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YS Oleynikov, F Sun, A Spector; Cell Morphology and Cytoskeletal Disruption in Lens Epithelial Cell Lines Subjected to Peroxide Stress . Invest. Ophthalmol. Vis. Sci. 2002;43(13):2373.
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Purpose: Oxidation due to elevation of peroxide concentration in the aqueous, vitreous and the lens is a well-known mechanism leading to cataract formation. This laboratory has developed and studied lens epithelial cell lines that have been conditioned to survive peroxide exposure. In a recent study, it was found that major changes in gene expression profile such as a marked downregulation of alpha-actin and other associated cytoskeletal proteins occur in these cells. In this study, we examine at high-resolution the changes in lens epithelial cell morphology and cytoskeleton in cell lines conditioned to survive either hydrogen peroxide or a lipid peroxide prototype, tertiary butyl hydroperoxide (TBHP). Methods: The normal immortal murine lens epithelial cell line αTN4-1 has been conditioned to survive 125uM hydrogen peroxide, 100uM TBHP or both peroxides. The conditioned cells were propagated in cell culture and fixed on glass coverslips. Cytoskeletal components were detected with specific antibodies and imaged using high-resolution fluorescence microscopy. Results: We find major disruption of the actin cytoskeleton affecting lens epithelial cell shape, cell spreading and cytoplasmic organization. The disruption of stress fibers results in accumulation of shorter actin filaments and their fragments throughout the cell. Unable to maintain viable stress fibers, the cells accumulate disorganized actin patches and conglomerate throughout the cytoplasm and especially at focal adhesions. Cells are poorly spread and are sometimes found in multicellular aggregates rather than in a monolayer. Microtubules are not disrupted or severely affected. The change is persistent and characteristic of these new cell lines, suggesting a major shift in phenotype and gene expression profile. Conclusion: Peroxide conditioning of lens epithelial cells results in major disruption of the actin cytoskeleton and, consequently, cell morphology. These changes do not appear to be related to resistance to peroxide stress. The results suggest that epithelial cells in the in vivo lens that survive peroxide stress may be recognized by modification of their intracellular morphology. If disorganized actin structures form in vivo in response to peroxide stress, they may serve as scatter points contributing to a loss of transparency.
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