Characterization of inwardly rectifying K(+) conductance across the basolateral membrane of rat tracheal epithelia

Biochem Biophys Res Commun. 2001 Nov 9;288(4):914-20. doi: 10.1006/bbrc.2001.5831.

Abstract

The rat primary cultured-airway monolayer has been an excellent model for deciphering the ion channel after nystatin permeabilization of its basolateral or apical membrane. Inwardly rectifying K(+) currents were characterized across the basolateral membrane in symmetrical HCO(-)(3)-free high K(+) Ringer's solution (125 mM) in this study. The potency of K(+) channel inhibitors against K(+) conductance was Ba(2+) (IC(50) = 5 microM) > Cs(+) (IC(50) = 2 mM) >> glybenclamide (IC(50) > 5 mM) >> TEA (IC(50) >> 100 mM). The application of basolateral Cs(+) changed K(+) conductance into an oscillating current, and its frequency (holding voltage = -100 mV) increased with increase in concentration of basolateral Cs(+) (0.05-5 mM) and in degree of hyperpolarization. Addition of basolateral Cs(+) blocked inward current strongly at -100 mV and hardly at all at -60 mV, giving a sharp curvature to the I-V relation of the IRK current. RT-PCR, Western blotting, and immunohistochemical analyses showed that Kir2.1 might be present in basolateral membrane of tracheal epithelia and plasma membrane of pulmonary alveolar cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Calcium / metabolism
  • Calcium / pharmacology
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism*
  • Cell Membrane Permeability
  • Cell Polarity
  • Cells, Cultured
  • Cloning, Molecular
  • Electric Conductivity
  • Humans
  • Immunohistochemistry
  • Ion Channel Gating / drug effects
  • Membrane Potentials / drug effects
  • Molecular Sequence Data
  • Myocardium / metabolism
  • Nasal Mucosa / cytology
  • Nasal Mucosa / drug effects
  • Nasal Mucosa / metabolism
  • Potassium / metabolism*
  • Potassium Channel Blockers / pharmacology
  • Potassium Channels / chemistry
  • Potassium Channels / genetics
  • Potassium Channels / metabolism*
  • Potassium Channels, Inwardly Rectifying*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Respiratory Mucosa / cytology
  • Respiratory Mucosa / drug effects
  • Respiratory Mucosa / metabolism*
  • Respiratory System / cytology
  • Respiratory System / metabolism
  • Trachea / cytology
  • Trachea / drug effects
  • Trachea / metabolism*

Substances

  • Potassium Channel Blockers
  • Potassium Channels
  • Potassium Channels, Inwardly Rectifying
  • RNA, Messenger
  • Potassium
  • Calcium