CaMKII modulates the cardiac transient outward K+ current through their association in non-caveolar membrane rafts

Background/aims: To test whether the physiological regulation of the cardiac Kv4 channels by the Ca2+/calmodulin-dependent protein kinase II (CaMKII) is restricted to lipid rafts and whether the interactions observed in rat cardiomyocytes also occur in the human ventricle. Methods: Ventricular myocy...

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Autores: Alday, Aintzane, Ahyayauch, Hasna, Fernández López, Víctor, Echeazarra Escudero, Leyre, Urrutia Iñiguez, Janire, Casis Sáenz, Oscar, Gallego Muñoz, Mónica
Tipo de documento: artigo
Data de publicação:2020
País:España
Recursos:Universidad del País Vasco
Repositório:Addi. Archivo Digital para la Docencia y la Investigación
OAI Identifier:oai:addi.ehu.eus:10810/65082
Acesso em linha:http://hdl.handle.net/10810/65082
Access Level:Acceso aberto
Palavra-chave:calmodulin
ito
Kv4.3
Kv4.2
lipid rafts
caveola
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oai_identifier_str oai:addi.ehu.eus:10810/65082
network_acronym_str ES
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repository_id_str
spelling CaMKII modulates the cardiac transient outward K+ current through their association in non-caveolar membrane raftsAlday, AintzaneAhyayauch, HasnaFernández López, VíctorEcheazarra Escudero, LeyreUrrutia Iñiguez, JanireCasis Sáenz, OscarGallego Muñoz, MónicacalmodulinitoKv4.3Kv4.2lipid raftscaveolaBackground/aims: To test whether the physiological regulation of the cardiac Kv4 channels by the Ca2+/calmodulin-dependent protein kinase II (CaMKII) is restricted to lipid rafts and whether the interactions observed in rat cardiomyocytes also occur in the human ventricle. Methods: Ventricular myocytes were freshly isolated from Sprague-Dawley rats. Ito was recorded by the whole-cell Patch-Clamp technique. Membrane rafts were isolated by centrifugation in a discontinuous sucrose density gradient. The presence of the proteins of interest was analysed by western blot. Immunogold staining and electron microscopy of heart vibrosections was performed to localize Kv4.2/Kv4.3 and CaMKII proteins. Protein-protein interactions were determined by co-immunoprecipitation experiments in rat and human ventricular mycoytes. Results: Patch-Clamp recordings in control conditions and after lipid raft or caveolae disruption show that the CaMKII-Kv4 channel complex must associate in non-caveolar lipid rafts to be functional. Separation in density gradients, co-immunoprecipitation and electron microscopy show that there are two Kv4 channel populations: one located in caveolae, that is CaMKII independent, and another one located in planar membrane rafts, which is bound to CaMKII. Conclusion: CaMKII regulates only the Kv4 channel population located in non-caveolar lipid rafts. Thus, the regulation of cardiac Kv4 channels in rat and human ventricle depends on their subcellular localization.This work was supported by grants from the MICINN (SAF2007-61159) and the Basque Government (PIBA2018-58; GIC18/150). V.F.-L. is a predoctoral fellow of the University of the Basque Country UPV/EHU. The monoclonal antibodies anti-Kv4.2 and Kv4.3 were developed by and/or obtained from the UC Davis/NIH NeuroMab Facility, supported by NIH grant U24NS050606 and maintained by the Department of Neurobiology, Physiology and Behaviour, College of Biological Sciences, University of California, Davis, CA 95616.University of Duisburg-Essen202420242020info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10810/65082reponame:Addi. Archivo Digital para la Docencia y la Investigacióninstname:Universidad del País VascoIngléshttps://www.cellphysiolbiochem.com/Articles/000203/info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-nd/4.0/deed.en© 2020 The Author(s). Published by Cell Physiol Biochem Press GmbH&Co. This article is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 Interna- tional License (CC BY-NC-ND). Usage and distribution for commercial purposes as well as any distribution of modified material requires written permission.oai:addi.ehu.eus:10810/650822026-06-18T09:23:17Z
dc.title.none.fl_str_mv CaMKII modulates the cardiac transient outward K+ current through their association in non-caveolar membrane rafts
title CaMKII modulates the cardiac transient outward K+ current through their association in non-caveolar membrane rafts
spellingShingle CaMKII modulates the cardiac transient outward K+ current through their association in non-caveolar membrane rafts
Alday, Aintzane
calmodulin
ito
Kv4.3
Kv4.2
lipid rafts
caveola
title_short CaMKII modulates the cardiac transient outward K+ current through their association in non-caveolar membrane rafts
title_full CaMKII modulates the cardiac transient outward K+ current through their association in non-caveolar membrane rafts
title_fullStr CaMKII modulates the cardiac transient outward K+ current through their association in non-caveolar membrane rafts
title_full_unstemmed CaMKII modulates the cardiac transient outward K+ current through their association in non-caveolar membrane rafts
title_sort CaMKII modulates the cardiac transient outward K+ current through their association in non-caveolar membrane rafts
dc.creator.none.fl_str_mv Alday, Aintzane
Ahyayauch, Hasna
Fernández López, Víctor
Echeazarra Escudero, Leyre
Urrutia Iñiguez, Janire
Casis Sáenz, Oscar
Gallego Muñoz, Mónica
author Alday, Aintzane
author_facet Alday, Aintzane
Ahyayauch, Hasna
Fernández López, Víctor
Echeazarra Escudero, Leyre
Urrutia Iñiguez, Janire
Casis Sáenz, Oscar
Gallego Muñoz, Mónica
author_role author
author2 Ahyayauch, Hasna
Fernández López, Víctor
Echeazarra Escudero, Leyre
Urrutia Iñiguez, Janire
Casis Sáenz, Oscar
Gallego Muñoz, Mónica
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv calmodulin
ito
Kv4.3
Kv4.2
lipid rafts
caveola
topic calmodulin
ito
Kv4.3
Kv4.2
lipid rafts
caveola
description Background/aims: To test whether the physiological regulation of the cardiac Kv4 channels by the Ca2+/calmodulin-dependent protein kinase II (CaMKII) is restricted to lipid rafts and whether the interactions observed in rat cardiomyocytes also occur in the human ventricle. Methods: Ventricular myocytes were freshly isolated from Sprague-Dawley rats. Ito was recorded by the whole-cell Patch-Clamp technique. Membrane rafts were isolated by centrifugation in a discontinuous sucrose density gradient. The presence of the proteins of interest was analysed by western blot. Immunogold staining and electron microscopy of heart vibrosections was performed to localize Kv4.2/Kv4.3 and CaMKII proteins. Protein-protein interactions were determined by co-immunoprecipitation experiments in rat and human ventricular mycoytes. Results: Patch-Clamp recordings in control conditions and after lipid raft or caveolae disruption show that the CaMKII-Kv4 channel complex must associate in non-caveolar lipid rafts to be functional. Separation in density gradients, co-immunoprecipitation and electron microscopy show that there are two Kv4 channel populations: one located in caveolae, that is CaMKII independent, and another one located in planar membrane rafts, which is bound to CaMKII. Conclusion: CaMKII regulates only the Kv4 channel population located in non-caveolar lipid rafts. Thus, the regulation of cardiac Kv4 channels in rat and human ventricle depends on their subcellular localization.
publishDate 2020
dc.date.none.fl_str_mv 2020
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10810/65082
url http://hdl.handle.net/10810/65082
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv https://www.cellphysiolbiochem.com/Articles/000203/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/deed.en
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/deed.en
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv University of Duisburg-Essen
publisher.none.fl_str_mv University of Duisburg-Essen
dc.source.none.fl_str_mv reponame:Addi. Archivo Digital para la Docencia y la Investigación
instname:Universidad del País Vasco
instname_str Universidad del País Vasco
reponame_str Addi. Archivo Digital para la Docencia y la Investigación
collection Addi. Archivo Digital para la Docencia y la Investigación
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.300719

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