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Effect of MANT-nucleotides on L-type calcium currents in murine cardiomyocytes

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Abstract

Membranous adenylyl cyclases play a major role in G-protein-coupled receptor signalling and regulate various cellular responses, such as cardiac contraction. Cardiac apoptosis and development of cardiac dysfunction is prevented in mice lacking AC 5, a predominant isoform in the heart. In the search for a potent and selective AC 5 inhibitor, we recently identified 2′(3′)-methylanthraniloyl-inosine-5′-triphosphate(MANT-ITP) as the most potent AC 5 inhibitor with a K i of 13 nM. Therefore, AC inhibition of MANT-ITP was assessed in ventricular cardiomyocytes and compared to three other MANT-nucleotides to evaluate its effect on cardiac signalling. Basal and isoproterenol-induced L-type calcium currents (I Ca,L) in murine ventricular cardiomyocytes were recorded by whole-cell patch-clamp technique, using four different MANT-nucleotides. The effects of the MANT-nucleotides on I Ca,L were unexpectedly complex. All MANT-nucleotides exhibited an inhibitory effect on basal I Ca,L. Additionally, several MANT-nucleotides, i.e., MANT-ITPγS, MANT-ATP, and MANT-ITP, caused a strong initial increase in basal I Ca,L within the first 2.5 min that appeared to be unrelated to AC 5 inhibition. However, we detected a significant reduction on isoproterenol-induced I Ca,L with MANT-ITP, supporting the notion that AC 5 plays an important role in agonist-stimulated activation of I Ca,L. Collectively, MANT-nucleotides are useful tools for the characterization of recombinant ACs, for fluorescence studies and crystallography, but in intact cardiomyocytes, caution must be exerted since MANT-nucleotides apparently possess additional effects than AC 5 inhibition, limiting their usefulness as tools for intact cell studies.

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Acknowledgment

This work was supported by a graduate scholarship of the Elite Network of Bavaria (to M.H.) and Deutsche Forschungsgemeinschaft grant 529/5-2 (to R.S.). We are very grateful to the reviewers for their constructive and helpful comments.

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, University of Regensburg, Regensburg, Germany

    Melanie Hübner

  2. Department of Pharmacology, University of Cologne, Gleueler Str. 24, 50931, Cologne, Germany

    Sara Dizayee, Jan Matthes & Stefan Herzig

  3. Institute for Pharmacology, Medical School of Hannover, Hannover, Germany

    Roland Seifert

Authors
  1. Melanie Hübner
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  2. Sara Dizayee
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  3. Jan Matthes
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  4. Roland Seifert
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  5. Stefan Herzig
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Correspondence to Stefan Herzig.

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Hübner, M., Dizayee, S., Matthes, J. et al. Effect of MANT-nucleotides on L-type calcium currents in murine cardiomyocytes. Naunyn-Schmiedeberg's Arch Pharmacol 383, 573–583 (2011). https://doi.org/10.1007/s00210-011-0626-x

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  • DOI: https://doi.org/10.1007/s00210-011-0626-x

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