May 2005
Volume 46, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2005
Integrin Expression in Laser–Induced CNV in Mice: A Microarray Study
Author Affiliations & Notes
  • M.A. Gamulescu
    Ophthalmology, University Clinic Regensburg, Regensburg, Germany
  • T. Triche
    Pathology and Laboratory Medicine, CHLA, USC, Los Angeles, CA
  • J. Zhou
    Pathology, USC, Los Angeles, CA
  • J. Buckley
    Preventive Medicine, Norris Cancer Center, USC, Los Angeles, CA
  • S.J. Ryan
    Ophthalmology, Doheny Eye Institute, USC, Los Angeles, CA
  • D.R. Hinton
    Pathology, USC, Los Angeles, CA
    Ophthalmology, Doheny Eye Institute, USC, Los Angeles, CA
  • Footnotes
    Commercial Relationships  M.A. Gamulescu, None; T. Triche, None; J. Zhou, None; J. Buckley, None; S.J. Ryan, None; D.R. Hinton, None.
  • Footnotes
    Support  BMBF–LPD 9901/8–52, NIH EYO1545, Arnold and Mabel Beckman Foundation
Investigative Ophthalmology & Visual Science May 2005, Vol.46, 3022. doi:
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      M.A. Gamulescu, T. Triche, J. Zhou, J. Buckley, S.J. Ryan, D.R. Hinton; Integrin Expression in Laser–Induced CNV in Mice: A Microarray Study . Invest. Ophthalmol. Vis. Sci. 2005;46(13):3022.

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

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Abstract

Abstract: : Purpose: Apart from dysregulation of angiogenic and anti–angiogenic factors in the retina and choriocapillaris, a disturbed interaction of retinal pigment epithelial (RPE) cells with their extracellular matrix is hypothized to play a role in the pathogenesis of choroidal neovascularization (CNV) in age–related macular degeneration (AMD). Integrins alphaV, alpha5, beta1, beta3 and beta5 were shown to play a role in cell adhesion and neo–angiogenesis. The purpose of this investigation was to determine the changes in gene expression of integrins and cell adhesion proteins related to angiogenesis during development and involution of laser–induced choroidal neovascularisaion (CNV) in the mouse model. Methods: CNV was induced in each eye of 30 mice (C57/BL6 strain) using 4 diode laser burns to the posterior pole. The mice were sacrificed at different time points ranging from 6 hours to 14 days after laser application. The retinal pigment epithelium (RPE) and choroid immediately around the laser spots, and in later stages including the CNV itself, were dissected. Total RNA was extracted, reverse transcribed to cDNA and amplified using MessageAmpTM (Ambion). Each sample was hybridized to a triplicate of gene arrays (Affymetrix, mouse 430A), and "treated" samples were compared to samples from untreated RPE/choroid. The hybridization signals were globally normalized and filtered and data was analysed using the Genetrix software. Real time PCR was used to confirm differential expression of selected genes. Results: Integrin alpha–chains (namely alphaV and alpha5) showed a strong up–regulation especially in the first hours post–laser, whereas beta–chains (namely beta5) showed a stronger upregulation 3–14 days after laser photocoagulation. Membrane–metalloproteinases (MMPs) were regulated over all time–points of the experiment, MMP 3 and 9 being the most prominent upregulated genes. Procollagen subtypes were upregulated beginning on day 1 after laser and peaking on day 7. Conclusions:Gene array analysis is a strong tool for global analysis of genes involved in the course of CNV formation and involution. In this model of laser–induced CNV–formation, strong integrin and MMP regulation could be shown during the whole course of the experiment. Blocking specific integrins and/or MMPs could be, beside the anti–angiogenetic approach, a further means to control CNV formation in AMD.

Keywords: gene microarray • age-related macular degeneration • gene/expression 
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