Distinct voltage-dependent gating behaviours of a swelling-activated chloride current in human epithelial cells.
1. The swelling-activated chloride current is critical in the homeostatic regulatory volume decrease (RVD) of both excitable and non-excitable cells. Although not activated by voltage, it displays kinetic behaviour similar to voltage-gated Shaker-type potassium currents. We have studied the voltage-dependent properties of this current in single T84 human cell line epithelial cells using whole-cell patch clamp methodology. 2. An external anion permeability sequence of I- > Cl- > methanesulphonate (MeSO3-) was observed for the swelling-activated current. Extracellular application of the chloride channel blocker DIDS (100 microM) resulted in approximately 50% block of the current in a voltage-dependent manner. 3. At positive membrane potentials, the swelling-activated chloride current undergoes time-dependent inactivation. Following such inactivation, recovery of both the inward and outward components of the macroscopic current was found to be voltage dependent. The time constants describing these two individual recovery processes were identical over a range of membrane potentials. In addition, the magnitude of current recovery was directly dependent upon the degree prior inactivation of current at positive voltage. 4. We further observed that the swelling-activated current undergoes a form of steady-state, voltage-dependent inactivation that appears to differ from the inactivation observed at positive potentials. This steady-state inactivation occurred over the physiological voltage range, with a membrane potential at half-maximal inactivation (V1/2) of -72 mV, and differed from the time-dependent inactivation observed at positive membrane potentials, which occurred with a V1/2 of 40 mV. These observations demonstrate two distinct forms of voltage-dependent inactivation, probably reflecting two separate gating processes at the level of the channel. 5. These latter properties are thus anticipated to regulate voltage-dependent chloride efflux under cell swelling conditions and further influence RVD and membrane excitability in cells generating action potentials.
Braun
A P
AP
Department of Neurobiology, Stanford University School of Medicine, CA 94305-5401, USA.
Schulman
H
H
D002460:Cell Line; D048429:Cell Size; D018118:Chloride Channels / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism*; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006801:Humans; D015640:Ion Channel Gating; D017136:Ion Transport; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008564:Membrane Potentials; D018408:Patch-Clamp Techniques; D014882:Water-Electrolyte Balance
D018118:Chloride Channels; D002712:Chlorides; D008274:Magnesium