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Originally published In Press as doi:10.1167/iovs.07-1529 on March 31, 2008
 (Investigative Ophthalmology and Visual Science. 2008;49:2956-2963.)
© 2008 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.07-1529
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Decreased Expression of Peroxiredoxins in Fuchs’ Endothelial Dystrophy

Ula V. Jurkunas,1,2,3 Ian Rawe,1 Maya S. Bitar,1,3 Cheng Zhu,1,3 Deshea L. Harris,1,3 Kathryn Colby,1,2,3 and Nancy C. Joyce1,3

1From the Schepens Eye Research Institute, Boston, Massachusetts; the 2Massachusetts Eye and Ear Infirmary, Boston, Massachusetts; and the 3Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts.

PURPOSE. To compare the relative expression of peroxiredoxin (Prx) proteins in normal human corneal endothelium with endothelium in corneas affected by Fuchs’ endothelial dystrophy (FED) and between normal human endothelium and epithelial/stromal tissue.

METHODS. Human corneal endothelial cell-Descemet’s membrane (HCEC-DM) complexes from normal and FED corneal buttons were dissected from the epithelium/stroma. For proteomic analysis, HCEC-DM protein extracts were separated by using two-dimensional gel electrophoresis. Relative differences in protein spot density was analyzed. Proteins of interest, including Prx isoforms, were identified by MALDI-TOF (matrix-assisted desorption ionization-time of flight) mass spectrometry. Western blot analysis compared the relative expression of Prx isoforms in normal and FED endothelium and between normal endothelium and normal epithelium/stroma. Expression of Prx-2 mRNA was compared by using real-time PCR.

RESULTS. Proteomic analysis identified differences in the relative expression of Prx isoforms between normal and FED endothelium. Western blot analysis confirmed that expression of Prx-2, –3, and –5 was significantly decreased (P < 0.05) in FED cells. Normal HCECs expressed significantly (P < 0.05) higher levels of Prx-2 and –3 than did the epithelium/stroma. Expression of Prx-5 was not significantly different (P > 0.05) in the endothelium versus the epithelium/stroma. Real-time PCR analysis revealed that Prx-2 mRNA was significantly decreased (P = 0.027) in FED samples.

CONCLUSIONS. Prx proteins were identified in human corneal endothelium. The fact that Prx-2 and –3 were expressed at significantly higher levels in HCEC-DM compared with the epithelium/stroma reflects the different physiologic activities of individual corneal cell types. Significantly decreased expression of Prx-2, –3, and –5 in FED may suggest an alteration in the ability of endothelial cells to withstand oxidant-induced damage and may be closely related to the pathogenesis of this disease.






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