Sodium-dependent suppression of gamma aminobutyric-acid-gated chloride currents in internally perfused frog sensory neurons

Abstract

1. The effects of the Na+ electrochemical potential gradient on gamma-aminobutyric acid (GABA)-induced Cl- currents (ICl) in frog sensory neurones were studied, using a suction pipette technique with which internal perfusion can be accomplished under current- and voltage-clamp conditions. 2. Under current clamp, the depolarizing response to GABA decreased in the presence of external Na+. A similar external Na+-dependent reduction in the GABA-induced inward ICl was observed under voltage clamp. The reversal potential of GABA-induced ICl (EGABA) was nearly equal to the Cl- equilibrium potential (ECl), irrespective of the presence or absence of external Na+. 3. Varying the Na+ influx by changing the holding membrane potential (VH) altered the GABA response: the GABA-induced ICl decreased progressively as VH became more negative. 4. The effects of changing the external and internal Na+ concentrations ([Na+]o and [Na+]i) on the GABA-induced ICl were also studied. Increasing [Na+]o at a constant [Na+]i reduced this current while increasing [Na+]i at a fixed [Na+]o facilitated it. 5. A high temperature coefficient of about 3 was estimated with respect to the percentage reduction in GABA-induced ICl due to [Na+]o. 6. These results indicate that the [Na+]o-dependent suppression of GABA-induced ICl was mediated chiefly by the uptake of GABA subserved by a Na-GABA co-transport mechanism. 7. GABA dose-response measurements were made with and without external Na+. The [Na+]o-induced suppression was more pronounced in relative amount at lower concentrations and in absolute amount at intermediate concentrations. Analysis of these data indicates, however, that the Na+-coupled GABA influx kept increasing at GABA concentrations high enough to nearly saturate GABA-induced ICl, and the same saturating level was observed as in the Na+-free case. This indicates that the electrogenic co-transport current was much smaller so that our measurements of GABA-induced ICl' were contaminated very little. Thus, the present method based on recording of GABA-induced ICl was legitimate for the analysis of the Na-GABA co-transport. 8. By analysing the [Na+]o-dependent suppression of GABA-induced ICl, the stoichiometric ratio of the underlying co-transport was estimated to be one: one Na+ ion per GABA molecule. 9. The ICl induced by GABA agonists such as beta-alanine, taurine, l-GABOB (l-gamma-amino-beta-hydroxybutyric acid) and muscimol was not affected by the amount of external Na+ present, suggesting difference in the affinity between receptor and transport carrier.