Transient receptor potential melastatin 4 channel inhibitor 9-phenanthrol inhibits K+ but not Ca2+ currents in canine ventricular myocytes

Abstract

The role of transient receptor potential melastatin 4 (TRPM4) channels has been frequently tested using their inhibitor 9-phenanthrol in various cardiac preparations; however, the selectivity of the compound is uncertain. Therefore, in the present study, the concentration-dependent effects of 9-phenanthrol on major ionic currents were studied in canine isolated ventricular cells using whole-cell configuration of the patch-clamp technique and 10 mM BAPTA-containing pipette solution to prevent the Ca²⁺-dependent activation of TRPM4 channels. Transient outward (I_to1), rapid delayed rectifier (I_Kr), and inward rectifier (I_K1) K⁺ currents were suppressed by 10 and 30 μM 9-phenanthrol with the blocking potency for I_K1 < I_Kr < I_to1 and partial reversibility. L-type Ca²⁺ current was not affected up to the concentration of 30 μM. In addition, a steady outward current was detected at voltages positive to -40 mV in 9-phenanthrol, which was larger at more positive voltages and larger 9-phenanthrol concentrations. Action potentials were recorded using microelectrodes. Maximal rate of depolarization, phase-1 repolarization, and terminal repolarization were decreased and the plateau potential was depressed by 9-phenanthrol (3-30 μM), congruently with the observed alterations of ionic currents. Significant action potential prolongation was observed by 9-phenanthrol in the majority of the studied cells, but only at 30 μM concentration. In conclusion, 9-phenanthrol is not selective to TRPM4 channels in canine ventricular myocardium; therefore, its application as a TRPM4 blocker can be appropriate only in expression systems but not in native cardiac cells.

Keywords: 9-phenanthrol; 9-phénanthrol; TRPM4 channels; canaux TRPM4; canine myocytes; cardiac action potentials; cardiac ionic currents; cardiomyocytes ventriculaires canins; courants ioniques cardiaques; potentiels d’action cardiaques.