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AP Cullen, V Bantseev, JR Trevithick, JG Sivak; Correlation of Bovine Lens Optical Function and Metabolic Properties in Vitro . Invest. Ophthalmol. Vis. Sci. 2002;43(13):461.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Recent work revealed that rat lenses treated for 30 min with 65 µM carbonyl cyanide m-chlorophenylhydrazone (the mitochondrial depolarising agent, CCCP) developed opacities, compared to control lenses (Bantseev et al., 1999). This study tests the hypothesis that mitochondrial electron transport chain potential (Δ;ψm) in epithelial and superficial cortical lens fibre cells can be correlated with the lens optical function. Methods: Bovine lenses were exposed to 65 and 32.5 µM CCCP for 30 minutes, washed with saline and incubated in culture medium at 370C and 4-5% CO2. Lens optical function was analysed using the ScantoxTM In Vitro Lens Assay System (Harvard Apparatus, Holliston, MA) before exposure, immediately, 4, 8 and 24 hours after the exposure. After 24 hours the Δ;ψm of epithelial and superficial cortical fibre cells of the same lenses were analysed using 20 µM rhodamine 123 fluorescence and confocal microscope. Results: Lenses treated with 32.5 (n=14) and 65 (n=14) µM CCCP developed visible opacities, compared to controls (n=11). Progressive loss of sharp focus (as measured by the variability of back vertex distance) was most evident at the 8 hour scan point, increasing from 0.310.03 mm (SEM) initially, to 0.740.15 mm (SEM) for 32.5 µM (p<0.05) and from 0.260.03 mm (SEM) initially, to 1.610.31 mm (SEM) for 65 µM (p<0.05). Sharpness of focus of control lenses did not change significantly (0.340.04 mm (SEM) vs. 0.350.02 mm (SEM) at 8 hours). At 24 hours lens optical function returned to the level of controls. Confocal analysis after 24 hours showed a CCCP concentration-dependent decrease of the Δ;ψm of epithelial and superficial cortical fibre cells. Conclusions: The results of this study show that the Δ;ψm of epithelial and superficial cortical fibre cells is correlated with lens optical function. Moreover, both optical damage and the collapse of mitochondrial electron transport chain potential could recover to that of controls 24 hours post-exposure to 65 and 32.5 µM of CCCP. The results further suggest that mitochondrial integrity of the lens epithelium and superficial cortical fibres are closely correlated with lens optical function. Moreover, recovery of lens metabolic properties maybe necessary for the recovery of lens optical function.
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