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Ischemia and Reperfusion Induce Differential Expression of Calpastatin and Its Homologue High Molecular Weight Calmodulin-Binding Protein in Murine Cardiomyocytes
In the heart, calpastatin (Calp) and its homologue high molecular weight calmodulin-binding protein (HMWCaMBP) regulate calpains (Calpn) by inhibition. A rise in intracellular myocardial Ca(2+) during cardiac ischemia activates Calpn thereby causing damage to myocardial proteins, which leads to myoc...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4259361/ https://www.ncbi.nlm.nih.gov/pubmed/25486053 http://dx.doi.org/10.1371/journal.pone.0114653 |
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author | Parameswaran, Sreejit Sharma, Rajendra K. |
author_facet | Parameswaran, Sreejit Sharma, Rajendra K. |
author_sort | Parameswaran, Sreejit |
collection | PubMed |
description | In the heart, calpastatin (Calp) and its homologue high molecular weight calmodulin-binding protein (HMWCaMBP) regulate calpains (Calpn) by inhibition. A rise in intracellular myocardial Ca(2+) during cardiac ischemia activates Calpn thereby causing damage to myocardial proteins, which leads to myocyte death and consequently to loss of myocardial structure and function. The present study aims to elucidate expression of Calp and HMWCaMBP with respect to Calpn during induced ischemia and reperfusion in primary murine cardiomyocyte cultures. Ischemia and subsequently reperfusion was induced in ∼80% confluent cultures of neonatal murine cardiomyocytes (NMCC). Flow cytometric analysis (FACS) has been used for analyzing protein expression concurrently with viability. Confocal fluorescent microscopy was used to observe protein localization. We observed that ischemia induces increased expression of Calp, HMWCaMBP and Calpn. Calpn expressing NMCC on co-expressing Calp survived ischemic induction compared to NMCC co-expressing HMWCaMBP. Similarly, living cells expressed Calp in contrast to dead cells which expressed HMWCaMBP following reperfusion. A significant difference in the expression of Calp and its homologue HMWCaMBP was observed in localization studies during ischemia. The current study adds to the existing knowledge that HMWCaMBP could be a putative isoform of Calp. NMCC on co-expressing Calp and Calpn-1 survived ischemic and reperfusion inductions compared to NMCC co-expressing HMWCaMBP and Calpn-1. A significant difference in expression of Calp and HMWCaMBP was observed in localization studies during ischemia. |
format | Online Article Text |
id | pubmed-4259361 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-42593612014-12-15 Ischemia and Reperfusion Induce Differential Expression of Calpastatin and Its Homologue High Molecular Weight Calmodulin-Binding Protein in Murine Cardiomyocytes Parameswaran, Sreejit Sharma, Rajendra K. PLoS One Research Article In the heart, calpastatin (Calp) and its homologue high molecular weight calmodulin-binding protein (HMWCaMBP) regulate calpains (Calpn) by inhibition. A rise in intracellular myocardial Ca(2+) during cardiac ischemia activates Calpn thereby causing damage to myocardial proteins, which leads to myocyte death and consequently to loss of myocardial structure and function. The present study aims to elucidate expression of Calp and HMWCaMBP with respect to Calpn during induced ischemia and reperfusion in primary murine cardiomyocyte cultures. Ischemia and subsequently reperfusion was induced in ∼80% confluent cultures of neonatal murine cardiomyocytes (NMCC). Flow cytometric analysis (FACS) has been used for analyzing protein expression concurrently with viability. Confocal fluorescent microscopy was used to observe protein localization. We observed that ischemia induces increased expression of Calp, HMWCaMBP and Calpn. Calpn expressing NMCC on co-expressing Calp survived ischemic induction compared to NMCC co-expressing HMWCaMBP. Similarly, living cells expressed Calp in contrast to dead cells which expressed HMWCaMBP following reperfusion. A significant difference in the expression of Calp and its homologue HMWCaMBP was observed in localization studies during ischemia. The current study adds to the existing knowledge that HMWCaMBP could be a putative isoform of Calp. NMCC on co-expressing Calp and Calpn-1 survived ischemic and reperfusion inductions compared to NMCC co-expressing HMWCaMBP and Calpn-1. A significant difference in expression of Calp and HMWCaMBP was observed in localization studies during ischemia. Public Library of Science 2014-12-08 /pmc/articles/PMC4259361/ /pubmed/25486053 http://dx.doi.org/10.1371/journal.pone.0114653 Text en © 2014 Parameswaran, Sharma 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 Parameswaran, Sreejit Sharma, Rajendra K. Ischemia and Reperfusion Induce Differential Expression of Calpastatin and Its Homologue High Molecular Weight Calmodulin-Binding Protein in Murine Cardiomyocytes |
title | Ischemia and Reperfusion Induce Differential Expression of Calpastatin and Its Homologue High Molecular Weight Calmodulin-Binding Protein in Murine Cardiomyocytes |
title_full | Ischemia and Reperfusion Induce Differential Expression of Calpastatin and Its Homologue High Molecular Weight Calmodulin-Binding Protein in Murine Cardiomyocytes |
title_fullStr | Ischemia and Reperfusion Induce Differential Expression of Calpastatin and Its Homologue High Molecular Weight Calmodulin-Binding Protein in Murine Cardiomyocytes |
title_full_unstemmed | Ischemia and Reperfusion Induce Differential Expression of Calpastatin and Its Homologue High Molecular Weight Calmodulin-Binding Protein in Murine Cardiomyocytes |
title_short | Ischemia and Reperfusion Induce Differential Expression of Calpastatin and Its Homologue High Molecular Weight Calmodulin-Binding Protein in Murine Cardiomyocytes |
title_sort | ischemia and reperfusion induce differential expression of calpastatin and its homologue high molecular weight calmodulin-binding protein in murine cardiomyocytes |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4259361/ https://www.ncbi.nlm.nih.gov/pubmed/25486053 http://dx.doi.org/10.1371/journal.pone.0114653 |
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