Does the small conductance Ca2+-activated K+ current ISK flow under physiological conditions in rabbit and human atrial isolated cardiomyocytes?

Abstract

BackgroundThe small conductance Ca2+-activated K+ current (ISK) is a potential therapeutic target for treating atrial fibrillation. AimTo clarify, in rabbit and human atrial cardiomyocytes, the intracellular [Ca2+]-sensitivity of ISK, and its contribution to action potential (AP) repolarisation, under physiological conditions. MethodsWhole-cell-patch clamp, fluorescence microscopy: to record ion currents, APs and [Ca2+]i; 35–37°C. ResultsIn rabbit atrial myocytes, 0.5 mM Ba2+ (positive control) significantly decreased whole-cell current, from −12.8 to −4.9 pA/pF (P < 0.05, n = 17 cells, 8 rabbits). By contrast, the ISK blocker apamin (100 nM) had no effect on whole-cell current, at any set [Ca2+]i (∼100–450 nM). The ISK blocker ICAGEN (1 μM: ≥2 x IC50) also had no effect on current over this [Ca2+]i range. In human atrial myocytes, neither 1 μM ICAGEN (at [Ca2+]i ∼ 100–450 nM), nor 100 nM apamin ([Ca2+]i ∼ 250 nM) affected whole-cell current (5–10 cells, 3–5 patients/group). APs were significantly prolonged (at APD30 and APD70) by 2 mM 4-aminopyridine (positive control) in rabbit atrial myocytes, but 1 μM ICAGEN had no effect on APDs, versus either pre-ICAGEN or time-matched controls. High concentration (10 μM) ICAGEN (potentially ISK-non-selective) moderately increased APD70 and APD90, by 5 and 26 ms, respectively. In human atrial myocytes, 1 μM ICAGEN had no effect on APD30–90, whether stimulated at 1, 2 or 3 Hz (6–9 cells, 2–4 patients/rate). ConclusionISK does not flow in human or rabbit atrial cardiomyocytes with [Ca2+]i set within the global average diastolic-systolic range, nor during APs stimulated at physiological or supra-physiological (≤3 Hz) rates.