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First published online 8 April 2003
doi: 10.1242/jcs.00441

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Journal of Cell Science 116, 2157-2167 (2003)
doi: 10.1242/jcs.00441

Research Article

Requirement for Pax6 in corneal morphogenesis: a role in adhesion

Janine Davis1, Melinda K. Duncan2, W. Gerald Robison, Jr3 and Joram Piatigorsky1,*

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 {gamma}-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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