratan sulphate (sulphated poly -N-acetyllactosamine repeats) in keratoconic cornea, histochemical, and ultrastructural analysis

  • Cornea
  • Published:
Graefe's Archive for Clinical and Experimental Ophthalmology Aims and scope Submit manuscript

Abstract

Aims

Keratan sulphate (KS) is the predominant glycosaminoglycan (GAG) present in the corneal stroma where it is thought to regulate collagen fibril diameter. In this study we investigated the distribution of KS in normal and keratoconic corneas.

Methods

Four normal, one mild, and four severe keratoconic corneas were used for the study. Distribution of keratan sulphate proteoglycans (KS-PG) was investigated using a primary monoclonal antibody (5-D-4) that recognizes disulphated disaccharides in the poly-N-acetyllactosamine repeats of KS. The immuno-reactivity of 5-D-4 was analyzed by immunohistochemistry and immuno-electron microscopy.

Results

Immuno-histochemistry showed diffuse 5-D-4 staining in keratoconic cornea compared to the punctuate staining in normal corneas. In the single cornea with mild keratoconus, immunogold microscopy revealed a very high density of KS-PG staining, especially in the posterior stroma, compared to severe keratoconic and normal cornea. The amount of KS-PG in the stroma in severe keratoconus was slightly less compared to the normal cornea. In the mild keratoconic cornea, a higher quantity of KS-PG was present around the keratocytes. In severe keratoconic corneas, a higher quantity of KS-PG was present within the keratocytes compared to normal cornea.

Conclusions

The finding of an altered expression of KS in our keratoconic corneas, in particular the strong expression of KS in keratocytes, is in keeping with reports of an altered expression of proteoglycan metabolism in keratoconus. KS-PG plays an important role in stromal collagen fibril assembly and a dysregulation of KS-PG synthesis or catabolism could explain changes in collagen fibril spacing and diameter, which we have reported elsewhere.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Bron AJ (1988) Keratoconus. Cornea 7:163–169

    Article  PubMed  CAS  Google Scholar 

  2. Funderburgh JL, Panjwani N, Conrad GW, Baum J (1989) Altered keratan sulfate epitopes in keratoconus. Investig Ophthalmol Vis Sci 30:2278–2281

    CAS  Google Scholar 

  3. Gregory JD, Damle SP, Covington HI, Cintron C (1988) Developmental changes in proteoglycans of rabbit corneal stroma. Investig Ophthalmol Vis Sci 29:1413–1417

    CAS  Google Scholar 

  4. Hassell JR, Newsome DA, Hascall VC (1979) Characterization and biosynthesis of proteoglycans of corneal stroma from rhesus monkey. J Biol Chem 254:12346–12354

    PubMed  CAS  Google Scholar 

  5. Yue BY, Sugar J, Benveniste K (1984) Heterogeneity in keratoconus possible biochemical basis. Proc Soc Exp Biol Med 175:336–341

    PubMed  CAS  Google Scholar 

  6. Yue BY, Sugar J, Schrode K (1988) Histochemical studies of keratoconus. Curr Eye Res 7:81–86

    Article  PubMed  CAS  Google Scholar 

  7. Scott JE, Haigh M (1988) Identification of specific binding sites for keratan sulphate proteoglycans and chondroitin-dermatan sulphate proteoglycans on collagen fibrils in cornea by the use of cupromeronic blue in 'critical-electrolyte-concentration' techniques. Biochem J 253:607–610

    PubMed  CAS  Google Scholar 

  8. Chakravarti S, Magnuson T, Lass JH, Jepsen KJ, LaMantia C, Carroll H (1998) Lumican regulates collagen fibril assembly: skin fragility and corneal opacity in the absence of lumican. J Cell Biol 141:1277–1286

    Article  PubMed  CAS  Google Scholar 

  9. Bettelheim FA, Goetz D (1976) Distribution of hexosamines in bovine cornea. Investig Ophthalmol Vis Sci 15:301–304

    CAS  Google Scholar 

  10. Hassell JR, Newsome DA, Krachmer JH, Rodrigues MM (1980) Macular corneal dystrophy: failure to synthesize a mature keratan sulfate proteoglycan. Proc Natl Acad Sci USA 77:3705–3709

    Article  PubMed  CAS  Google Scholar 

  11. Nakazawa K, Hassell JR, Hascall VC, Lohmander LS, Newsome DA, Krachmer J (1984) Defective processing of keratan sulfate in macular corneal dystrophy. J Biol Chem 259:13751–13757

    PubMed  CAS  Google Scholar 

  12. Klintworth GK, Meyer R, Dennis R, Hewitt AT, Stock EL, Lenz ME, Hassell JR, Stark WJ Jr, Kuettner KE, Thonar EJ (1986) Macular corneal dystrophy. Lack of keratan sulfate in serum and cornea. Ophthalmic Paediatr Genet 7:139–143

    Article  PubMed  CAS  Google Scholar 

  13. Akama TO, Nakayama J, Nishida K, Hiraoka N, Suzuki M, McAuliffe J, Hindsgaul O, Fukuda M, Fukuda MN (2001) Human corneal GlcNac 6-O-sulfotransferase and mouse intestinal GlcNac 6-O-sulfotransferase both produce keratan sulfate. J Biol Chem 276:16271–16278

    Article  PubMed  CAS  Google Scholar 

  14. Funderburgh JL, Funderburgh ML, Rodrigues MM, Krachmer JH, Conrad GW (1990) Altered antigenicity of keratan sulfate proteoglycan in selected corneal diseases. Investig Ophthalmol Vis Sci 31:419–428

    CAS  Google Scholar 

  15. Sawaguchi S, Yue BY, Chang I, Sugar J, Robin J (1991) Proteoglycan molecules in keratoconus corneas. Investig Ophthalmol Vis Sci 32:1846–1853

    CAS  Google Scholar 

  16. Scudder P, Tang PW, Hounsell EF, Lawson AM, Mehmet H, Feizi T (1986) Isolation and characterization of sulphated oligosaccharides released from bovine corneal keratan sulphate by the action of endo-beta-galactosidase. Eur J Biochem 157:365–373

    Article  PubMed  CAS  Google Scholar 

  17. Hounsell EF, Feeney J, Scudder P, Tang PW, Feizi T (1986) 1H-NMR studies at 500 MHz of a neutral disaccharide and sulphated di-, tetra-, hexa- and larger oligosaccharides obtained by endo-beta-galactosidase treatment of keratan sulphate. Eur J Biochem 157:375–384

    Article  PubMed  CAS  Google Scholar 

  18. Mehmet H, Scudder P, Tang PW, Hounsell EF, Caterson B, Feizi T (1986) The antigenic determinants recognized by three monoclonal antibodies to keratan sulphate involve sulphated hepta- or larger oligosaccharides of the poly(N-acetyllactosamine) series. Eur J Biochem 157:385–391

    Article  PubMed  CAS  Google Scholar 

  19. Tang PW, Scudder P, Mehmet H, Hounsell EF, Feizi T (1986) Sulphate groups are involved in the antigenicity of keratan sulphate and mask i antigen expression on their poly-N-acetyllactosamine backbones. An immunochemical and chromatographic study of keratan sulphate oligosaccharides after desulphation or nitrosation. Eur J Biochem 160:537–545

    Article  PubMed  CAS  Google Scholar 

  20. Akhtar S, Bron AJ, Hawksworth NR, Bonshek RE, Meek KM (2001) Ultrastructural morphology and expression of proteoglycans, βig-h3, tenascin-C, fibrillin-1, and fibronectin in bullous keratopathy. Br J Ophthalmol 85:720–731

    Article  PubMed  CAS  Google Scholar 

  21. Kao WWY, Liu CY (2003) Role of lumican and keratocan on corneal transparency. Glycoconj J 19:275–285

    Article  Google Scholar 

  22. Sherwin T, Brookes NH, Loh IP, Poole CA, Clover GM (2002) Cellular incursion into Bowman's membrane in the peripheral cone of the keratoconus cornea. Exp Eye Res 74:473–482

    Article  PubMed  CAS  Google Scholar 

  23. Whitelock RB, Fukuchi T, Zhou L, Twining SS, Sugar J, Feder RS, Yue BY (1997) Cathepsin G, acid phosphatase, and alpha 1-proteinase inhibitor messenger RNA levels in keratoconus corneas. Investig Ophthalmol Vis Sci 38:529–534

    CAS  Google Scholar 

  24. Zhou L, Sawaguchi S, Twining SS, Sugar J, Feder RS, Yue BY (1998) Expression of degradative enzymes and protease inhibitors in corneas with keratoconus. Investig Ophthalmol Vis Sci 39:1117–11124

    CAS  Google Scholar 

  25. Kielty CM, Lees M, Shuttleworth CA, Woolley D (1993) Catabolism of intact type VI collagen microfibrils: susceptibility to degradation by serine proteinases. Biochem Biophys Res Commun 191:1230–1236

    Article  PubMed  CAS  Google Scholar 

  26. Zimmermann DR, Fischer RW, Winterhalter KH, Witmer R, Vaughan L (1988) Comparative studies of collagens in normal and keratoconus corneas. Exp Eye Res 46:431–442

    Article  PubMed  CAS  Google Scholar 

  27. Reale E, Groos S, Luciano L, Eckardt C, Eckardt U (2001) In the mammalian eye type VI collagen tetramers form three morphologically different aggregates. Matrix Biol 20:37–51

    Article  PubMed  CAS  Google Scholar 

  28. Kenney MC, Chwa M, Atilano SR, Tran A, Carballo M, Saghizadeh M, Vasiliou V, Adachi W, Brown DJ (2005) Increased levels of catalase and cathepsin V/L2 but decreased TIMP-1 in keratoconus corneas: evidence that oxidative stress plays a role in this disorder. Investig Ophthalmol Vis Sci 46:823–832

    Article  Google Scholar 

  29. Bureau J, Fabre EJ, Hecquet C, Pouliquen Y, Lorans G (1993) Modification of prostaglandin E2 and collagen synthesis in keratoconus fibroblasts, associated with an increase of interleukin 1 alpha receptor number. C R Acad Sci III 316:425–430

    PubMed  CAS  Google Scholar 

  30. Akhtar S, Bron AJ, Salvi SM, Hawksworth NR, Tuft SJ, Meek KM (2008) Collagen fibrils and proteoglycans in keratoconus - a quantitative ultra-structural analysis. Acta Ophthalmol 86:764–772

    Article  PubMed  Google Scholar 

  31. Rada JA, Cornuet PK, Hassell JR (1993) Regulation of corneal collagen fibrillogenesis in vitro by corneal proteoglycan (lumican and decorin) core proteins. Exp Eye Res 56:635–648

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Optometry, College of Applied Medical Sciences, King Saud University, PO Box 10219, Riyadh, 11433, Saudi Arabia

    S. Akhtar

  2. Nuffield Laboratory of Ophthalmology, University of Oxford, Oxford, UK

    A. J. Bron

  3. School of Biosciences, Cardiff University, Cardiff, Wales, UK

    A. J. Hayes & B. Caterson

  4. Optometry and Vision Sciences, Cardiff University, Cardiff, UK

    K. M. Meek

Authors
  1. S. Akhtar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. J. Bron
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. J. Hayes
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. K. M. Meek
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. B. Caterson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to S. Akhtar.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Akhtar, S., Bron, A.J., Hayes, A.J. et al. Role of keratan sulphate (sulphated poly -N-acetyllactosamine repeats) in keratoconic cornea, histochemical, and ultrastructural analysis. Graefes Arch Clin Exp Ophthalmol 249, 413–420 (2011). https://doi.org/10.1007/s00417-010-1512-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00417-010-1512-9

Keywords

Search

Navigation