title: GABA uptake-dependent Ca(2+) signaling in developing olfactory bulb astrocytes.
abstract: We studied GABAergic signaling in astrocytes of olfactory bulb slices using confocal Ca(2+) imaging and two-photon Na(+) imaging. GABA evoked Ca(2+) transients in astrocytes that persisted in the presence of GABA(A) and GABA(B) receptor antagonists, but were suppressed by inhibition of GABA uptake by SNAP 5114. Withdrawal of external Ca(2+) blocked GABA-induced Ca(2+) transients, and depletion of Ca(2+) stores with cyclopiazonic acid reduced Ca(2+) transients by approximately 90%. This indicates that the Ca(2+) transients depend on external Ca(2+), but are mainly mediated by intracellular Ca(2+) release, conforming with Ca(2+)-induced Ca(2+) release. Inhibition of ryanodine receptors did not affect GABA-induced Ca(2+) transients, whereas the InsP(3) receptor blocker 2-APB inhibited the Ca(2+) transients. GABA also induced Na(+) increases in astrocytes, potentially reducing Na(+)/Ca(2+) exchange. To test whether reduction of Na(+)/Ca(2+) exchange induces Ca(2+) signaling, we inhibited Na(+)/Ca(2+) exchange with KB-R7943, which mimicked GABA-induced Ca(2+) transients. Endogenous GABA release from neurons, activated by stimulation of afferent axons or NMDA application, also triggered Ca(2+) transients in astrocytes. The significance of GABAergic Ca(2+) signaling in astrocytes for control of blood flow is demonstrated by SNAP 5114-sensitive constriction of blood vessels accompanying GABA uptake. The results suggest that GABAergic signaling is composed of GABA uptake-mediated Na(+) rises that reduce Na(+)/Ca(2+) exchange, thereby leading to a Ca(2+) increase sufficient to trigger Ca(2+)-induced Ca(2+) release via InsP(3) receptors. Hence, GABA transporters not only remove GABA from the extracellular space, but may also contribute to intracellular signaling and astrocyte function, such as control of blood flow.
authors.affiliation: Abteilung für Allgemeine Zoologie, Universität Kaiserslautern, Postfach 3049, D-67653 Kaiserslautern, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Doengi
authors.forename: Daniela
authors.initials: D
authors.lastname: Hirnet
authors.forename: Philippe
authors.initials: P
authors.lastname: Coulon
authors.forename: Hans-Christian
authors.initials: HC
authors.lastname: Pape
authors.forename: Joachim W
authors.initials: JW
authors.lastname: Deitmer
authors.forename: Christian
authors.initials: C
authors.lastname: Lohr
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000502:physiology*; D002118:Calcium / Q000502:physiology*; D058787:GABA-A Receptor Antagonists; D051379:Mice; D009474:Neurons / Q000187:drug effects / Q000502:physiology; D009830:Olfactory Bulb / Q000187:drug effects / Q000502:physiology*; D011724:Pyridazines / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D005680:gamma-Aminobutyric Acid / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D058787:GABA-A Receptor Antagonists; D011724:Pyridazines; D005680:gamma-Aminobutyric Acid; C049853:gabazine; D002118:Calcium