Abstract：
OBJECTIVE:The spontaneous action potential of isolated sinoatrial node (SAN) cells is regulated by a coupled-clock system of two clocks: the calcium clock and membrane clock. However, it remains unclear whether calcium clock inhibitors have a direct effect on the membrane clock. The purpose of this study was to investigate the direct effect of cyclopiazonic acid (CPA), a selective calcium clock inhibitor, on the function of the membrane clock of SAN cells. METHODS:at SAN cells were isolated by trypsinization and identified based on morphology and electrophysiology. If and HCN currents were recorded via patch clamp technique. The expression of the HCN channel protein was determined by Western blotting analysis. RESULTS:The diastolic depolarization rate of spontaneous action potentials and the current densities of If were reduced by exposure to 10 μM CPA. The inhibitory effect of CPA was concentration-dependent with an IC50 value of 16.3 μM and a Hill coefficient of 0.98. The effect of CPA on If current was also time-dependent, and the If current amplitude was partially restored after washout. Furthermore, the steady-state activation curve of the If current was shifted to a negative potential, indicating that channel activation slowed down. Finally, the protein expression of HCN4 in HEK293 cells was markedly downregulated by CPA. CONCLUSIONS:These results indicate that the direct inhibition effect of CPA on the If current in SAN cells is both concentration- and time-dependent. The underlying mechanisms may involve slowing down steady-state activation and the downregulation of pacemaker channel protein expression.
journal_name
Neurosciencejournal_title
Neuroscienceauthors
Chen Q,Chen JQ,Zhu PL,Wu HL,Xie XT,Dong Y,Xiang GJ,Chen MY,Li Y,Zhang JCdoi
10.1016/j.neuroscience.2020.01.018subject
Has Abstractpub_date
2020-05-01 00:00:00pages
230-240eissn
0306-4522issn
1873-7544pii
S0306-4522(20)30031-2journal_volume
433pub_type
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