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Kiran Srivastava, Roy Joseph, Shylaja Hegde, Om Srivastava, Lens Catractogenesis; Mice Expressing CRYAAN101D Show Altered Membrane Structure and Increased Membrane-Association of Deamidated αA Compared to CRYAA Wild-Type (WT) Mice. Invest. Ophthalmol. Vis. Sci. 2013;54(15):5747.
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The purpose was to determine whether membrane structures and membrane-association of deamidated αA in lenses of αAN101D mice expressing αAN101D transgene were altered relative to wild type (WT) αA transgenic mice.
We have shown that mice expressing deamidated alpha A-crystallin (αAN101D) transgene developed cortical cataract by the age of 7-months compared to the age-matched mice expressing WT αA-transgene (J. Biol. Chem. 286:11579-11592, 2011). To examine membrane structures, the axial and equatorial sections of lenses from 3- and 5-month-old transgenic- and WT-mice were immunohistologically analyzed using anti-aquaporin 0- (to identify intact membrane) and anti-His- (to identify His-tagged αAN101D) antibodies. Additionally, isolated membranes from these lenses were examined for the relative association of αAN101D WT-αA by the Western blot method using anti-His- and anti-aquaporin 0-antibodies.
The immunohistochemical results showed that orientation of fiber cells, their sizes along with outer cortical fiber cell membrane structures, and membrane-association of αA N101D were altered in lenses of 3- and 5-month-old transgenic mice relative to the lenses of same aged WT mice. Further, the Western blot analysis showed that lens membranes of 5-month-old transgenic mice had a greater association with αA N101D relative to the membranes of lenses of wild-type mice.
The results suggest that a relatively greater association of αAN101D with lens membranes of outer coritical fibers in transgenic mice relative to WT-mice might lead to membrane disruption and cortical cataract.
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