Functional role of inward rectifier current in heart probed by Kir2.1 overexpression and dominant-negative suppression.
The inward rectifier current I(K1) is tightly regulated regionally within the heart, downregulated in heart failure, and genetically suppressed in Andersen syndrome. We used in vivo viral gene transfer to dissect the role of I(K1) in cardiac repolarization and maintenance of the resting membrane potential (RMP) in guinea pig ventricular myocytes. Kir2.1 overexpression boosted Ba(2+)-sensitive I(K1) by more than 100% (at -50mV), significantly shortened action potential durations (APDs), accelerated phase 3 repolarization, and hyperpolarized RMP compared with control cells (nongreen cells from the same hearts and green cells from GFP-transduced hearts). The dominant-negative Kir2.1AAA reduced I(K1) by 50-90%; those cells with less than 80% reduction of I(K1) exhibited prolonged APDs, decelerated phase 3 repolarization, and depolarization of the RMP. Further reduction of I(K1) resulted in a pacemaker phenotype, as previously described. ECGs revealed a 7.7% +/- 0.9% shortening of the heart rate-corrected QT interval (QTc interval) in Kir2.1-transduced animals (n = 4) and a 16.7% +/- 1.8% prolongation of the QTc interval (n = 3) in Kir2.1AAA-transduced animals 72 hours after gene delivery compared with immediate postoperative recordings. Thus, I(K1) is essential for establishing the distinctive electrical phenotype of the ventricular myocyte: rapid terminal repolarization to a stable and polarized resting potential. Additionally, the long-QT phenotype seen in Andersen syndrome is a direct consequence of dominant-negative suppression of Kir2 channel function.
Institute of Molecular Cardiobiology, Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Junichiro
J
Miake
Eduardo
E
Marbán
H Bradley
HB
Nuss
D000200:Action Potentials / Q000502:physiology; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002460:Cell Line; D002469:Cell Separation; D004562:Electrocardiography; D005260:Female; D018014:Gene Transfer Techniques; D005799:Genes, Dominant; D006168:Guinea Pigs; D006321:Heart / Q000502:physiology*; D006801:Humans; D009154:Mutation; D009206:Myocardium / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000502:physiology; D018408:Patch-Clamp Techniques; D010641:Phenotype; D024661:Potassium Channels, Inwardly Rectifying / Q000096:biosynthesis* / Q000235:genetics*; D014161:Transduction, Genetic
D024661:Potassium Channels, Inwardly Rectifying