Inhibition of Ca2+-dependent K+ channels in rat carotid body type I cells by protein kinase C

J Physiol. 1998 Nov 1;512 ( Pt 3)(Pt 3):743-50. doi: 10.1111/j.1469-7793.1998.743bd.x.

Abstract

1. Whole-cell patch clamp recordings were used to investigate the effects of protein kinase C (PKC) activation on K+ and Ca2+ currents in type I cells isolated from the rat carotid body. 2. Pretreatment of cells for 10 min at 37 C with 4alpha-phorbol 12,13-didecanoate (4alpha-PDD, 200 nM), a phorbol ester which does not activate PKC, did not affect K+ current density as compared with cells pretreated with vehicle alone. By contrast, identical pretreatment with 200 nM 12-O-teradecanoylphorbol-13-acetate (TPA, a PKC activator) dramatically reduced K+ current density in type I cells. This effect was prevented by co-incubation of cells with the PKC inhibitor bisindolylmaleimide (BIM, 3 microM). 3. The sensitivity of K+ currents to inhibition by 200 microM Cd2+ (indicative of the presence of Ca2+-dependent K+ channels) was markedly reduced in TPA-treated cells as compared with sham-treated cells, cells treated with 4alpha-PDD, and cells treated with both TPA and BIM. Cd2+-resistant K+ current densities were of similar magnitude in all four groups of cells, as were the input resistances determined over the voltage range -100 mV to -50 mV. 4. Ca2+ channel current density was not significantly different in type I cells pretreated with 200 nM 4alpha-PDD as compared with cells treated with the same concentration of TPA. 5. The degree of inhibition of K+ currents caused by hypoxia (Po2 15-20 mmHg) was unaltered by pretreatment of cells with 3 microM BIM. 6. The resting membrane potential of cells pretreated with TPA was depolarized as compared with controls, and the Ca2+-dependent K+ channel inhibitor iberiotoxin (20 nM) failed to depolarize these cells further. 7. Our results suggest that activation of PKC causes a marked, selective inhibition of Ca2+-dependent K+ currents in type I carotid body cells, but that PKC activation is unlikely to account for inhibition of these channels by acute hypoxia.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / physiology*
  • Carotid Body / cytology
  • Carotid Body / drug effects
  • Carotid Body / physiology*
  • Electric Stimulation
  • Electrophysiology
  • Enzyme Activation / drug effects
  • Enzyme Activation / physiology
  • Enzyme Inhibitors / pharmacology
  • In Vitro Techniques
  • Indoles / pharmacology
  • Maleimides / pharmacology
  • Membrane Potentials / drug effects
  • Membrane Potentials / physiology
  • Patch-Clamp Techniques
  • Phorbol Esters / pharmacology
  • Potassium Channel Blockers*
  • Potassium Channels / physiology
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / metabolism
  • Protein Kinase C / physiology*
  • Rats
  • Tetradecanoylphorbol Acetate / pharmacology

Substances

  • Enzyme Inhibitors
  • Indoles
  • Maleimides
  • Phorbol Esters
  • Potassium Channel Blockers
  • Potassium Channels
  • phorbol-12,13-didecanoate
  • Protein Kinase C
  • bisindolylmaleimide
  • Tetradecanoylphorbol Acetate
  • Calcium