Abstract
Purpose:
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.
Methods:
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.
Results:
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.
Conclusions:
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.
Keywords: 488 crystallins •
445 cataract •
554 immunohistochemistry