Mechanisms of GABA release from human astrocytes

• Published in: Glia Vol. 59; no. 11; pp. 1600 - 1611
• Main Authors: Lee, Moonhee, McGeer, Edith G., McGeer, Patrick L.
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.11.2011
• Subjects: 4-Aminobutyrate transaminase; Astrocytes; Astrocytes - chemistry; Astrocytes - metabolism; Blotting, Western; Calcium; Calcium (intracellular); Calcium - analysis; Calcium - metabolism; calcium chelation; Cell culture; Cell Survival - drug effects; Cells, Cultured; Chelating agents; Chelating Agents - pharmacology; Culture Media; Cyclohexanecarboxylic Acids - pharmacology; D-serine; Data processing; DNA Primers; Egtazic Acid - analogs & derivatives; Egtazic Acid - pharmacology; Enzymes; Excitatory Amino Acid Agonists - pharmacology; GABA receptors; GABA transporters; gamma -Aminobutyric acid A receptors; gamma -Aminobutyric acid B receptors; gamma -Aminobutyric acid transporter; gamma-Aminobutyric Acid - metabolism; glutamate; Glutamate decarboxylase; glutamate receptors; Glutamic acid; Glutamic Acid - pharmacology; Glutamic acid receptors; Glutamic acid receptors (ionotropic); Glycine; Humans; mRNA; N-Methyl-D-aspartic acid receptors; Neurotransmitters; Real-Time Polymerase Chain Reaction; Receptors, Glutamate - metabolism; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; Vesicular Glutamate Transport Proteins - antagonists & inhibitors; Vesicular Glutamate Transport Proteins - genetics; Vesicular Glutamate Transport Proteins - metabolism; Vigabatrin - pharmacology
• ISSN: 0894-1491; 1098-1136; 1098-1136
• DOI: 10.1002/glia.21202

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 GABAA and GABAB 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 Ca2+‐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 Ca2+ concentration ([Ca2+]i) and was reduced by adding the Ca2+ 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. © 2011 Wiley‐Liss, Inc.