title: Mechanisms of GABA release from human astrocytes.
abstract: We have previously demonstrated that human astrocytes are GABAergic cells. Throughout the adult human brain, they express the GABA synthesizing enzyme GAD 67, the GABA metabolizing enzyme GABA-T, and the GABA(A) and GABA(B) receptors. GABA modulates the actions of microglia, indicating an important role for astrocytes beyond that of influencing neurotransmitter function. Here we report on the mechanisms by which astrocytes release GABA. Astrocytes were found to express the mRNA and protein for multiple GABA transporters, and multiple receptors for glutamate, GABA, and glycine. In culture, untreated human astrocytes maintained an intracellular GABA level of 2.32 mM. They exported GABA into the culture medium so that an intracellular-extracellular gradient of 3.64 fold was reached. Inhibitors of the GABA transporters GAT1, GAT2, and GAT3, significantly reduced this export in a Ca(2+)-independent fashion. Intracellular GABA levels were enhanced by treatment with the GABA-T inhibitors gabaculine or vigabatrin. Treatment with glutamate increased GABA release in a concentration-dependent fashion. This was partially inhibited by blockers of N-methyl-D-aspartate and kainate receptors. Conversely, glycine and D-serine, co-agonists of NMDA receptors, enhanced the GABA release. GABA release was accompanied by an increase in intracellular Ca(2+) concentration ([Ca(2+)](i)) and was reduced by adding the Ca(2+) chelator, BAPTA-AM to the medium. These data indicate that astrocytes continuously synthesize GABA and that there are multiple mechanisms which can mediate its release. Each of these may play a role in the physiological functioning of astrocytes.
authors: Kinsmen Laboratory of Neurological Research, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
authors: Moonhee
authors: M
authors: Lee
authors: Edith G
authors: EG
authors: McGeer
authors: Patrick L
authors: PL
authors: McGeer
mesh_terms: D001253:Astrocytes / Q000737:chemistry / Q000378:metabolism*; D015153:Blotting, Western; D002118:Calcium / Q000032:analysis / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D003470:Culture Media; D003509:Cyclohexanecarboxylic Acids / Q000494:pharmacology; D017931:DNA Primers; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D018698:Glutamic Acid / Q000494:pharmacology; D006801:Humans; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D017470:Receptors, Glutamate / Q000378:metabolism; D050496:Vesicular Glutamate Transport Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D020888:Vigabatrin / Q000494:pharmacology; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
chemical_list: D002614:Chelating Agents; D003470:Culture Media; D003509:Cyclohexanecarboxylic Acids; D017931:DNA Primers; D018690:Excitatory Amino Acid Agonists; D012333:RNA, Messenger; D017470:Receptors, Glutamate; D050496:Vesicular Glutamate Transport Proteins; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; C012987:gabaculine; D018698:Glutamic Acid; D004533:Egtazic Acid; D005680:gamma-Aminobutyric Acid; D020888:Vigabatrin; D002118:Calcium