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Beta-adrenoceptor subtype dependence of chronotropy in mouse embryonic stem cell-derived cardiomyocytes

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Abstract.

Cardiomyocytes derived from embryonic stem cells (ESCM) have potential both as an experimental model for investigating cardiac physiology and as a source for tissue repair. For both reasons it is important to characterise the responses of these cells, and one of the key modulators of contraction is the β-adrenergic system. We therefore undertook a detailed study of the response of the spontaneous beating rate of ESCM to β-adrenoceptor (βAR) stimulation. Embryoid bodies (EBs) were generated from murine ES line E14Tg2a by the hanging drop method, followed by plating. Spontaneously beating areas were seen starting from 9–14 days after differentiation: the experiments described here were performed on EBs between developmental day 19 and 48. Beating cell layers were seeded with charcoal to allow tracking of movement by a video-edge detection system. Experiments were performed in physiological medium containing 1 mM Ca2+ at 37 °C. Isoprenaline (Iso) increased beating rate with an EC50 value of 52 nM. Iso (0.3 µM) increased basal rate from 67 ± 7 beats per minute (bpm) to 138 ± 18 bpm, P < 0.001, n = 22. At earlier developmental time points the response to Iso was not maintained through 5 min exposure; this spontaneous desensitisation only being observed before day 36. A repeat application of Iso after a wash period of 20 min produced reproducible effects on beating rate. Subtype dependence of the βAR response was determined by comparing an initial response with a second in the presence of selective β1- or β2AR antagonists. In the presence of the specific β1AR-blocker CGP 20712A (300 nM) the increase in rate with Iso was reduced from 207 ± 42% of basal to 128 ± 13%, P < 0.01. With the β2AR-blocker ICI 118,551 (50 nM) there was no significant change in Iso response. Exposure to the muscarinic agonist, carbachol (10 µM), inhibited the increase in frequency mediated by isoprenaline, but had mixed stimulatory and inhibitory effects on basal rate. This study extends the characterisation of ESCM as a preparation for studying receptor pharmacology, and indicates that the β1AR is the predominant subtype mediating increases in contraction rate in murine ESCM.

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Authors and Affiliations

  1. NHLI Division, Imperial College London, Dovehouse St., London SW3 6LY, UK

    N. N. Ali, X. Xu, M. Brito-Martins, P. A. Poole-Wilson,  S. E. Harding & S. J. Fuller

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  1. N. N. Ali
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  2. X. Xu
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  4. P. A. Poole-Wilson
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Correspondence to S. E. Harding.

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Ali, N.N., Xu, X., Brito-Martins, M. et al. Beta-adrenoceptor subtype dependence of chronotropy in mouse embryonic stem cell-derived cardiomyocytes. Basic Res Cardiol 99, 382–391 (2004). https://doi.org/10.1007/s00395-004-0484-5

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  • DOI: https://doi.org/10.1007/s00395-004-0484-5

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