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01-05-2011 | Original Contribution

Evidence for an intracellular localization of the adenosine A_2B receptor in rat cardiomyocytes

Authors: Karina Grube, Julia Rüdebusch, Zhelong Xu, Thomas Böckenholt, Carmen Methner, Tobias Müller, Friederike Cuello, Katrin Zimmermann, Xiulan Yang, Stephan B. Felix, Michael V. Cohen, James M. Downey, Thomas Krieg

Published in: Basic Research in Cardiology | Issue 3/2011

Abstract

Protection achieved by ischemic preconditioning is dependent on A_2B adenosine receptors (A_2BAR) in rabbit and mouse hearts and, predictably, an A_2BAR agonist protects them. But it is controversial whether cardiomyocytes themselves actually express A_2BAR. The present study tested whether A_2BAR could be demonstrated on rat cardiomyocytes. Isolated rat hearts experienced 30 min of ischemia and 120 min of reperfusion. The highly selective, cell-permeant A_2BAR agonist BAY60-6583 (500 nM) infused at reperfusion reduced infarct size from 40.4 ± 2.0% of the risk zone in control hearts to 19.9 ± 2.8% indicating that A_2BAR are protective in rat heart as well. Furthermore, BAY60-6583 reduced calcium-induced mitochondrial permeability transition in isolated rat cardiomyocytes. A_2BAR protein could be demonstrated in isolated cardiomyocytes by western blotting. In addition, message for A_2BAR was found in individual cardiomyocytes using quantitative RT-PCR. Surprisingly, immunofluorescence microscopy did not show A_2BAR on the cardiomyocyte’s sarcolemma but rather at intracellular sites. Co-staining with MitoTracker Red in isolated cardiomyocytes revealed A_2BAR are localized to mitochondria. Western blot analysis of a mitochondrial fraction from either rat heart biopsies or isolated cardiomyocytes revealed a strong A_2BAR band. Thus, the present study demonstrates that activation of A_2BAR is strongly cardioprotective in rat heart and suppresses transition pores in isolated cardiomyocytes, and A_2BAR are expressed in individual cardiomyocytes. However, surprisingly, A_2BAR are present in or near mitochondria rather than on the sarcolemma as are other adenosine receptors. Because A_2BAR signaling is thought to result in inhibition of mitochondrial transition pores, this convenient location may be important.

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Title: Evidence for an intracellular localization of the adenosine A2B receptor in rat cardiomyocytes
Authors: Karina Grube
Julia Rüdebusch
Zhelong Xu
Thomas Böckenholt
Carmen Methner
Tobias Müller
Friederike Cuello
Katrin Zimmermann
Xiulan Yang
Stephan B. Felix
Michael V. Cohen
James M. Downey
Thomas Krieg
Publication date: 01-05-2011
Publisher: Springer-Verlag
Published in: Basic Research in Cardiology / Issue 3/2011
Print ISSN: 0300-8428
Electronic ISSN: 1435-1803
DOI: https://doi.org/10.1007/s00395-011-0151-6

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