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Inadvertent phosphorylation of survival kinases in isolated perfused hearts: a word of caution

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Abstract

The isolated perfused heart is an important model in cardiovascular research. We hypothesized that the perfusion procedure per se will phosphorylate some protein kinases important in pre- and postconditioning. Isolated hearts were Langendorff-perfused for 20 min with or without an intraventricular balloon (rats and mice), or in the working heart mode (mice) and compared to non-perfused controls with respect to protein phosphorylation. Rat hearts were also perfused for 20 and 50 min in the Langendorff mode to investigate the effect of perfusion time on phosphorylation. Western blot analysis showed that perfusion per se induced a massive phosphorylation of ERK 1/2, P38-MAPK, JNK, AMPK, but decreased phosphorylation of AKT in the isolated rat and mouse heart. However, during ongoing perfusion the phosphorylation of these kinases was reduced. Langendorff-perfusion without the intraventricular balloon caused less phosphorylation of ERK 1/2, P38-MAPK and JNK, but had no effect on AMPK. In working hearts phosphorylation of kinases was similar to that of Langendorff-perfused hearts without the balloon. Our findings indicate that excising, handling and perfusion induce a time dependent phosphorylation of stress kinases. The presence of the intraventricular balloon caused the strongest phosphorylation, thus Langendorff-perfused hearts might be partly protected by the perfusion procedure if stress kinases are protective in pre- and postconditioning. This might explain conflicting results obtained with different models of both pre- and postconditioning, and the isolated heart might in some situations be suboptimal for such studies.

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Acknowledgments

The technical assistance by May Britt Dahl is gratefully acknowledged. Disclosures: None. Sources of funding: This study has been supported by the University of Oslo (JV), Ullevål University. Hospital (KOS and JV), Eastern Norway Regional Health Authority (JV) and Norwegian National Health Association (JV).

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Authors and Affiliations

  1. Dept. of Surgery, Ullevål University Hospital, Oslo, Norway

    Kåre-Olav Stensløkken & Jarle Vaage

  2. Institute of Experimental Medical Research, Ullevål University Hospital, 0407, Oslo, Norway

    Kåre-Olav Stensløkken, Arkady Rutkovskiy, Mari-Liis Kaljusto & Jarle Vaage

  3. Dept. of Cardiothoracic Surgery, Ullevål University Hospital, Oslo, Norway

    Mari-Liis Kaljusto

  4. Dept. of Medical Physiology, Institute of Medical Biology, University of Tromsø, Tromsø, Norway

    Anne Dragøy Hafstad & Terje Steinar Larsen

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  1. Kåre-Olav Stensløkken
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  2. Arkady Rutkovskiy
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Correspondence to Kåre-Olav Stensløkken.

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Returned for 1. Revision: 4 September 2008 1. Revision received: 20 October 2008

Returned for 2. Revision: 31 October 2008 2. Revision received: 5 December 2008

Returned for 3. Revision: 15 December 2008 3. Revision received: 6 January 2009

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Stensløkken, KO., Rutkovskiy, A., Kaljusto, ML. et al. Inadvertent phosphorylation of survival kinases in isolated perfused hearts: a word of caution. Basic Res Cardiol 104, 412–423 (2009). https://doi.org/10.1007/s00395-009-0780-1

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