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Protein kinase/phosphatase balance mediates the effects of increased late sodium current on ventricular calcium cycling

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Abstract

Increased late sodium current (late INa) is an important arrhythmogenic trigger in cardiac disease. It prolongs cardiac action potential and leads to an increased SR Ca2+ leak. This study investigates the contribution of Ca2+/Calmodulin-dependent kinase II (CaMKII), protein kinase A (PKA) and conversely acting protein phosphatases 1 and 2A (PP1, PP2A) to this subcellular crosstalk. Augmentation of late INa (ATX-II) in murine cardiomyocytes led to an increase of diastolic Ca2+ spark frequency and amplitudes of Ca2+ transients but did not affect SR Ca2+ load. Interestingly, inhibition of both, CaMKII and PKA, attenuated the late INa-dependent induction of the SR Ca2+ leak. PKA inhibition additionally reduced the amplitudes of systolic Ca2+ transients. FRET-measurements revealed increased levels of cAMP upon late INa augmentation, which could be prevented by simultaneous inhibition of Na+/Ca2+-exchanger (NCX) suggesting that PKA is activated by Ca2+-dependent cAMP-production. Whereas inhibition of PP2A showed no effect on late INa-dependent alterations of Ca2+ cycling, additional inhibition of PP1 further increased the SR Ca2+ leak. In line with this, selective activation of PP1 yielded a strong reduction of the late INa-induced SR Ca2+ leak and did not affect systolic Ca2+ release. This study indicates that phosphatase/kinase-balance is perturbed upon increased Na+ influx leading to disruption of ventricular Ca2+ cycling via CaMKII- and PKA-dependent pathways. Importantly, an activation of PP1 at RyR2 may represent a promising new toehold to counteract pathologically increased kinase activity.

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Acknowledgements

We gratefully acknowledge the technical assistance of K.-C. Hansing and T. Schulte. THF is funded by the Deutsche Forschungsgemeinschaft (DFG) through the SFB 1002 (A11). SS is supported by the Marga und Walter Boll-Stiftung through a research grant. MK is funded by the European Research Council (ERC) through a starting Grant (# 336567).

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Author notes

  1. Jörg Eiringhaus and Jonas Herting contributed equally to this work.

Authors and Affiliations

  1. Abt. Kardiologie und Pneumologie/Herzzentrum, Georg-August-Universität Göttingen, Robert-Koch-Str. 40, 37075, Göttingen, Germany

    Jörg Eiringhaus, Jonas Herting, Felix Schatter, Julia Sprenger, Markus Zabel, Gerd Hasenfuss, Samuel Sossalla & Thomas H. Fischer

  2. Abt. Kardiologie und Angiologie, Medizinische Hochschule Hannover, Hannover, Germany

    Jörg Eiringhaus

  3. Institut für Experimentelle Herz-Kreislaufforschung, Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany

    Viacheslav O. Nikolaev

  4. European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, Germany

    Yansong Wang & Maja Köhn

  5. Centre for Biological Signalling Studies (BIOSS) and Faculty of Biology, University of Freiburg, Freiburg, Germany

    Maja Köhn

  6. Institut für Pharmakologie und Toxikologie, Technische Universität Dresden, Dresden, Germany

    Ali El-Armouche

  7. Klinik und Poliklinik für Innere Medizin II, Universitätsklinikum Regensburg, Regensburg, Germany

    Samuel Sossalla

  8. Medizinische Klinik II, Kardiologie, Angiologie, Pneumologie, Klinikum Coburg, Coburg, Germany

    Thomas H. Fischer

  9. Deutsches Zentrum für Herz-Kreislauf Forschung (DZHK), Standort Göttingen, Göttingen, Germany

    Jörg Eiringhaus, Jonas Herting, Felix Schatter, Markus Zabel, Gerd Hasenfuss, Samuel Sossalla & Thomas H. Fischer

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  1. Jörg Eiringhaus
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Eiringhaus, J., Herting, J., Schatter, F. et al. Protein kinase/phosphatase balance mediates the effects of increased late sodium current on ventricular calcium cycling. Basic Res Cardiol 114, 13 (2019). https://doi.org/10.1007/s00395-019-0720-7

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