Intracellular chloride modulates A-type potassium currents in astrocytes.
Application of the GABA(A) receptor agonist muscimol to astrocytes in situ or in vitro results in a receptor-mediated Cl(-) current with a concomitant block of outward K(+) currents. The effect on K(+) current is largely selective for the inactivating A-type current. Parallel experiments with various Cl(-) pipette concentrations show a significant reduction in A-type current under low Cl(-) conditions with minimal effect on delayed current. In addition, lower Cl(-) conditions caused a depolarizing shift of steady-state inactivation (V(1/2), -68 to -57 mV) and activation (V(1/2), -5.8 to 34 mV) kinetics of A-type current only. Cl(-) had no effect on the time course of inactivation or reactivation kinetics, suggesting the Cl(-)-mediated effect is largely on activation kinetics, indirectly affecting steady-state inactivation. Muscimol application to astrocytes under perforated patch control (gramicidin) displayed a similar block of A-type current to that of conventional whole cell patch at 40 or 20 mM pipette Cl(-) concentrations. With barium application under perforated patch conditions, the study of muscimol-mediated Cl(-) current in isolation of the effect on K(+) currents was possible. This allowed estimation of intracellular Cl(-) concentration from receptor current reversal information. The average intracellular Cl(-) concentration was found to be 29 +/- 3.2 mM. The effect on activation kinetics and lack of effect on time course of inactivation or reactivation suggest that intracellular anion concentrations have an effect on the K(+) channel voltage sensor region. Cl(-) may modulate K(+) currents by altering membrane field potentials surrounding K(+) channel proteins.
Department of Physiology, University of Saskatchewan, Saskatoon, Canada.
Lane K
LK
Bekar
Wolfgang
W
Walz
D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D002712:Chlorides / Q000494:pharmacology / Q000502:physiology*; D058785:GABA-A Receptor Agonists; D007424:Intracellular Fluid / Q000187:drug effects / Q000502:physiology*; D009118:Muscimol / Q000494:pharmacology; D024642:Potassium Channels, Voltage-Gated / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D011963:Receptors, GABA-A / Q000502:physiology
D002712:Chlorides; D058785:GABA-A Receptor Agonists; D024642:Potassium Channels, Voltage-Gated; D011963:Receptors, GABA-A; D009118:Muscimol