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First published online 8 April 2003
doi: 10.1242/jcs.00441
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Research Article |
1 Laboratory of Molecular and Developmental Biology, National Eye Institute,
National Institutes of Health, Bethesda, MD 20892-2730, USA
2 Department of Biological Sciences, University of Delaware, Newark, DE 19716,
USA
3 Laboratory of Mechanisms of Ocular Disease, National Eye Institute, National
Institutes of Health, Bethesda, MD 20892-2735, USA
* Author for correspondence (e-mail: joramp{at}nei.nih.gov)
Accepted 17 February 2003
The Pax6 transcription factor functions early during embryogenesis to
control key steps in brain, pancreas, olfactory and ocular system development.
A requirement for Pax6 in proper formation of lens, iris and retina is well
documented. By examining the corneas of heterozygous Small eye (SEY) mice,
this report shows that Pax6 is also necessary for normal corneal
morphogenesis. In particular, the epithelial component of the postnatal and
adult SEY (+/–) cornea is thinner owing to a reduction in the number of
cell layers, despite a tenfold increase in the proliferative index and no
change in TUNEL labeling. Ultrastructural views revealed large gaps between
corneal epithelial cells and a change in the appearance of desmosomes,
suggesting that adhesion abnormalities contribute to the corneal phenotype of
SEY (+/–) mice. Western blot analysis and immunofluorescence showed
equivalent amounts and normal localization of E-cadherin in SEY (+/–)
corneas, and the actin cytoskeleton appeared normal as judged by phalloidin
staining. By contrast, the levels of desmoglein, ß-catenin and
-catenin were reduced in the SEY (+/–) cornea. In addition, the
amount of keratin-12 mRNA and protein, the major intermediate filament, was
reduced in SEY (+/–) corneal epithelium as shown by in situ
hybridization and immunohistochemistry. Finally, the SEY (+/–) corneal
epithelium adheres less well than wild-type when challenged with gentle
rubbing using a microsponge. In conclusion, our results indicate that cellular
adhesion is compromised in the SEY (+/–) corneal epithelium and suggests
a role for Pax6 in the proper generation and maintenance of the adult
cornea.
Key words: Pax6, Cornea, Adhesion, SEY mice, Development
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