Atypical composition and ultrastructure of proteoglycans in the mouse corneal stroma

Young, Robert David ORCID: https://orcid.org/0000-0002-8300-8002, Tudor, D., Hayes, Anthony Joseph, Kerr, Briedgeen, Hayashida, Y., Nishida, K., Meek, Keith Michael Andrew ORCID: https://orcid.org/0000-0002-9948-7538, Caterson, Bruce ORCID: https://orcid.org/0000-0001-6016-0661 and Quantock, Andrew James ORCID: https://orcid.org/0000-0002-2484-3120 2005. Atypical composition and ultrastructure of proteoglycans in the mouse corneal stroma. Investigative Ophthalmology and Visual Science 46 (6) , pp. 1973-1978. 10.1167/iovs.04-1309

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Abstract

PURPOSE: Recently, gene-targeted strains of mice with null mutations for specific proteoglycans (PGs) have been used for investigations of the functional role of these molecules. In the present study, the corneal stroma of the mouse was examined to provide some baseline PG morphologies in this species. METHODS: Monoclonal antibodies to specific glycosaminoglycan (GAG) chain sulfation patterns were used to characterize PG composition in corneal extracts by SDS-PAGE and Western blot analysis and to identify their tissue distribution by immunofluorescence microscopy. PGs were also visualized by transmission electron microscopy after contrast enhancement with cationic dye fixation. RESULTS: Western blot analysis of pooled corneal extracts and immunofluorescence of tissue sections identified 4-sulfated, but not 6-sulfated, chondroitin sulfate/dermatan sulfate (CS/DS). Keratan sulfate (KS) was present only as a low-sulfated moiety. Electron microscopic histochemistry disclosed a complex array of corneal PGs present as (1) fine filaments radiating from collagen fibrils, and (2) elongate, straplike structures, running either along the fibril axis or weaving across the primary fibril orientation. These large structures were digested by chondroitinase ABC, but not by keratanase. CONCLUSIONS: KS in the mouse is predominantly undersulfated and generates an immunostaining pattern that differs from that observed in corneas of other mammalian species thus far investigated. The mouse cornea resembles other mammalian corneas in the presence of filamentous arrays of small, collagen-associated stromal PGs visualized by cationic dye staining. However, large dye-positive structures with a CS/DS component are also present and appear to be unique to the cornea of this species.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences Optometry and Vision Sciences
Subjects:	R Medicine > RE Ophthalmology
Additional Information:	Confirmation received by publisher on 21 February 2014 that publisher's pdf can be self-archived 6 months after publication.
Publisher:	Association for Research in Vision and Ophthalmology
ISSN:	0146-0404
Date of First Compliant Deposit:	30 March 2016
Last Modified:	18 May 2023 08:14
URI:	https://orca.cardiff.ac.uk/id/eprint/33446

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