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J.L. Robben, A. Jergensmeir, D. Lin, L. Takemoto, S. Lobell, D.J. Takemoto; Expression of SOD in Whole Lens Prevents Cataract Formation . Invest. Ophthalmol. Vis. Sci. 2005;46(13):4697.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To determine if overexpression of superoxide dismutase (SOD) in rat whole lens cultures will prevent oxidative damage, changes in gap junction activity and cataract formation. Methods: Whole rat lenses were incubated with 50µg wild type gfpSOD–1 plasmid for 24 hr prior to incubation for another 24 hr with 100µM hydrogen peroxide. Cataract formation was determined by grid analyses. Presence of SOD protein was determined by Western blot, gap junction activity was measured by dye transfer, and location of the SOD in the intact lens was determined by measurement of the gfp fluorescence using confocal microscopy. Results: SOD protein was expressed in whole rat lens in culture after 24 hrs as determined by Western blot using anti–gfp antisera. Fluorescent gfp was measurable at 100–200 µm into the bow region. Expression of the SOD protein correlated with prevention of hydrogen peroxide induced cataract formation. Inhibition of gap junction activity observed with hydrogen peroxide treatment was prevented by overexpression of SOD. Conclusions: SOD plasmid constructs can enter the whole lens, result in the overexpression of SOD protein, and this can protect whole lens from hydrogen peroxide–induced cataract formation. Plasmid delivery of protective SOD provides a method of cataract prevention in whole lens in culture.
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