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Characterization and functional role of rapid- and slow-activating delayed rectifier K+ currents in atrioventricular node cells of guinea pigs

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Abstract

The atrioventricular (AV) node is the only conduction pathway where electrical impulse can pass from atria to ventricles and exhibits spontaneous automaticity. This study examined the function of the rapid- and slow-activating delayed rectifier K+ currents (IKr and IKs) in the regulation of AV node automaticity. Isolated AV node cells from guinea pigs were current- and voltage-clamped to record the action potentials and the IKr and IKs current. The expression of IKr or IKs was confirmed in the AV node cells by immunocytochemistry, and the positive signals of both channels were localized mainly on the cell membrane. The basal spontaneous automaticity was equally reduced by E4031 and HMR-1556, selective blockers of IKr and IKs, respectively. The nonselective β-adrenoceptor agonist isoproterenol markedly increased the firing rate of action potentials. In the presence of isoproterenol, the firing rate of action potentials was more effectively reduced by the IKs inhibitor HMR-1556 than by the IKr inhibitor E4031. Both E4031 and HMR-1556 prolonged the action potential duration and depolarized the maximum diastolic potential under basal and β-adrenoceptor–stimulated conditions. IKr was not significantly influenced by β-adrenoceptor stimulation, but IKs was concentration-dependently enhanced by isoproterenol (EC50: 15 nM), with a significant negative voltage shift in the channel activation. These findings suggest that both the IKr and IKs channels might exert similar effects on regulating the repolarization process of AV node action potentials under basal conditions; however, when the β-adrenoceptor is activated, IKs modulation may become more important.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Abbreviations

AV node:

Atrioventricular node

SA node:

Sinoatrial node

I K :

The delayed rectifier K+ current

I Kr :

The rapid component of delayed rectifier K+ current

I Ks :

The slow component of delayed rectifier K+ current

I Ca,L :

L-type Ca2+ current

I f :

Hyperpolarization-activated inward current

PKA:

Protein kinase A

MDP:

Maximal diastolic potential

max dV·dt−1 :

Maximum depolarizing velocity

APD90 :

Action potential duration at 90% repolarization

DMSO:

Dimethylsulfoxide

DAPI:

4′-6′-diamidino-2-phenylindole

PBS:

Phosphate-buffered saline

TBST:

Tris-buffered saline with 0.1% Tween-20

C m :

Cell membrane capacitance

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Funding

This study was supported by the Japan Society for the Promotion of Science (Tokyo, Japan) KAKENHI Grant Numbers 17K11050 (to Akiko Kojima) and 17K08536 (to Hiroshi Matsuura).

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Authors and Affiliations

  1. Department of Anesthesiology, Shiga University of Medical Science, Otsu, Shiga, 520-2192, Japan

    Mayumi Yuasa, Akiko Kojima & Hirotoshi Kitagawa

  2. Department of Physiology, Shiga University of Medical Science, Otsu, Shiga, 520-2192, Japan

    Xinya Mi, Wei-Guang Ding, Mariko Omatsu-Kanbe & Hiroshi Matsuura

Authors
  1. Mayumi Yuasa
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  2. Akiko Kojima
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  3. Xinya Mi
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Contributions

MY and HM participated in the research design; MY, XM, MO, and HM conducted the experiments; MY, AK, HK, W-GD, and HM performed the data analysis; and MY, W-GD, AK, and HM wrote the manuscript. All authors revised the final version of the manuscript and approved its submission.

Corresponding author

Correspondence to Wei-Guang Ding.

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Ethics approval

All animal care and experimental procedures were conducted in accordance with the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996) and were approved by the institution’s Animal Care and Use Committee of Shiga University of Medical Science. The studies involving animals were reported in accordance with the ARRIVE guidelines for reporting experiments involving animals [30].

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Supplementary information

Supplemental Figure 1

The effects of the sequential application of HMR-1556, E4031 and a mixture of HMR-1556 and E4031 on AV node automaticity under basal or β-adrenoceptor-activated conditions in one series of experiments. a Action potentials recorded under basal conditions (1), and the application of 1 μM HMR-1556 (2), HMR-1556 washout (3), the application of 5 μM E4031 (4), a mixture of E4031 and HMR-1556 (5) under basal conditions. b Summarized data for the reduction in the firing rate induced by E4031, HMR-1556 and by a mixture of E4031 and HMR-1556, under basal conditions (n = 11, N = 6). *, P < 0.05 in comparison to control. c Action potentials recorded under the application of 0.01 μM isoproterenol (6), and the application of 1 μM HMR-1556 (7), HMR-1556 washout (8), 5 μM E4031 (9), a mixture of E4031 and HMR-1556 (10), under β-adrenoceptor-activated conditions. d Summarized data for the reduction in the firing rate induced by E4031, HMR-1556 and by a mixture of E4031 and HMR-1556 in the presence of isoproterenol (n = 16, N = 7). *, P < 0.05 in comparison to the application of isoproterenol alone. (PNG 713 kb)

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Yuasa, M., Kojima, A., Mi, X. et al. Characterization and functional role of rapid- and slow-activating delayed rectifier K+ currents in atrioventricular node cells of guinea pigs. Pflugers Arch - Eur J Physiol 473, 1885–1898 (2021). https://doi.org/10.1007/s00424-021-02617-z

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