April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Ultrastructural Changes In The Outer Retina And Retinal Pigment Epithelium Of The ApoB100,LDLR-/- Mouse, A Murine Model Of Aging Of The Human Retina
Author Affiliations & Notes
  • Lionel Bretillon
    INRA, University of Burgundy, Eye & Nutrition Research Group, Dijon, France
  • Emilie Simon
    Eye & Nutrition Research Group, INRA, University of Burgundy, Dijon, France
  • Bernadette Kantelip
    Anatomopathology, Hospital, Besançon, France
  • Joël Michel
    INRA, University of Burgundy, Dimacell - Cell Imaging Platform, Dijon, France
  • Niyazi Acar
    INRA, University of Burgundy, Eye & Nutrition Research Group, Dijon, France
  • Alain M. Bron
    Ophthalmology, University Hospital, Dijon, France
  • Jeannine Lherminier
    INRA, University of Burgundy, Dimacell - Cell Imaging Platform, Dijon, France
  • Catherine P. Creuzot-Garcher
    Ophthalmology, University Hospital, Dijon, France
  • Footnotes
    Commercial Relationships  Lionel Bretillon, None; Emilie Simon, None; Bernadette Kantelip, None; Joël Michel, None; Niyazi Acar, None; Alain M. Bron, None; Jeannine Lherminier, None; Catherine P. Creuzot-Garcher, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 957. doi:
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      Lionel Bretillon, Emilie Simon, Bernadette Kantelip, Joël Michel, Niyazi Acar, Alain M. Bron, Jeannine Lherminier, Catherine P. Creuzot-Garcher; Ultrastructural Changes In The Outer Retina And Retinal Pigment Epithelium Of The ApoB100,LDLR-/- Mouse, A Murine Model Of Aging Of The Human Retina. Invest. Ophthalmol. Vis. Sci. 2011;52(14):957.

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Abstract
 
Purpose:
 

Age-related Macular Degeneration (AMD) is the leading cause of visual impairment after the age of 50 years in Western countries. Deposition of lipids including cholesteryl esters within Bruch’s membrane (BrM) is a hallmark of aging of the human retina and is observed in eyes of patients with maculopathies and AMD. We previously demonstrated that the ApoB100,LDLR-/- mouse exhibits deposits of cholesteryl esters in BrM, fundus autofluorescence and reduced electroretinographic response, making it a relevant model of aging of the human retina. In order to better characterize its phenotype, we aimed to evaluate the ultrastructural changes in the retina of the ApoB100,LDLR-/- mouse.

 
Methods:
 

In vivo fundus autofluorescence of aged ApoB100,LDLR-/- mice was evaluated using a confocal scanning laser ophthalmoscope. Mices were enucleated and the ocular globes were immediately fixed with 2.5% glutaraldehyde prepared in 0.1M phosphate buffer pH7.2 for 20h, postfixed with 0.5% buffered osmium tetroxide for 1h, dehydrated through a graded ethanol series and propylene oxide, and embedded in Epon resin. Semithin sections (0.5µm) were stained with 0.2% toluidine blue pH11 prior to examination by bright field microscopy with a Leica DRMB microscope. Ultrathin sections (90nm) were counterstained with 3% (w/v) uranyl acetate and lead citrate and examined with a Hitachi H7500 transmission electron microscope operating at 80kV and equipped with an AMT camera.

 
Results:
 

ApoB100,LDLR-/- mice showed punctuated fundus autofluorescence at 488nm, consistent with the accumulation of lipid-rich lipofuscin material. ApoB100,LDLR-/- mice exhibited irregular disk arrangement of the outer segments of photoreceptors. Accumulation of remnants of incomplete phagocytosis of photoreceptor outer segments was visible in the RPE. Vacuoles were observed in the cytoplasm and mitochondria of the RPE. Electron-dense material accumulated in the RPE. Enlarged basal infoldings were found in ApoB100,LDLR-/- mice, and material with amorphous deposits can be seen between basal infoldings in the mouse RPE.

 
Conclusions:
 

Our data on ultrastructural changes in the outer retina and RPE of aged ApoB100,LDLR-/- mice support further investigations on the use of the ApoB100,LDLR-/- mouse as a relevant model for dry AMD.

 
Keywords: aging • microscopy: electron microscopy • age-related macular degeneration 
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