title: Metabolism of [1,6-(13)C]glucose and [U-(13)C]glutamine and depolarization induced GABA release in superfused mouse cerebral cortical mini-slices.
abstract: Mouse cerebral cortical mini-slices were used in a superfusion system to monitor depolarization-induced (55 mM K(+)) release of preloaded [2,3-(3)H]GABA and to investigate the biosynthesis of glutamate, GABA and aspartate during physiological and depolarizing (55 mM K(+)) conditions from either [1,6-(13)C]glucose or [U-(13)C]glutamine. Depolarization-induced GABA release could be reduced (50%) by the GABA transport inhibitor tiagabine (25 microM) or by replacing Ca(2+) with Co(2+). In the presence of both tiagabine and Co(2+) (1 mM), release was abolished completely. The release observed in the presence of 25 microM tiagabine thus represents vesicular release. Superfusion in the presence of [1,6-(13)C]glucose led to considerable labeling in the three amino acids, the labeling in glutamate and aspartate being increased after depolarization. This condition had no effect on GABA labeling. For all three amino acids, the distribution of label in the different carbon atoms revealed on increased tricarboxylic acid (TCA) activity during depolarization. When [U-(13)C]glutamine was used as substrate, labeling in glutamate was higher than that in GABA and aspartate and the fraction of glutamate and aspartate being synthesized by participation of the TCA cycle was increased by depolarization, an effect not seen for GABA. However, GABA synthesis reflected TCA cycle involvement to a much higher extent than for glutamate and aspartate. The results show that this preparation of brain tissue with intact cellular networks is well suited to study metabolism and release of neurotransmitter amino acids under conditions mimicking neural activity.
authors.affiliation: Department of Pharmacology and Pharmacotherapy, Faculty of Pharmaceutical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark.
authors.forename: Helle S
authors.initials: HS
authors.lastname: Waagepetersen
authors.forename: Søren
authors.initials: S
authors.lastname: Døring
authors.forename: Arne
authors.initials: A
authors.lastname: Schousboe
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002247:Carbon Isotopes / Q000737:chemistry*; D002540:Cerebral Cortex / Q000378:metabolism*; D003035:Cobalt / Q000378:metabolism; D018755:GABA Agonists / Q000378:metabolism; D005947:Glucose / Q000031:analogs & derivatives / Q000378:metabolism; D005973:Glutamine / Q000737:chemistry / Q000378:metabolism; D006706:Homeostasis; D051379:Mice; D009557:Nipecotic Acids / Q000378:metabolism; D000078308:Tiagabine; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D002247:Carbon Isotopes; D018755:GABA Agonists; D009557:Nipecotic Acids; D005973:Glutamine; D003035:Cobalt; D005680:gamma-Aminobutyric Acid; D005947:Glucose; D000078308:Tiagabine