Swelling-induced and depolarization-induced C1-channels in normal and cystic fibrosis epithelial cells.
Cl- currents induced by cell swelling were characterized at the whole cell and single-channel levels in primary cultures of normal and cystic fibrosis (CF) epithelial cells and in the T84 cell line. Currents recorded in normal and CF cells were indistinguishable. At 22-24 degrees C with isotonic CsCl in the pipette, initial whole cell outward current density at 100 mV in unswollen cells was 2-4 pA/pF. The current density increased with time during whole cell recording up to 100 pA/pF in isotonic solutions and up to 200 pA/pF in a hypotonic bath, though values typically ranged between 10 and 70 pA/pF. Currents were outwardly rectifying, active at negative voltages, started to inactivate above approximately 40 mV, and were blocked by 4,4'-dinitrostilbene-2,2'-disulfonic acid (DNDS). Single Cl- channels (approximately 50 pS near 0 mV) with an outwardly rectifying current-voltage relation were recorded in cell-attached and outside-out patches from swollen cells. The channels were mostly open at negative voltages and inactivated at positive voltages with a voltage dependence similar to the whole cell currents. Channel activity decreased rapidly (channel rundown) after seal formation. After swelling-induced channel activity had ceased, outwardly rectifying, depolarization-induced Cl- channels (ORDIC channels) were activated in some patches. The swelling-induced and ORDIC single-channel currents were similar, but some consistent differences were observed. ORDIC channels were often closed at resting voltages (-70 to -50 mV), while swelling-induced channels were always open in this voltage range. In addition, ORDIC channels started to inactivate at more positive voltages (approximately 90 vs. approximately 50 mV), rectified more, and had smaller conductances (approximately 25 pS near 0 mV), shorter mean open durations (approximately 70 vs. approximately 350 ms), and more open-channel noise than swelling-induced channels. The two types of currents might arise from separate channel proteins or from a single channel molecule in different states.
Solc
C K
CK
Cystic Fibrosis Research Laboratory, Stanford University, California 94305-2130.
Wine
J J
JJ
D018118:Chloride Channels; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D004553:Electric Conductivity; D004594:Electrophysiology; D004848:Epithelium / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000502:physiology; D012016:Reference Values; D013267:Stilbenes / Q000494:pharmacology; D013545:Sweat Glands / Q000378:metabolism* / Q000473:pathology
D018118:Chloride Channels; D008565:Membrane Proteins; D013267:Stilbenes; C022306:4,4'-dinitro-2,2'-stilbenedisulfonic acid