June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Mice Expressing CRYAAN101D Show Altered Membrane Structure and Increased Membrane-Association of Deamidated αA Compared to CRYAA Wild-Type (WT) Mice
Author Affiliations & Notes
  • Kiran Srivastava
    Vision Sciences, University of Alabama at Birmingham, Birmingham, AL
  • Roy Joseph
    Vision Sciences, University of Alabama at Birmingham, Birmingham, AL
  • Shylaja Hegde
    Vision Sciences, University of Alabama at Birmingham, Birmingham, AL
  • Om Srivastava
    Vision Sciences, University of Alabama at Birmingham, Birmingham, AL
  • Footnotes
    Commercial Relationships Kiran Srivastava, None; Roy Joseph, None; Shylaja Hegde, None; Om Srivastava, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 5747. doi:
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      Kiran Srivastava, Roy Joseph, Shylaja Hegde, Om Srivastava, ; 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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      © ARVO (1962-2015); The Authors (2016-present)

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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  
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