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Journal of Histochemistry and Cytochemistry, Vol. 49, 1341-1350, November 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

Increased Expression of Tenascin-C-binding Epithelial Integrins in Human Bullous Keratopathy Corneas

Alexander V. Ljubimova, Mehrnoosh Saghizadeha, Robert Pytelab, Dean Sheppardb, and M. Cristina Kenneya
a Ophthalmology Research Laboratories, Burns & Allen Research Institute, Cedars–Sinai Medical Center, University of California Los Angeles School of Medicine Affiliate, Los Angeles, California
b Department of Medicine and Lung Biology Center/San Francisco General Hospital, University of California San Francisco, San Francisco, California

Correspondence to: Alexander V. Ljubimov, Ophthalmology Research Laboratories, Cedars–Sinai Medical Center, 8700 Beverly Boulevard, Davis-5069, Los Angeles, CA 90048. E-mail: ljubimov@cshs.org

We previously found an abnormal deposition of an extracellular matrix glycoprotein, tenascin-C (TN-C), in human corneas with pseudophakic/aphakic bullous keratopathy (PBK/ABK). In this work, we studied cellular TN-C receptors in normal and PBK/ABK corneas. Cryostat sections of normal and PBK/ABK corneas were stained by immuno-fluorescence for TN-C receptors: {alpha}2, {alpha}8, {alpha}9, {alpha}Vß3, ß1, and ß6 integrins, and annexin II. ß6 integrin mRNA levels were assessed by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) using ß2-microglobulin gene to normalize the samples. In PBK/ABK compared to normal corneas, relatively minor changes were observed for {alpha}2 and ß1 integrins, and for annexin II. {alpha}8, {alpha}9, and ß6 subunits of TN-C receptors, {alpha}8ß1 {alpha}9ß1, and {alpha}Vß6, respectively, were absent from normal central corneas but were found in the central epithelium of PBK/ABK corneas. ß6 integrin showed the most significant accumulation. It correlated best with the expression of TN-C rather than with the expression of other {alpha}Vß6 ligands, fibronectin, and vitronectin. RT-PCR analysis also showed elevated levels of ß6 mRNA in PBK/ABK compared to normal corneas. Therefore, accumulation of TN-C in PBK/ABK corneas was accompanied by an increased expression of its three binding integrins, especially {alpha}Vß6 in the corneal epithelium. The interaction of tenascin-C with these integrins may contribute to the fibrotic process that occurs in PBK/ABK corneas.

(J Histochem Cytochem 49:1341–1350, 2001)

Key Words: bullous keratopathy, corneal epithelium, tenascin-C, {alpha}Vß6 integrin, {alpha}8 integrin, {alpha}9 integrin, annexin II, RT-PCR, immunofluorescence


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