April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Proteomic analysis of sub-retinal deposits and RPE of the ApoB100,LDLR-/- mouse, a murine model of aging of the human retina
Author Affiliations & Notes
  • Lionel Bretillon
    Centre des Sciences du Goût et de l'Alimentation, UMR1324 INRA, 6265 CNRS, Université de Bourgogne, Eye & Nutrition Research Group, Dijon, France
  • Emilie Simon
    Centre des Sciences du Goût et de l'Alimentation, UMR1324 INRA, 6265 CNRS, Université de Bourgogne, Eye & Nutrition Research Group, Dijon, France
  • Christine Arnould
    DImaCell Plate-forme Imagerie Cellulaire, INRA, Université de Bourgogne, Dijon, France, Dijon, France
  • Géraldine Lucchi
    Plate-forme protéomique CLIPP, CHU, Université de Bourgogne, Dijon, France
  • Delphine Pecqueur
    Plate-forme protéomique CLIPP, CHU, Université de Bourgogne, Dijon, France
  • Caroline Truntzer
    Plate-forme protéomique CLIPP, CHU, Université de Bourgogne, Dijon, France
  • Niyazi Acar
    Centre des Sciences du Goût et de l'Alimentation, UMR1324 INRA, 6265 CNRS, Université de Bourgogne, Eye & Nutrition Research Group, Dijon, France
  • Jeannine Lherminier
    DImaCell Plate-forme Imagerie Cellulaire, INRA, Université de Bourgogne, Dijon, France, Dijon, France
  • Patrick Ducoroy
    Plate-forme protéomique CLIPP, CHU, Université de Bourgogne, Dijon, France
  • Catherine P Garcher
    Centre des Sciences du Goût et de l'Alimentation, UMR1324 INRA, 6265 CNRS, Université de Bourgogne, Eye & Nutrition Research Group, Dijon, France
    Department of Ophthalmology, University Hospital, Dijon, France
  • Footnotes
    Commercial Relationships Lionel Bretillon, None; Emilie Simon, None; Christine Arnould, None; Géraldine Lucchi, None; Delphine Pecqueur, None; Caroline Truntzer, None; Niyazi Acar, None; Jeannine Lherminier, None; Patrick Ducoroy, None; Catherine Garcher, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 1316. doi:
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      Lionel Bretillon, Emilie Simon, Christine Arnould, Géraldine Lucchi, Delphine Pecqueur, Caroline Truntzer, Niyazi Acar, Jeannine Lherminier, Patrick Ducoroy, Catherine P Garcher; Proteomic analysis of sub-retinal deposits and RPE of the ApoB100,LDLR-/- mouse, a murine model of aging of the human retina. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1316.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose
 

The deposition of basal deposits and drusen at the basement of the retinal pigment epithelium (RPE) and within Bruch’s Membrane (BrM) is a hallmark of aging and is enhanced in Age-related Macular Degeneration (AMD). The question of the origin of the deposits and drusen remains largely unresolved. Part of the lipid content of drusen and basal deposits originates from the choroid. Recently, sub-retinal drusenoid deposits (SDD) were characterized in AMD eyes. Data from a topographical study in human eyes suggest that rods may be the main source of SDD whereas basal deposits may originate from cones. To improve our knowledge on the pathophysiology of aging and mechanisms of lipid deposition, we characterized the proteomic changes in the retina and RPE of the ApoB100,LDLR-/- mouse, a relevant model of aging of the human retina.

 
Methods
 

Ocular globes of aged ApoB100,LDLR-/- and control mice were embedded in OCT and cryosectioned in 10µm-thick slices. The outer segments of the photoreceptors, SDD and RPE samples were collected by Laser Capture Microdissection. Proteins were extracted and purified by 1D gel electrophoresis. After trypsin digestion, peptides were analyzed by nanoLC-MS/MS using an LTQ-Orbitrap Elite mass spectrometer. The proteins were identified using a combination of X!Tandem and Mascot, and then validated using PeptideProphet and ProteinProphet. Data were processed and quantified using a set of home-made and open-source software tools. Differential proteins were selected using the MSstats library on R-software with a 5% FDR, and searched against Kegg and Reactome pathways and Gene Ontology terms databases using the Cytoscape software.

 
Results
 

From 38 to 96 proteins were identified in the different samples. Lower levels of retinol dehydrogenase and up-regulation of S-arrestin in ApoB100,LDLR-/- mice compared to controls may explain reduced electroretinographic response in the transgenic mouse (impaired retinoid cycle). Up-regulation of crystallins in the transgenic mouse (in outer segments, sub-retinal deposits and RPE) is consistent with proteomic data in eyes collected from patients with AMD showing increased levels of crystallins αA and αB.

 
Conclusions
 

The proteomic data are consistent with findings in AMD patients and may add possible clues to explain functional changes in the ApoB100,LDLR-/- mouse (effect on S-arrestin and retinol dehydrogenase).

 
Keywords: 413 aging • 663 proteomics  
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