Small-Conductance Calcium-Activated Potassium Current in Normal Rabbit Cardiac Purkinje Cells

J Am Heart Assoc. 2017 May 26;6(6):e005471. doi: 10.1161/JAHA.117.005471.

Abstract

Background: Purkinje cells (PCs) are important in cardiac arrhythmogenesis. Whether small-conductance calcium-activated potassium (SK) channels are present in PCs remains unclear. We tested the hypotheses that subtype 2 SK (SK2) channel proteins and apamin-sensitive SK currents are abundantly present in PCs.

Methods and results: We studied 25 normal rabbit ventricles, including 13 patch-clamp studies, 4 for Western blotting, and 8 for immunohistochemical staining. Transmembrane action potentials were recorded in current-clamp mode using the perforated-patch technique. For PCs, the apamin (100 nmol/L) significantly prolonged action potential duration measured to 80% repolarization by an average of 10.4 ms (95% CI, 0.11-20.72) (n=9, P=0.047). Voltage-clamp study showed that apamin-sensitive SK current density was significantly larger in PCs compared with ventricular myocytes at potentials ≥0 mV. Western blotting of SK2 expression showed that the SK2 protein expression in the midmyocardium was 58% (P=0.028) and the epicardium was 50% (P=0.018) of that in the pseudotendons. Immunostaining of SK2 protein showed that PCs stained stronger than ventricular myocytes. Confocal microscope study showed SK2 protein was distributed to the periphery of the PCs.

Conclusions: SK2 proteins are more abundantly present in the PCs than in the ventricular myocytes of normal rabbit ventricles. Apamin-sensitive SK current is important in ventricular repolarization of normal PCs.

Keywords: action potential; apamin; potassium channels; repolarization.

Publication types

  • Comparative Study

MeSH terms

  • Action Potentials
  • Animals
  • Apamin / pharmacology
  • Blotting, Western
  • Heart Ventricles / cytology
  • Heart Ventricles / drug effects
  • Heart Ventricles / metabolism*
  • Immunohistochemistry
  • Microscopy, Confocal
  • Myocytes, Cardiac / metabolism
  • Patch-Clamp Techniques
  • Potassium / metabolism*
  • Purkinje Fibers / drug effects
  • Purkinje Fibers / metabolism*
  • Rabbits
  • Small-Conductance Calcium-Activated Potassium Channels / drug effects
  • Small-Conductance Calcium-Activated Potassium Channels / metabolism*
  • Time Factors

Substances

  • Small-Conductance Calcium-Activated Potassium Channels
  • Apamin
  • Potassium