A volume-sensitive chloride conductance in human colonic cell line T84.
The chloride-secreting colonic cell line, T84, was studied under whole cell patch clamp with Cl as the permeant ion in pipette and bath solutions. Transmembrane current was initially small (approximately 50 pA at +100 mV) but increased steadily to average values of 1-3 nA within 5-10 min. The development of this current was associated with visible cell swelling, either without a shape change or with membrane blebbing. Basal, preswelling current levels were restored by the addition of 50-75 mM sucrose to the bath or when pipette osmolality was reduced by an equivalent amount. These findings suggest that an isosmotic pipette filling solution behaves as if it is hypertonic by approximately 60 mosmol/kgH2O to the bath. Currents traversing the swelling-induced conductance were outwardly rectified and showed activation at hyperpolarizing voltages and inactivation at depolarizing voltages. They were Cl selective because the reversal potential for current flow approached the Cl equilibrium potential when bath [Cl] was varied. Under nonswelling conditions (bath solution, 300 mosmol/kgH2O; pipette solution, 240 mosmol/kgH2O), single-channel steps (approximately 9 pA at +100 mV) could be resolved. The single-channel characteristics were similar to the macroscopic currents recorded from swollen cells, showing inactivation at positive voltages and an outwardly rectified current-voltage relation. Summation of these single-channel events yielded currents that were similar to those from swollen cells, implying that activation of multiple channels with these properties is the basis of the swelling-induced Cl conductance. This volume-sensitive Cl conductance would contribute to a regulatory volume decrease when T84 cells swell. Its relation to the secretory Cl conductance in these cells is unknown.
Worrell
R T
RT
Department of Physiology and Biophysics, University of Alabama, Birmingham 35294.
Butt
A G
AG
Cliff
W H
WH
Frizzell
R A
RA
D002460:Cell Line; D018118:Chloride Channels; D002712:Chlorides / Q000502:physiology*; D003110:Colonic Neoplasms; D004553:Electric Conductivity; D006801:Humans; D007473:Ion Channels / Q000502:physiology*; D007700:Kinetics; D008564:Membrane Potentials; D008565:Membrane Proteins / Q000502:physiology*
D018118:Chloride Channels; D002712:Chlorides; D007473:Ion Channels; D008565:Membrane Proteins