Net uptake of gamma-aminobutyric acid by a high affinity synaptosomal transport system.
Reuptake of gamma-aminobutyric acid (GABA) by a high affinity transport system in nerve endings in the central nervous system is thought to terminate the action of this postulated neurotransmitter. This hypothesis has been challenged since the demonstration of exchange between synaptosomal and exogenous GABA (G. Levi and M. Raiteri, Nature 250: 735, 1974). In our studies, rat cortical synaptosomes were incubated (25 degrees C) in various media containing 10 muM 14C-GABA. After the synaptosomes were removed by centrifugation, 14C and total GABA (fluorometric assay) in the resulting supernatant were measured. Uptake of labeled GABA, detected by a decrease in medium radioactivity, is Na+- and K+-dependent. Net GABA uptake, however, does not parallel 14C-GABA translocation. Exchange accounts for 20 to 70% of radiolabeled GABA accumulation depending upon the experimental conditions. On the other hand, GABA-deficient synaptosomes (prepared by treatment with 56 mM KCl and 1 mM CaCl2) show equivalent net and radiolabeled GABA uptake in Ringer's solution containing 1 to 4 mM KCl and 60 to 150 mM NaCl (average 4.6 nmol of GABA accumulated per mg of synaptosomal protein). Net and 14C-GABA uptake by GABA-deficient synaptosomes are identical at various pH values (6.0-8.5), synaptosomal protein concentrations (0.4-3.5 mg/ml) and temperatures (5-37 degrees C). Although GABA homoexchange may contribute significantly to radiolabel accumulation by synaptosomes containing higher GABA levels (9.5-9.9 nmol/mg), homoexchange is limited in GABA-depleted synaptosomes. Our results are consistent with the proposal that presynaptic GABA capture by a high affinity system in vivo may terminate the action of this neuroactive amino acid.
D000613:Aminobutyrates / Q000378:metabolism*; D000818:Animals; D001693:Biological Transport, Active; D002540:Cerebral Cortex / Q000378:metabolism / Q000648:ultrastructure; D006358:Hot Temperature; D066298:In Vitro Techniques; D008297:Male; D051381:Rats; D012964:Sodium / Q000494:pharmacology; D013574:Synaptosomes / Q000378:metabolism*; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
D000613:Aminobutyrates; D005680:gamma-Aminobutyric Acid; D012964:Sodium