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Hypoxia-Ischemia or Excitotoxin-Induced Tissue Plasminogen Activator- Dependent Gelatinase Activation in Mice Neonate Brain Microvessels

Hypoxia-ischemia (HI) and excitotoxicity are validated causes of neonatal brain injuries and tissue plasminogen activator (t-PA) participates in the processes through proteolytic and receptor-mediated pathways. Brain microvascular endothelial cells from neonates in culture, contain and release more...

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Autores principales: Omouendze, Priscilla L., Henry, Vincent J., Porte, Baptiste, Dupré, Nicolas, Carmeliet, Peter, Gonzalez, Bruno J., Marret, Stéphane, Leroux, Philippe
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3735506/
https://www.ncbi.nlm.nih.gov/pubmed/23940734
http://dx.doi.org/10.1371/journal.pone.0071263
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author Omouendze, Priscilla L.
Henry, Vincent J.
Porte, Baptiste
Dupré, Nicolas
Carmeliet, Peter
Gonzalez, Bruno J.
Marret, Stéphane
Leroux, Philippe
author_facet Omouendze, Priscilla L.
Henry, Vincent J.
Porte, Baptiste
Dupré, Nicolas
Carmeliet, Peter
Gonzalez, Bruno J.
Marret, Stéphane
Leroux, Philippe
author_sort Omouendze, Priscilla L.
collection PubMed
description Hypoxia-ischemia (HI) and excitotoxicity are validated causes of neonatal brain injuries and tissue plasminogen activator (t-PA) participates in the processes through proteolytic and receptor-mediated pathways. Brain microvascular endothelial cells from neonates in culture, contain and release more t-PA and gelatinases upon glutamate challenge than adult cells. We have studied t-PA to gelatinase (MMP-2 and MMP-9) activity links in HI and excitotoxicity lesion models in 5 day–old pups in wild type and in t-PA or its inhibitor (PAI-1) genes inactivated mice. Gelatinolytic activities were detected in SDS-PAGE zymograms and by in situ fluorescent DQ-gelatin microscopic zymographies. HI was achieved by unilateral carotid ligature followed by a 40 min hypoxia (8%O(2)). Excitotoxic lesions were produced by intra parenchymal cortical (i.c.) injections of 10 µg ibotenate (Ibo). Gel zymograms in WT cortex revealed progressive extinction of MMP-2 and MMP-9 activities near day 15 or day 8 respectively. MMP-2 expression was the same in all strains while MMP-9 activity was barely detectable in t-PA(−/−) and enhanced in PAI-1(−/−) mice. HI or Ibo produced activation of MMP-2 activities 6 hours post-insult, in cortices of WT mice but not in t-PA(−/−) mice. In PAI-1(−/−) mice, HI or vehicle i.c. injection increased MMP-2 and MMP-9 activities. In situ zymograms using DQ-gelatin revealed vessel associated gelatinolytic activity in lesioned areas in PAI-1(−/−) and in WT mice. In WT brain slices incubated ex vivo, glutamate (200 µM) induced DQ-gelatin activation in vessels. The effect was not detected in t-PA(−/−)mice, but was restored by concomitant exposure to recombinant t-PA (20 µg/mL). In summary, neonatal brain lesion paradigms and ex vivo excitotoxic glutamate evoked t-PA-dependent gelatinases activation in vessels. Both MMP-2 and MMP-9 activities appeared t-PA-dependent. The data suggest that vascular directed protease inhibition may have neuroprotection potential against neonatal brain injuries.
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spelling pubmed-37355062013-08-12 Hypoxia-Ischemia or Excitotoxin-Induced Tissue Plasminogen Activator- Dependent Gelatinase Activation in Mice Neonate Brain Microvessels Omouendze, Priscilla L. Henry, Vincent J. Porte, Baptiste Dupré, Nicolas Carmeliet, Peter Gonzalez, Bruno J. Marret, Stéphane Leroux, Philippe PLoS One Research Article Hypoxia-ischemia (HI) and excitotoxicity are validated causes of neonatal brain injuries and tissue plasminogen activator (t-PA) participates in the processes through proteolytic and receptor-mediated pathways. Brain microvascular endothelial cells from neonates in culture, contain and release more t-PA and gelatinases upon glutamate challenge than adult cells. We have studied t-PA to gelatinase (MMP-2 and MMP-9) activity links in HI and excitotoxicity lesion models in 5 day–old pups in wild type and in t-PA or its inhibitor (PAI-1) genes inactivated mice. Gelatinolytic activities were detected in SDS-PAGE zymograms and by in situ fluorescent DQ-gelatin microscopic zymographies. HI was achieved by unilateral carotid ligature followed by a 40 min hypoxia (8%O(2)). Excitotoxic lesions were produced by intra parenchymal cortical (i.c.) injections of 10 µg ibotenate (Ibo). Gel zymograms in WT cortex revealed progressive extinction of MMP-2 and MMP-9 activities near day 15 or day 8 respectively. MMP-2 expression was the same in all strains while MMP-9 activity was barely detectable in t-PA(−/−) and enhanced in PAI-1(−/−) mice. HI or Ibo produced activation of MMP-2 activities 6 hours post-insult, in cortices of WT mice but not in t-PA(−/−) mice. In PAI-1(−/−) mice, HI or vehicle i.c. injection increased MMP-2 and MMP-9 activities. In situ zymograms using DQ-gelatin revealed vessel associated gelatinolytic activity in lesioned areas in PAI-1(−/−) and in WT mice. In WT brain slices incubated ex vivo, glutamate (200 µM) induced DQ-gelatin activation in vessels. The effect was not detected in t-PA(−/−)mice, but was restored by concomitant exposure to recombinant t-PA (20 µg/mL). In summary, neonatal brain lesion paradigms and ex vivo excitotoxic glutamate evoked t-PA-dependent gelatinases activation in vessels. Both MMP-2 and MMP-9 activities appeared t-PA-dependent. The data suggest that vascular directed protease inhibition may have neuroprotection potential against neonatal brain injuries. Public Library of Science 2013-08-06 /pmc/articles/PMC3735506/ /pubmed/23940734 http://dx.doi.org/10.1371/journal.pone.0071263 Text en © 2013 Omouendze et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Omouendze, Priscilla L.
Henry, Vincent J.
Porte, Baptiste
Dupré, Nicolas
Carmeliet, Peter
Gonzalez, Bruno J.
Marret, Stéphane
Leroux, Philippe
Hypoxia-Ischemia or Excitotoxin-Induced Tissue Plasminogen Activator- Dependent Gelatinase Activation in Mice Neonate Brain Microvessels
title Hypoxia-Ischemia or Excitotoxin-Induced Tissue Plasminogen Activator- Dependent Gelatinase Activation in Mice Neonate Brain Microvessels
title_full Hypoxia-Ischemia or Excitotoxin-Induced Tissue Plasminogen Activator- Dependent Gelatinase Activation in Mice Neonate Brain Microvessels
title_fullStr Hypoxia-Ischemia or Excitotoxin-Induced Tissue Plasminogen Activator- Dependent Gelatinase Activation in Mice Neonate Brain Microvessels
title_full_unstemmed Hypoxia-Ischemia or Excitotoxin-Induced Tissue Plasminogen Activator- Dependent Gelatinase Activation in Mice Neonate Brain Microvessels
title_short Hypoxia-Ischemia or Excitotoxin-Induced Tissue Plasminogen Activator- Dependent Gelatinase Activation in Mice Neonate Brain Microvessels
title_sort hypoxia-ischemia or excitotoxin-induced tissue plasminogen activator- dependent gelatinase activation in mice neonate brain microvessels
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3735506/
https://www.ncbi.nlm.nih.gov/pubmed/23940734
http://dx.doi.org/10.1371/journal.pone.0071263
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