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Enrichment of cardiac pacemaker-like cells: neuregulin-1 and cyclic AMP increase I f-current density and connexin 40 mRNA levels in fetal cardiomyocytes

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Abstract

Generation of a large number of cells belonging to the cardiac pacemaker system would constitute an important step towards their utilization as a biological cardiac pacemaker system. The aim of the present study was to identify factors, which might induce transformation of a heterogenous population of fetal cardiomyocytes into cells with a pacemaker-like phenotype. Neuregulin-1 (α- and β-isoform) or the cAMP was added to fresh cell cultures of murine embryonic cardiomyocytes. Quantitative northern blot analysis and flowcytometry were performed to detect the expression of connexins 40, 43 and 45. Patch clamp recordings in the whole cell configuration were performed to determine current density of I f, a characteristic ion current of pacemaker cells. Fetal cardiomyocytes without supplement of neuregulin or cAMP served as control group. Neuregulin and cAMP significantly increased mRNA levels of connexin 40 (Cx-40), a marker of the early differentiating conduction system in mice. On the protein level, flowcytometry revealed no significant differences between treated and untreated groups with regard to the expression of connexins 40, 43 and 45. Treatment with cAMP (11.2 ± 2.24 pA/pF; P < 0.001) and neuregulin-1-β (6.23 ± 1.07 pA/pF; P < 0.001) significantly increased the pacemaker current density compared to control cardiomyocytes (1.76 ± 0.49 pA/pF). Our results indicate that neuregulin-1 and cAMP possess the capacity to cause significant transformation of a mixed population of fetal cardiomyocytes into cardiac pacemaker-like cells as shown by electrophysiology and increase of Cx-40 mRNA. This method may allow the development of a biological cardiac pacemaker system when applied to adult or embryonic stem cells.

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Acknowledgments

This study was supported by the Deutsche Forschungsgemeinschaft (Ho 2146/3-1)

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Authors and Affiliations

  1. Department of Cardiac Surgery, University of Heidelberg, 69120, Heidelberg, Germany

    Arjang Ruhparwar & Matthias Karck

  2. Department of Internal Medicine III, University of Cologne, Kerpener Str. 62, 50937, Cologne, Germany

    Fikret Er & Uta C. Hoppe

  3. Department of Thoracic and Cardiovascular Surgery, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Ulrich Martin, Kristin Radke, Ina Gruh & Axel Haverich

  4. Department of Cardiology and Angiology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Michael Niehaus

  5. Center for Molecular Medicine, University of Cologne (CMMC), Cologne, Germany

    Uta C. Hoppe

Authors
  1. Arjang Ruhparwar
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  2. Fikret Er
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  3. Ulrich Martin
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  4. Kristin Radke
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  5. Ina Gruh
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  6. Michael Niehaus
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  7. Matthias Karck
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  8. Axel Haverich
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  9. Uta C. Hoppe
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Correspondence to Arjang Ruhparwar.

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Ruhparwar, A., Er, F., Martin, U. et al. Enrichment of cardiac pacemaker-like cells: neuregulin-1 and cyclic AMP increase I f-current density and connexin 40 mRNA levels in fetal cardiomyocytes. Med Bio Eng Comput 45, 221–227 (2007). https://doi.org/10.1007/s11517-007-0164-3

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  • DOI: https://doi.org/10.1007/s11517-007-0164-3

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