Involvement of anion channel(s) in the modulation of the transient outward K(+) channel in rat ventricular myocytes.

The cardiac Ca(2+)-independent transient outward K(+) current (I(to)), a major repolarizing ionic current, is markedly affected by Cl(-) substitution and anion channel blockers. We reexplored the mechanism of the action of anions on I(to) by using whole cell patch-clamp in single isolated rat cardiac ventricular myocytes. The transient outward current was sensitive to blockade by 4-aminopyridine (4-AP) and was abolished by Cs(+) substitution for intracellular K(+). Replacement of most of the extracellular Cl(-) with less permeant anions, aspartate (Asp(-)) and glutamate (Glu(-)), markedly suppressed the current. Removal of external Na(+) or stabilization of F-actin with phalloidin did not significantly affect the inhibitory action of less permeant anions on I(to). In contrast, the permeant Cl(-) substitute Br(-) did not markedly affect the current, whereas F(-) substitution for Cl(-) induced a slight inhibition. The I(to) elicited during Br(-) substitution for Cl(-) was also sensitive to blockade by 4-AP. The ability of Cl(-) substitutes to induce rightward shifts of the steady-state inactivation curve of I(to) was in the following sequence: NO(3)(-) > Cl(-) approximately Br(-) > gluconate(-) > Glu(-) > Asp(-). Depolymerization of actin filaments with cytochalasin D (CytD) induced an effect on the steady-state inactivation of I(to) similar to that of less permeant anions. Fluorescent phalloidin staining experiments revealed that CytD-pretreatment significantly decreased the intensity of FITC-phalloidin staining of F-actin, whereas Asp(-) substitution for Cl(-) was without significant effect on the intensity. These results suggest that the I(to) channel is modulated by anion channel(s), in which the actin cytoskeleton may be implicated.