December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Membrane Associated Matrix Metalloproteinase (mt1-mmp) and Mmp-2 Gene Expression Are Selectively Induced by Hypoxia in Monkey Choroid Retinal Cells
Author Affiliations & Notes
  • GN Lambrou
    Clinique Ophtalmologique Univ Strasbourg France
  • P Ottino
    Neuroscience Center and Department of Ophthalmology Unoversity Health Science Center New Orleans LA
  • A Ottlecz
    Ophtha Research Novartis Ophthalmics AG Basle Switzerland
  • J Finley
    Neuroscience Center
    University Health Science Center New Orleans LA
  • NG Bazan
    Neuroscience Center and Department of Ophthalmology
    University Health Science Center New Orleans LA
  • HE P Bazan
    Neuroscience Center and Department of Ophthalmology
    University Health Science Center New Orleans LA
  • Footnotes
    Commercial Relationships    G.N. Lambrou, Novartis Ophthalmics AG F, E; P. Ottino, None; A. Ottlecz, Novartis Ophthalmics AG F, E; J. Finley, None; N.G. Bazan, Novartis Ophthalmics AG F, R; H.E.P. Bazan, Novartis Ophthalmics AG F, R.
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 2752. doi:
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    • Get Citation

      GN Lambrou, P Ottino, A Ottlecz, J Finley, NG Bazan, HE P Bazan; Membrane Associated Matrix Metalloproteinase (mt1-mmp) and Mmp-2 Gene Expression Are Selectively Induced by Hypoxia in Monkey Choroid Retinal Cells . Invest. Ophthalmol. Vis. Sci. 2002;43(13):2752.

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

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Abstract

Abstract: : Purpose: The extracellular matrix undergoes extensive changes during retinal pathoangiogenesis. MMPs may play an important role in ischemic retinal diseases in which neovascularization, migration and proliferation of vascular endothelial cells represent critical events in the progression of the disease. Here we investigated the expression of MMPs in monkey choroid retinal endothelial cells (RF/6A) exposed to hypoxia. Method: RF/6A cells (8 x 106, CRL-1780 from ATCC) were seeded onto Petri dishes (60 mm diameter) coated with collagen type-I and were grown in F12-K medium containing 5% fetal calf serum (FCS). When cells reached ∼90% confluency, the medium was replaced with hypoxic (degassed) medium containing 1% FCS and cells were placed in a cycling chamber and perfused with 95% N2 and 5% O2 at 37 0C for 2 hr. In post-hypoxia recovery experiments, cells were maintained in non-hypoxic media for an additional 2 hr. mRNA was extracted from endothelial cells using a SV total RNA isolation kit from Promega. Levels of gene expression for MMP-1, MT1-MMP, MMP-2, MMP-9 and the upstream stimulator of MMPs, urokinase plasminogen activator (uPA) were determined by real time PCR. All quantitations were normalized to the 18s rRNA endogenous control and changes in gene expression were assessed as relative to untreated controls. Results: Hypoxia triggered a 3-fold increase in the expression of the MT-MMP-1 and MMP-2 genes in monkey endothelial cells. Hypoxia produced no detectable changes in the expression of uPA, MMP-1 and MMP-9 mRNA in endothelial cells. In the post-hypoxia recovery period the induction of MT1-MMP and MMP-2 was decreased, whereas no changes were observed in the other MMPs and uPA. Conclusion: Hypoxia selectively induces MT1-MMP and MMP-2 transcription. Because MT1-MMP activates latent MMP-2, this increase in gene expression highlights a mechanism by which these proteases participate in the degradation of components of the extracellular matrix and promote retinal neovascularization.

Keywords: 483 neovascularization • 554 retina • 399 enzymes/enzyme inhibitors 
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