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Om P Srivastava, Kiran Srivastava, Shylaja Hegde, Roy Joseph, ; Temporal Changes in Lenses of CRYAAN101D Transgenic Mice Compared to CRYAAWT Mice Prior to Cataract Development. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1207.
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© ARVO (1962-2015); The Authors (2016-present)
The purpose was to examine temporal changes in lenses of CRYAAN101D mice relative to CRYAAWT (wild type, WT) mice to elucidate potential mechanism of cataract development.
Because the lenses of mice expressing αA N101D transgene developed mild cortical opacity by the age of 7-months (J. Biol. Chem. 286:11579-11592, 2011), the relative temporal changes in lenses 1-, 3- and 5-months old transgenic mice compared to WT mice were determined. The analyses included comparative changes in fiber cells ultrastructure including cytoskeleton and membranes, and binding of αAN101D to membranes and its distribution in water soluble (WS)- and water insoluble (WI)-proteins.
The scanning electron microscopic (SEM) analysis showed altered fiber cell structures in 5-months old lenses of CRYAAN101D mice. Immunohistochemical analyses using anti-aquaporin 0-, anti-His (for αA identification)- and anti-phalloidin-antibodies revealed that membranes and cytoskeleton of lenses αAN101D mice were altered relative to age-matched WT lenses. Similar comparative analysis showed a greater level of association of αAN101D to lens membranes of transgenic mice relative to WT mice, which was further confirmed by EM-immunogold labeling analysis of lens sections. The comparative protein profiles revealed greater levels of water insolubilization and degradation of αAN101D in lenses of αAN101D mice.
Relative to WT-mice, the lenses of αAN101D mice showed altered membranes and cytoskeleton, an increased association of αAN101D to membranes and water insolubilzation and degradation. These changes might play a role in the development of lens opacity in CRYAAN101D mice.
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