May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Identification of Lipid Oxidation Products and Proteins in Bruch’s Membrane from Normal and AMD Donor Eyes
X. Gu; K. Shadrach; M. Sun; K.A. West; L. Shan; S. Hazan; R.G. Salomon; J.G. Hollyfield; J.W. Crabb
Author Affiliations & Notes
  • X. Gu
    Ophthalmic Research, Cleveland Clinic Found, Cleveland, OH, United States
  • K. Shadrach
    Ophthalmic Research, Cleveland Clinic Found, Cleveland, OH, United States
  • M. Sun
    Chemistry, Case Western Reserve University, Cleveland, OH, United States
  • K.A. West
    Chemistry, Case Western Reserve University, Cleveland, OH, United States
  • L. Shan
    Cell Biology, Cleveland Clinic Found, Cleveland, OH, United States
  • S. Hazan
    Cell Biology, Cleveland Clinic Found, Cleveland, OH, United States
  • R.G. Salomon
    Cell Biology, Cleveland Clinic Found, Cleveland, OH, United States
  • J.G. Hollyfield
    Cell Biology, Cleveland Clinic Found, Cleveland, OH, United States
  • J.W. Crabb
    Cell Biology, Cleveland Clinic Found, Cleveland, OH, United States
  • Footnotes
    Commercial Relationships  X. Gu, None; K. Shadrach, None; M. Sun, None; K.A. West, None; L. Shan, None; S. Hazan, None; R.G. Salomon, None; J.G. Hollyfield, None; J.W. Crabb, None.
  • Footnotes
    Support  EY06603, EY14239, GM21249, EY02362, EY14240, Funds from Foundation Fighting Blindness and CCF
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 1622. doi:
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      X. Gu, K. Shadrach, M. Sun, K.A. West, L. Shan, S. Hazan, R.G. Salomon, J.G. Hollyfield, J.W. Crabb; Identification of Lipid Oxidation Products and Proteins in Bruch’s Membrane from Normal and AMD Donor Eyes . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1622.

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Abstract

Abstract: : Purpose: To identify lipid oxidation products and proteins in Bruch’s membrane possibly associated with the pathogenesis of age-related macular degeneration (AMD). Methods: Bruch’s membrane was isolated free of adjacent tissues and in the presence of antioxidants from 5 normal and 4 AMD donor eyes. Lipids were extracted with chloroform and methanol and analyzed by LC MS. Proteins were subjected to SDS-PAGE, blotted to PVDF membranes and/or gel bands were excised and proteins identified by LC MS/MS. Western analyses were used to screen for oxidative protein modifications. Results: Lipid oxidation products from docosahexaenoyl phosphatidylcholine (DHA-PC), arachidonoyl (AA)-PC, and linoleyl (LA)-PC were detected in relatively greater amounts from AMD than age matched healthy donor eyes. Common drusen proteins were also detected in Bruch’s membrane, including tissue inhibitor metalloproteinase-3, clusterin, vitronectin and serum albumin. Carboxyethylpyrrole protein adducts generated from the oxidation of DHA containing lipids were more abundant in Bruch’s membrane from AMD donors. Conclusions: These observations are consistent with recent drusen analyses (2002 Proc. Natl. Acad. Sci. USA 99: 14682) and support a role for lipid oxidation products in Bruch’s membrane changes associated with AMD.

Keywords: Bruch's membrane • age-related macular degeneration • oxidation/oxidative or free radical damage 
© 2003, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2015 Association for Research in Vision and Ophthalmology.
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