GABA- and glycine-mediated fall of intracellular pH in rat medullary neurons in situ.
In the region of the ventral respiratory group in brain stem slices from neonatal rats, intracellular pH (pHi) and membrane currents (I(m)) or potentials were measured in neurons dialyzed with the pH-sensitive dye 2',7'-bis-carboxyethyl-5(6)-carboxyfluorescein. Currents and increases in membrane conductance (g(m)) during bath application of 0.1 or 1 mM gamma-aminobutyric acid (GABA) were accompanied by a delayed mean fall of pHi by 0.17 and 0.25 pH units, respectively, from a pHi baseline of 7.33. These effects were reversibly suppressed by 50-100 microM bicuculline. Similar effects on I(m), g(m), and pHi were revealed on administration of 0.1 or 1 mM glycine. These responses were abolished by 10-100 microM strychnine. Dialysis of the cells with 15-30 microM carbonic anhydrase led to an acceleration of the kinetics and a potentiation of the GABA-induced pHi decrease. GABA- and glycine-evoked pHi decreases were very similar during recordings with either high- or low-Cl- patch electrodes, although the reversal potential of the accompanying currents differed by approximately 60 mV. The GABA-induced pHi decrease, but not the accompanying I(m) and g(m) responses, was suppressed in CO2/HCO3(-)-free, N-2-hydroxy-ethylpiperazine-N'-2-ethane sulphonic acid pH-buffered solution. Depolarization from -60 to +30 mV resulted in a sustained fall of pHi by maximally 0.5 pH units. In this situation, the GABA-induced fall of pHi turned into an intracellular alkalosis of 0.09-0.15 pH units. The results confirm and extend previous findings obtained in vivo that GABA- or glycine-induced intracellular acidosis of respiratory neurons is due to efflux of HCO3- via the receptor-coupled Cl- channel.
Lückermann
M
M
II. Physiologisches Institut, Universität Göttingen, Germany.
Trapp
S
S
Ballanyi
K
K
D000472:Alkalosis, Respiratory / Q000503:physiopathology; D000818:Animals; D001639:Bicarbonates / Q000494:pharmacology; D002245:Carbon Dioxide / Q000494:pharmacology; D002255:Carbonic Acid / Q000494:pharmacology; D018118:Chloride Channels / Q000187:drug effects / Q000502:physiology; D005260:Female; D005998:Glycine / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D008297:Male; D008526:Medulla Oblongata / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012119:Respiration / Q000187:drug effects / Q000502:physiology*; D005680:gamma-Aminobutyric Acid / Q000502:physiology*
D001639:Bicarbonates; D018118:Chloride Channels; D000644:Quaternary Ammonium Compounds; D002245:Carbon Dioxide; D002255:Carbonic Acid; D005680:gamma-Aminobutyric Acid; D005998:Glycine