Differential immunogold localisation of sulphated and unsulphated keratan sulphate proteoglycans in normal and macular dystrophy cornea using sulphation motif-specific antibodies |
| |
Authors: | Robert D Young Tomoya O Akama Petra Liskova Neil D Ebenezer Bruce Allan Briedgeen Kerr Bruce Caterson Michiko N Fukuda Andrew J Quantock |
| |
Institution: | (1) Structural Biophysics Group, School of Optometry & Vision Sciences, Cardiff University, Redwood Building, King Edward VII Avenue, Cardiff, CF10 3NB, Wales, UK;(2) Burnham Institute for Medical Research, North Torrey Pines Rd, La Jolla, CA 92037, USA;(3) Moorfields Eye Hospital, 162 City Road, London, EC1V 2PD, UK;(4) Division of Molecular Genetics, Institute of Ophthalmology, UCL, 11-43 Bath Street, London, EC1V 9EL, UK;(5) Connective Tissue Biology Laboratories, School of Biosciences, Cardiff University, Museum Ave, Cardiff, CF10 3US, Wales, UK |
| |
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. |
| |
Keywords: | Cornea Keratan sulphate proteoglycan Macular corneal dystrophy Sulphation motif-specific antibodies Immunolocalisation |
本文献已被 PubMed SpringerLink 等数据库收录! |
|