Contribution of chloride shifts to the fade of gamma-aminobutyric acid-gated currents in frog dorsal root ganglion cells

Abstract

1. The contribution of Cl- redistribution to the decay phase of the GABA (gamma-aminobutyric acid) response was investigated in isolated frog sensory neurones, using a suction-pipette technique which allows for internal perfusion under conditions of voltage clamp. 2. In neurones perfused with 120 mM [Cl-]i and [Cl-]o at driving forces (delta VH) of less than 15 mV, no shift of GABA equilibrium potential (EGABA) occurred during a continuous application of GABA, at various concentrations. However, increases of delta VH towards negative or positive potentials over 15 mV induced EGABA shifts. 3. The degree of EGABA shift was governed by the total amount of Cl- flux across the soma membrane, an event which depends upon delta VH, GABA concentration and drug application time. 4. The time-dependent EGABA shift due to Cl- redistribution during GABA application induced a current run-down resulting from a decreased Cl- gradient and a diminished Cl- conductance (gCl), the latter brought about by a drop in the intracellular ionic density of Cl-. 5. The EGABA shift during a continuous GABA application was also affected by [Cl-]i; e.g. the shift more readily occurred at lower [Cl-]i. 6. In neurones perfused with internal and external solutions containing 120 mM-Cl- at a delta VH of less than 10 mV, the change of gCl occurred with no shift of EGABA during the continuous application of GABA at concentrations over 6 x 10(-5) M, thereby indicating a 'real' GABA receptor desensitization. The desensitization depended solely upon the agonist concentrations but not upon the amount of ICl. Under these conditions, the time course of recovery from GABA desensitization was estimated. The decrease of gCl at the desensitization phase was a single exponential. 7. At a delta VH greater than 15 mV, therefore, the decay of ICl induced by GABA concentrations over 6 x 10(-6) M consists of the sum of both the 'real' GABA receptor desensitization and the current run-down brought about by Cl- shifts. The gCl at the current decay phase consisted of a double exponential. In the present experiments we chose experimental conditions with which Cl- shift become negligible. 8. The 'pure' GABA receptor desensitization during a continuous application of GABA developed rapidly at GABA concentrations over 10(-5) M. The speed of desensitization was facilitated by increasing the magnitude of desensitization.