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ROS Production via P2Y(1)-PKC-NOX2 Is Triggered by Extracellular ATP after Electrical Stimulation of Skeletal Muscle Cells
During exercise, skeletal muscle produces reactive oxygen species (ROS) via NADPH oxidase (NOX2) while inducing cellular adaptations associated with contractile activity. The signals involved in this mechanism are still a matter of study. ATP is released from skeletal muscle during electrical stimul...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4460042/ https://www.ncbi.nlm.nih.gov/pubmed/26053483 http://dx.doi.org/10.1371/journal.pone.0129882 |
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author | Díaz-Vegas, Alexis Campos, Cristian A. Contreras-Ferrat, Ariel Casas, Mariana Buvinic, Sonja Jaimovich, Enrique Espinosa, Alejandra |
author_facet | Díaz-Vegas, Alexis Campos, Cristian A. Contreras-Ferrat, Ariel Casas, Mariana Buvinic, Sonja Jaimovich, Enrique Espinosa, Alejandra |
author_sort | Díaz-Vegas, Alexis |
collection | PubMed |
description | During exercise, skeletal muscle produces reactive oxygen species (ROS) via NADPH oxidase (NOX2) while inducing cellular adaptations associated with contractile activity. The signals involved in this mechanism are still a matter of study. ATP is released from skeletal muscle during electrical stimulation and can autocrinely signal through purinergic receptors; we searched for an influence of this signal in ROS production. The aim of this work was to characterize ROS production induced by electrical stimulation and extracellular ATP. ROS production was measured using two alternative probes; chloromethyl-2,7- dichlorodihydrofluorescein diacetate or electroporation to express the hydrogen peroxide-sensitive protein Hyper. Electrical stimulation (ES) triggered a transient ROS increase in muscle fibers which was mimicked by extracellular ATP and was prevented by both carbenoxolone and suramin; antagonists of pannexin channel and purinergic receptors respectively. In addition, transient ROS increase was prevented by apyrase, an ecto-nucleotidase. MRS2365, a P2Y(1) receptor agonist, induced a large signal while UTPyS (P2Y(2) agonist) elicited a much smaller signal, similar to the one seen when using ATP plus MRS2179, an antagonist of P2Y(1). Protein kinase C (PKC) inhibitors also blocked ES-induced ROS production. Our results indicate that physiological levels of electrical stimulation induce ROS production in skeletal muscle cells through release of extracellular ATP and activation of P2Y1 receptors. Use of selective NOX2 and PKC inhibitors suggests that ROS production induced by ES or extracellular ATP is mediated by NOX2 activated by PKC. |
format | Online Article Text |
id | pubmed-4460042 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-44600422015-06-16 ROS Production via P2Y(1)-PKC-NOX2 Is Triggered by Extracellular ATP after Electrical Stimulation of Skeletal Muscle Cells Díaz-Vegas, Alexis Campos, Cristian A. Contreras-Ferrat, Ariel Casas, Mariana Buvinic, Sonja Jaimovich, Enrique Espinosa, Alejandra PLoS One Research Article During exercise, skeletal muscle produces reactive oxygen species (ROS) via NADPH oxidase (NOX2) while inducing cellular adaptations associated with contractile activity. The signals involved in this mechanism are still a matter of study. ATP is released from skeletal muscle during electrical stimulation and can autocrinely signal through purinergic receptors; we searched for an influence of this signal in ROS production. The aim of this work was to characterize ROS production induced by electrical stimulation and extracellular ATP. ROS production was measured using two alternative probes; chloromethyl-2,7- dichlorodihydrofluorescein diacetate or electroporation to express the hydrogen peroxide-sensitive protein Hyper. Electrical stimulation (ES) triggered a transient ROS increase in muscle fibers which was mimicked by extracellular ATP and was prevented by both carbenoxolone and suramin; antagonists of pannexin channel and purinergic receptors respectively. In addition, transient ROS increase was prevented by apyrase, an ecto-nucleotidase. MRS2365, a P2Y(1) receptor agonist, induced a large signal while UTPyS (P2Y(2) agonist) elicited a much smaller signal, similar to the one seen when using ATP plus MRS2179, an antagonist of P2Y(1). Protein kinase C (PKC) inhibitors also blocked ES-induced ROS production. Our results indicate that physiological levels of electrical stimulation induce ROS production in skeletal muscle cells through release of extracellular ATP and activation of P2Y1 receptors. Use of selective NOX2 and PKC inhibitors suggests that ROS production induced by ES or extracellular ATP is mediated by NOX2 activated by PKC. Public Library of Science 2015-06-08 /pmc/articles/PMC4460042/ /pubmed/26053483 http://dx.doi.org/10.1371/journal.pone.0129882 Text en © 2015 Díaz-Vegas 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 Díaz-Vegas, Alexis Campos, Cristian A. Contreras-Ferrat, Ariel Casas, Mariana Buvinic, Sonja Jaimovich, Enrique Espinosa, Alejandra ROS Production via P2Y(1)-PKC-NOX2 Is Triggered by Extracellular ATP after Electrical Stimulation of Skeletal Muscle Cells |
title | ROS Production via P2Y(1)-PKC-NOX2 Is Triggered by Extracellular ATP after Electrical Stimulation of Skeletal Muscle Cells |
title_full | ROS Production via P2Y(1)-PKC-NOX2 Is Triggered by Extracellular ATP after Electrical Stimulation of Skeletal Muscle Cells |
title_fullStr | ROS Production via P2Y(1)-PKC-NOX2 Is Triggered by Extracellular ATP after Electrical Stimulation of Skeletal Muscle Cells |
title_full_unstemmed | ROS Production via P2Y(1)-PKC-NOX2 Is Triggered by Extracellular ATP after Electrical Stimulation of Skeletal Muscle Cells |
title_short | ROS Production via P2Y(1)-PKC-NOX2 Is Triggered by Extracellular ATP after Electrical Stimulation of Skeletal Muscle Cells |
title_sort | ros production via p2y(1)-pkc-nox2 is triggered by extracellular atp after electrical stimulation of skeletal muscle cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4460042/ https://www.ncbi.nlm.nih.gov/pubmed/26053483 http://dx.doi.org/10.1371/journal.pone.0129882 |
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