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Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart

AIMS: the adult zebrafish heart regenerates spontaneously after injury and has been used to study the mechanisms of cardiac repair. However, no zebrafish model is available that mimics ischemic injury in mammalian heart. We developed and characterized zebrafish cardiac injury induced by hypoxia/reox...

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Autores principales: Parente, Valeria, Balasso, Serena, Pompilio, Giulio, Verduci, Lorena, Colombo, Gualtiero I., Milano, Giuseppina, Guerrini, Uliano, Squadroni, Lidia, Cotelli, Franco, Pozzoli, Ombretta, Capogrossi, Maurizio C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3547061/
https://www.ncbi.nlm.nih.gov/pubmed/23341992
http://dx.doi.org/10.1371/journal.pone.0053748
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author Parente, Valeria
Balasso, Serena
Pompilio, Giulio
Verduci, Lorena
Colombo, Gualtiero I.
Milano, Giuseppina
Guerrini, Uliano
Squadroni, Lidia
Cotelli, Franco
Pozzoli, Ombretta
Capogrossi, Maurizio C.
author_facet Parente, Valeria
Balasso, Serena
Pompilio, Giulio
Verduci, Lorena
Colombo, Gualtiero I.
Milano, Giuseppina
Guerrini, Uliano
Squadroni, Lidia
Cotelli, Franco
Pozzoli, Ombretta
Capogrossi, Maurizio C.
author_sort Parente, Valeria
collection PubMed
description AIMS: the adult zebrafish heart regenerates spontaneously after injury and has been used to study the mechanisms of cardiac repair. However, no zebrafish model is available that mimics ischemic injury in mammalian heart. We developed and characterized zebrafish cardiac injury induced by hypoxia/reoxygenation (H/R) and the regeneration that followed it. METHODS AND RESULTS: adult zebrafish were kept either in hypoxic (H) or normoxic control (C) water for 15 min; thereafter fishes were returned to C water. Within 2–6 hours (h) after reoxygenation there was evidence of cardiac oxidative stress by dihydroethidium fluorescence and protein nitrosylation, as well as of inflammation. We used Tg(cmlc2:nucDsRed) transgenic zebrafish to identify myocardial cell nuclei. Cardiomyocyte apoptosis and necrosis were evidenced by TUNEL and Acridine Orange (AO) staining, respectively; 18 h after H/R, 9.9±2.6% of myocardial cell nuclei were TUNEL(+) and 15.0±2.5% were AO(+). At the 30-day (d) time point myocardial cell death was back to baseline (n = 3 at each time point). We evaluated cardiomyocyte proliferation by Phospho Histone H3 (pHH3) or Proliferating Cell Nuclear Antigen (PCNA) expression. Cardiomyocyte proliferation was apparent 18–24 h after H/R, it achieved its peak 3–7d later, and was back to baseline at 30d. 7d after H/R 17.4±2.3% of all cardiomyocytes were pHH3(+) and 7.4±0.6% were PCNA(+) (n = 3 at each time point). Cardiac function was assessed by 2D-echocardiography and Ventricular Diastolic and Systolic Areas were used to compute Fractional Area Change (FAC). FAC decreased from 29.3±2.0% in normoxia to 16.4±1.8% at 18 h after H/R; one month later ventricular function was back to baseline (n = 12 at each time point). CONCLUSIONS: zebrafish exposed to H/R exhibit evidence of cardiac oxidative stress and inflammation, myocardial cell death and proliferation. The initial decrease in ventricular function is followed by full recovery. This model more closely mimics reperfusion injury in mammals than other cardiac injury models.
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spelling pubmed-35470612013-01-22 Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart Parente, Valeria Balasso, Serena Pompilio, Giulio Verduci, Lorena Colombo, Gualtiero I. Milano, Giuseppina Guerrini, Uliano Squadroni, Lidia Cotelli, Franco Pozzoli, Ombretta Capogrossi, Maurizio C. PLoS One Research Article AIMS: the adult zebrafish heart regenerates spontaneously after injury and has been used to study the mechanisms of cardiac repair. However, no zebrafish model is available that mimics ischemic injury in mammalian heart. We developed and characterized zebrafish cardiac injury induced by hypoxia/reoxygenation (H/R) and the regeneration that followed it. METHODS AND RESULTS: adult zebrafish were kept either in hypoxic (H) or normoxic control (C) water for 15 min; thereafter fishes were returned to C water. Within 2–6 hours (h) after reoxygenation there was evidence of cardiac oxidative stress by dihydroethidium fluorescence and protein nitrosylation, as well as of inflammation. We used Tg(cmlc2:nucDsRed) transgenic zebrafish to identify myocardial cell nuclei. Cardiomyocyte apoptosis and necrosis were evidenced by TUNEL and Acridine Orange (AO) staining, respectively; 18 h after H/R, 9.9±2.6% of myocardial cell nuclei were TUNEL(+) and 15.0±2.5% were AO(+). At the 30-day (d) time point myocardial cell death was back to baseline (n = 3 at each time point). We evaluated cardiomyocyte proliferation by Phospho Histone H3 (pHH3) or Proliferating Cell Nuclear Antigen (PCNA) expression. Cardiomyocyte proliferation was apparent 18–24 h after H/R, it achieved its peak 3–7d later, and was back to baseline at 30d. 7d after H/R 17.4±2.3% of all cardiomyocytes were pHH3(+) and 7.4±0.6% were PCNA(+) (n = 3 at each time point). Cardiac function was assessed by 2D-echocardiography and Ventricular Diastolic and Systolic Areas were used to compute Fractional Area Change (FAC). FAC decreased from 29.3±2.0% in normoxia to 16.4±1.8% at 18 h after H/R; one month later ventricular function was back to baseline (n = 12 at each time point). CONCLUSIONS: zebrafish exposed to H/R exhibit evidence of cardiac oxidative stress and inflammation, myocardial cell death and proliferation. The initial decrease in ventricular function is followed by full recovery. This model more closely mimics reperfusion injury in mammals than other cardiac injury models. Public Library of Science 2013-01-16 /pmc/articles/PMC3547061/ /pubmed/23341992 http://dx.doi.org/10.1371/journal.pone.0053748 Text en © 2013 Parente 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
Parente, Valeria
Balasso, Serena
Pompilio, Giulio
Verduci, Lorena
Colombo, Gualtiero I.
Milano, Giuseppina
Guerrini, Uliano
Squadroni, Lidia
Cotelli, Franco
Pozzoli, Ombretta
Capogrossi, Maurizio C.
Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart
title Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart
title_full Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart
title_fullStr Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart
title_full_unstemmed Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart
title_short Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart
title_sort hypoxia/reoxygenation cardiac injury and regeneration in zebrafish adult heart
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3547061/
https://www.ncbi.nlm.nih.gov/pubmed/23341992
http://dx.doi.org/10.1371/journal.pone.0053748
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