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Quantitative Proteomic Analysis of Retina in Oxygen-Induced Retinopathy Mice using iTRAQ with 2D NanoLC-nanoESI-MS/MS

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Author(s): Lei Zhou | Liu Xinling | Siew Kwan Koh | Li Xiaorong | Roger W. Beuerman

Journal: Journal of Integrated OMICS
ISSN 2182-0287

Volume: 1;
Issue: 2;
Start page: 226;
Date: 2011;
Original page

Keywords: Retina neovascularization | Oxygen-induced retinopathy | Retina proteome | iTRAQ | 2D nano LC-nanoESI-MS/MS

ABSTRACT
Analysis of the retina proteome during hypoxia-induced retinal neovascularization (RN) process may be helpful to elucidate pathogenesis of related diseases, such as diabetic retinopathy (DR) and retinopathy of prematurity (ROP). Retinal neovasculariation was induced in 7-day old, C57BL/6 mice by exposure to 80% oxygen for 5 days followed by 5 days in room air. Retinas from mice at postnatal day 17 from control and oxygen-induced retinopathy (OIR) groups were used for proteomic analysis. We have employed a quantitative proteomic approach, iTRAQ (isobaric Tagging for Relative and Absolute protein Quantification) coupled with 2D nanoLC-nanoESI-MS/MS to quantitatively compare the relative changes in the retina proteome from control and OIR mice. In total, 264 protein groups were identified at a 95% confidence level. Among them, OIR induced significant changes in 28 proteins (14 up-regulated and 14 down-regulated). Obvious changes include the up-regulation of a few plasma proteins (i.e. serum albumin, hemoglobin), indicating the breakdown of the blood-retina barrier. Vimentin, ribosomal proteins, some proteases, neural cell adhesion molecule (NCAM) 180, retinoschisin were found to be up-regulated and several crystallins such as isoform 1 of α crystallin A chain, isoform 2 of α crystallin A chain, α crystallin B chain, γ crystallin D and β-A3/A1 crystallin were down-regulated. The iTRAQ result of α crystallin B chain was also verified by Western blot analysis. The proteomic results from this study provide new avenues for understanding the pathogenesis of OIR induced retinal neovascularization and related retina diseases.
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