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BIOSOLVIX ARCHIVE특허 및 논문

Spontaneous inward currents reflecting oscillatory activation of Na/Ca2 exchangers in human embryonic stem cell-derived cardiom

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2024-02-01 10:02
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Choi SW, Lee HA, Moon SH, Park SJ, Kim HJ, Kim KS, Zhang YH, Youm JB, Kim SJ. Spontaneous inward currents reflecting oscillatory activation of Na/Ca2 exchangers in human embryonic stem cell-derived cardiomyocytes. Pflugers Arch. 2016 Jul;468(7):1295.

Abstract



Na+/Ca2+ exchanger current (INCX) triggered by spontaneous Ca2+ release from sarcoplasmic reticulum (SR) has been suggested as one of the cardiac pacemaker mechanisms (“Ca2+ clock model”). In human embryonic stem cell-derived cardiomyocytes (hESC-CMs) showing spontaneous action potentials (APs), we found that substantial population (35 %) showed regular oscillation of inward currents (SICs) in nystatin-perforated voltage clamp between −40 and 40 mV (−80 ± 10.6 pA, at −20 mV). SICs were similarly observed between nodal, atrial, and ventricular hESC-CMs. Oscillations of [Ca2+]i synchronized with SICs were observed under voltage clamp. SICs were eliminated by lowering [Ca2+]e, L-type Ca2+ channel (VOCCL) blocker (nifedipine, 10 µM), ryanodine receptor (RyR) agonist (caffeine, 10 mM), or NCX inhibitor (1 µM SN-6 and 10 µM KB-R7943). Plasma membrane expression of NCX1 was confirmed using immunofluorescence confocal microcopy. Both caffeine and SN-6 slowed the pacemaker potential but did not abolish the AP generation. The inhibitors of funny current (3 µM ivabradine) or voltage-gated K+ channel currents (1 µM E4031 and 10 µM chromanol-293B) also did not abolish but slowed the pacemaker potential. In a computational model of cardiac pacemaker by Maltsev and Lakatta (2009), after modifying the spatial distribution of RyR, VOCCL, and NCX by using our multiparameter adjust algorithm, we could successfully reproduce spontaneous SR Ca2+ release and SICs under voltage clamp. It was proposed that, under the membrane depolarization activating VOCCL, oscillatory Ca2+ releases via RyR induce sharp increases in subsarcolemmal [Ca2+]i and inward INCX (SICs). Since the hESC-CMs without SICs still showed spontaneous APs, the putative “Ca2+ clock” would provide a redundant pacemaker or augmenting mechanism in hESC-CMs.


 

Keywords: #Ca2+ release #Embryonic stem cell #Heart #NCX #Pacemaker current

 

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