Existence, properties, and functional expression of "Maxi-K"-type, Ca2+-activated K+ channels in short-term cultured hepatocytes

J Cell Physiol. 1997 Apr;171(1):87-94. doi: 10.1002/(SICI)1097-4652(199704)171:1<87::AID-JCP10>3.0.CO;2-L.

Abstract

A large-conductance, Ca2+-activated K+ channel was identified and characterized in embryonic chick hepatocytes using the patch-electrode voltage-clamp technique. The channel conductance was 213 pS in excised patches bathed in symmetrical 145 mM KCI and 1 mM Ca2+. Current-voltage relationships were linear with high K+ on both sides of the membrane but showed constant field rectification as the K+ gradient was increased. The reversal potential shifted 58 mV per 10-fold change in the ratio of external to internal K+. Channel openings occurred at potentials higher than +50 mV in cell-attached patches. The open probability X voltage relationship shifted to more negative potentials in excised, inside-out patches exposed to a solution containing high Ca2+. The voltage sensitivity of the channel was not significantly affected by changes in internal Ca2+ concentration. Conversely, channel gating, reflected in the half-activation potential, shifted 118 mV per 10-fold change in internal Ca2+ at concentrations less than approximately 2 microM, although at higher Ca2+, this parameter was Ca2+ insensitive. Channel open probability in cell-attached patches increased significantly following exposure of the cells to either the Ca2+ ionophore A-23187 or L-alanine, a cell-volume modulator. Channel density increased with time spent in culture from no observations in 10-hr cells, through 13 and 80% of patches in 24-and 48-hr cultured cells, respectively. The implications of delayed functional expression for ion channel studies in acutely dissociated cells is discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcimycin / pharmacology
  • Calcium / metabolism*
  • Cells, Cultured
  • Chickens
  • Ion Channel Gating
  • Ionophores / pharmacology
  • Liver / metabolism*
  • Potassium Channels / metabolism*

Substances

  • Ionophores
  • Potassium Channels
  • Calcimycin
  • Calcium