Skip to main content
SpringerLink
Log in

Differential immunogold localisation of sulphated and unsulphated keratan sulphate proteoglycans in normal and macular dystrophy cornea using sulphation motif-specific antibodies

  • Short Communication
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

Keratan sulphate (KS) proteoglycans (PGs) are key molecules in the corneal stroma for tissue organisation and transparency. Macular corneal dystrophy (MCD) is a rare, autosomal recessive disease characterised by disturbances in KS expression. MCD is caused by mutations in CHST6, a gene encoding the enzyme responsible for KS sulphation. Sulphated KS is absent in type I disease causing corneal opacity and loss of vision. Genetic studies have highlighted the mutational heterogeneity in MCD, but supportive immunohistochemical studies on corneal KS have previously been limited by the availability of antibodies mostly reactive only with highly sulphated KS epitopes. In this study, we employed four antibodies against specific KS sulphation patterns, including one against unsulphated KS, to investigate their reactivity in a case of MCD compared with normal cornea using high-resolution immunogold electron microscopy. Mutation analysis indicated type I MCD with deletion of the entire open reading frame of CHST6. Contrast enhanced fixation revealed larger PG structures in MCD than normal. Unlike normal cornea, MCD cornea showed positive labelling with antibody to unsulphated KSPG, but was negative with antibodies to sulphated KSPG. These antibodies will thus facilitate high-resolution investigations of phenotypic heterogeneity in support of genetic studies in this disease.

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.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Akama TO, Nishida K, Nakayama J, Watanabe H, Ozaki K, Nakamura T, Dota A, Kawasaki S, Inoue Y, Maeda N, Yamamoto S, Fujiwara T, Thonar EJ, Shimomura Y, Kinoshita S, Tanigami A, Fukuda MN (2000) Macular corneal dystrophy type I and type II are caused by distinct mutations in a new sulphotransferase gene. Nat Genet 26:237–241

    Article  PubMed  CAS  Google Scholar 

  • 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 sulphate. J Biol Chem 276:16271–16278

    Article  PubMed  CAS  Google Scholar 

  • Caterson B, Christner JE, Baker JR, Couchman JR (1985) Production and characterization of monoclonal antibodies directed against connective tissue proteoglycans. Federation Proc 44:386–393

    CAS  Google Scholar 

  • 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 

  • Edward DP, Yue BY, Sugar J, Thonar EJ, SunderRaj N, Stock EL, Tso MO (1988) Heterogeneity in macular corneal dystrophy. Arch Ophthalmol 106:1579–83

    PubMed  CAS  Google Scholar 

  • El-Ashry MF, Abd El-Aziz MM, Shalaby O, Wilkins S, Poopalasundaram S, Cheetham M, Tuft SJ, Hardcastle AJ, Bhattacharya SS, Ebenezer ND (2005) Novel CHST6 nonsense and missense mutations responsible for macular corneal dystrophy. Am J Ophthalmol 139:192–193

    Article  PubMed  CAS  Google Scholar 

  • Funderburgh JL (2000) Keratan sulfate: structure, biosynthesis, and function. Glycobiology 10:951–958

    Article  PubMed  CAS  Google Scholar 

  • Iida-Hasegawa N, Furuhata A, Hayatsu H, Murakami A, Fujiki K, Nakayasu K, Kanai A (2003) Mutations in the CHST6 gene in patients with macular corneal dystrophy: immunohistochemical evidence of heterogeneity. Invest Ophthalmol Vis Sci 44:3272–3277

    Article  PubMed  Google Scholar 

  • Kaufer JN, Hakomori S (1983) Glycolipid antigens and genetic markers. In: Hanahan DJ (eds) Handbook of lipid research—sphingolipid biochemistry. Plenum, New York, pp 409–429

    Google Scholar 

  • Klintworth GK, Oshima E, al-Rajhi A, al-Salif A, Thonar EJ, Karcioglu ZA (1997) Macular corneal dystrophy in Saudi Arabia: a study of 56 cases and recognition of a new immunophenotype. Am J Ophthalmol 124:9–18

    PubMed  CAS  Google Scholar 

  • Klintworth GK, Smith CF, Bowling BL (2006) CHST6 mutations in North American subjects with macular corneal dystrophy: a comprehensive molecular genetic review. Mol Vis 12:159–176

    PubMed  CAS  Google Scholar 

  • Lewis D, Davies Y, Nieduszynski IA, Lawrence F, Quantock AJ, Bonshek R, Fullwood NJ (2000) Ultrastructural localization of sulphated and unsulfated keratan sulphate in normal and macular corneal dystrophy type I. Glycobiology 10:305–312

    Article  PubMed  CAS  Google Scholar 

  • Meek KM, Quantock AJ, Elliott GF, Ridgway AE, Tullo AB, Bron AJ, Thonar EJ (1989) Macular corneal dystrophy: the macromolecular structure of the stroma observed using electron microscopy and synchrotron X-ray diffraction. Exp Eye Res 49:941–958

    Article  PubMed  CAS  Google Scholar 

  • 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 

  • Plaas AH, West LA, Thonar EJ, Karcioglu ZA, Smith CJ, Klintworth GK, Hascall VC (2001) Altered fine structures of corneal and skeletal keratan sulphate and chondroitin/dermatan sulphate in macular corneal dystrophy. J Biol Chem 276:39788–39796

    Article  PubMed  CAS  Google Scholar 

  • Quantock AJ, Meek KM, Thonar EJ, Assil KK (1993) Synchrotron X-ray diffraction in atypical macular dystrophy. Eye 7:779–784

    PubMed  Google Scholar 

  • Scott JE, Haigh M (1988) Identification of specific binding sites for keratan sulfate proteoglycans and chondroitin-dermatan sulfate 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 

  • Sultana A, Sridhar MS, Klintworth GK, Balasubramanian D, Kannabiran C (2005) Allelic heterogeneity of the carbohydrate sulfotransferase-6 gene in patients with macular corneal dystrophy. Clin Genet 68:454–460

    Article  PubMed  CAS  Google Scholar 

  • Warren JF, Aldave AJ, Srinivasan M, Thonar EJ, Kumar AB, Cevallos V et al (2003) Novel mutations in the CHST6 gene associated with macular corneal dystrophy in southern India. Arch Ophthalmol 121:1608–1612

    Article  PubMed  CAS  Google Scholar 

  • Yang CJ, SundarRaj N, Thonar EJ, KlintworthGK (1988) Immunohistochemical evidence of heterogeneity in macular corneal dystrophy. Am J Ophthalmol 106:65–71

    PubMed  CAS  Google Scholar 

  • Young RD, Tudor D, Hayes AJ, Kerr B, Hayashida Y, Nishida K, Meek KM, Caterson B, Quantock AJ (2005) Atypical composition and ultrastructure of proteoglycans in the mouse corneal stroma. Invest Ophthalmol Vis Sci 46:1973–1978

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Structural Biophysics Group, School of Optometry & Vision Sciences, Cardiff University, Redwood Building, King Edward VII Avenue, Cardiff, CF10 3NB, Wales, UK

    Robert D. Young & Andrew J. Quantock

  2. Burnham Institute for Medical Research, North Torrey Pines Rd, La Jolla, CA, 92037, USA

    Tomoya O. Akama & Michiko N. Fukuda

  3. Moorfields Eye Hospital, 162 City Road, London, EC1V 2PD, UK

    Bruce Allan

  4. Division of Molecular Genetics, Institute of Ophthalmology, UCL, 11-43 Bath Street, London, EC1V 9EL, UK

    Petra Liskova & Neil D. Ebenezer

  5. Connective Tissue Biology Laboratories, School of Biosciences, Cardiff University, Museum Ave, Cardiff, CF10 3US, Wales, UK

    Briedgeen Kerr & Bruce Caterson

Authors
  1. Robert D. Young
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tomoya O. Akama
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Petra Liskova
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Neil D. Ebenezer
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bruce Allan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Briedgeen Kerr
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Bruce Caterson
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Michiko N. Fukuda
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Andrew J. Quantock
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Andrew J. Quantock.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Young, R.D., Akama, T.O., Liskova, P. et al. Differential immunogold localisation of sulphated and unsulphated keratan sulphate proteoglycans in normal and macular dystrophy cornea using sulphation motif-specific antibodies. Histochem Cell Biol 127, 115–120 (2007). https://doi.org/10.1007/s00418-006-0228-8

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00418-006-0228-8

Keywords

Search

Navigation