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Glycobiology, 2000, Vol. 10, No. 3 305-312
© 2000 Oxford University Press

Ultrastructural localization of sulfated and unsulfated keratan sulfate in normal and macular corneal dystrophy type I

David Lewis, Yvonne Davies, Ian A. Nieduszynski, Fiona Lawrence, Andrew J. Quantock2, Richard Bonshek3 and Nigel J. Fullwood1

Department of Biological Sciences, Institute of Environmental and Natural Sciences, Lancaster University, Lancaster LA1 4YQ, UK, 2Department of Optometry and Vision Sciences, Cardiff University, Cardiff, UK, and 3University of Manchester and Royal Manchester Eye Hospital, Manchester, UK

Keratan sulfate (KS) proteoglycans are of importance for the maintenance of corneal transparency as evidenced in the condition macular corneal dystrophy type I (MCD I), a disorder involving the absence of KS sulfation, in which the cornea becomes opaque. In this transmission electron microscope study quantitative immuno- and histochemical methods have been used to examine a normal and MCD I cornea. The monoclonal antibody, 5-D-4, has been used to localize sulfated KS and the lectin Erythrina cristagalli agglutinin (ECA) to localize poly N-acetyllactosamine (unsulfated KS). In normal cornea high levels of sulfated KS were detected in the stroma, Bowman’s layer, and Descemet’s membrane and low levels in the keratocytes, epithelium and endothelium. Furthermore, in normal cornea, negligible levels of labeling were found for N-acetyllactosamine (unsulfated KS). In the MCD I cornea sulfated KS was not detected anywhere, but a specific distribution of N-acetyllactosamine (unsulfated KS) was evident: deposits found in the stroma, keratocytes, and endothelium labeled heavily as did the disrupted posterior region of Descemet’s membrane. However, the actual cytoplasm of cells and the undisrupted regions of stroma revealed low levels of labeling. In conclusion, little or no unsulfated KS is present in normal cornea, but in MCD I cornea the abnormal unsulfated KS was localized in deposits and did not associate with the collagen fibrils of the corneal stroma. This study has also shown that ECA is an effective probe for unsulfated KS.

1 To whom correspondence should be addressed


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