Recapitulation of an Ion Channel IV Curve Using Frequency Components

INTRODUCTION: Presently, there are no established methods to measure multiple ion channel types simultaneously and decompose the measured current into portions attributable to each channel type. This study demonstrates how impedance spectroscopy may be used to identify specific frequencies that highly correlate with the steady state current amplitude measured during voltage clamp experiments. The method involves inserting a noise function containing specific frequencies into the voltage step protocol. In the work presented, a model cell is used to demonstrate that no high correlations are introduced by the voltage clamp circuitry, and also that the noise function itself does not introduce any high correlations when no ion channels are present. This validation is necessary before the technique can be applied to preparations containing ion channels. The purpose of the protocol presented is to demonstrate how to characterize the frequency response of a single ion channel type to a noise function. Once specific frequencies have been identified in an individual channel type, they can be used to reproduce the steady state current voltage (IV) curve. Frequencies that highly correlate with one channel type and minimally correlate with other channel types may then be used to estimate the current contribution of multiple channel types measured simultaneously

Electrophysiologic effects of the IK1 inhibitor PA-6 are modulated by extracellular potassium in isolated guinea pig hearts

The pentamidine analog PA‐6 was developed as a specific inward rectifier potassium current (I(K) (1)) antagonist, because established inhibitors either lack specificity or have side effects that prohibit their use in vivo. We previously demonstrated that BaCl(2), an established I(K) (1) inhibitor, could prolong action potential duration (APD) and increase cardiac conduction velocity (CV). However, few studies have addressed whether targeted I(K) (1) inhibition similarly affects ventricular electrophysiology. The aim of this study was to determine the effects of PA‐6 on cardiac repolarization and conduction in Langendorff‐perfused guinea pig hearts. PA‐6 (200 nm) or vehicle was perfused into ex‐vivo guinea pig hearts for 60 min. Hearts were optically mapped with di‐4‐ANEPPS to quantify CV and APD at 90% repolarization (APD (90)). Ventricular APD (90) was significantly prolonged in hearts treated with PA‐6 (115 ± 2% of baseline; P < 0.05), but not vehicle (105 ± 2% of baseline). PA‐6 slightly, but significantly, increased transverse CV by 7%. PA‐6 significantly prolonged APD (90) during hypokalemia (2 mmol/L [K+](o)), although to a lesser degree than observed at 4.56 mmol/L [K+](o). In contrast, the effect of PA‐6 on CV was more pronounced during hypokalemia, where transverse CV with PA‐6 (24 ± 2 cm/sec) was significantly faster than with vehicle (13 ± 3 cm/sec, P < 0.05). These results show that under normokalemic conditions, PA‐6 significantly prolonged APD (90), whereas its effect on CV was modest. During hypokalemia, PA‐6 prolonged APD (90) to a lesser degree, but profoundly increased CV. Thus, in intact guinea pig hearts, the electrophysiologic effects of the I(K) (1) inhibitor, PA‐6, are [K+](o)‐dependent