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GABA (gamma-aminobutyric acid) is a transmitter with dual action. Whereas it excites neurons during the first week of postnatal development, it represents the major inhibitory transmitter in the mature brain. GABA also activates astrocytes by binding to ionotropic (GABA(A)) and metabotropic (GABA(B)) receptors. This results in glial calcium transients which can induce the release of gliotransmitters, rendering GABA an important mediator of neuron-glia interaction. Using whole-cell patch-clamp and ratiometric calcium imaging in hippocampal slices from rats at postnatal days 3-34, we have analyzed the developmental profile as well as the cellular mechanisms of calcium signals induced by GABA(A) and GABA(B) receptor activation in astrocytes. We found that GABA-evoked glial calcium transients are mediated by both GABA(A) and GABA(B) receptors. Throughout development, GABA(A)-receptor activation resulted in immediate calcium transients in the vast majority of astrocytes, most likely by influx of calcium through voltage-gated calcium channels. GABA(B) receptor activation, in contrast, resulted in delayed calcium transients, which were blocked following depletion of intracellular calcium stores and during persistent activation of heterotrimeric G-proteins. GABA(B) receptor-mediated calcium signals exhibited a clear developmental profile with less than 10% of astrocytes responding at P3 or P32-34, and about 60% of cells between P11 and P15. Our data thus indicate that GABA(B) receptor-mediated calcium transients are due to calcium release from intracellular stores following G-protein activation. Moreover, GABA(B) receptor-mediated calcium signaling in astrocytes preferentially occurs at a period during postnatal development when hippocampal networks are established.

Glial cells express specific high-affinity transporters for glutamate that play a central role in glutamate clearance at excitatory synapses in the brain. These transporters are electrogenic and are mainly energized by the electrochemical gradient for sodium. In the present study, we combined somatic whole-cell patch-clamp recordings with quantitative Na+ imaging in fine cellular branches of cerebellar Bergmann glial cells and in dendrites of Purkinje neurons to analyze intracellular Na+ signals close to activated synapses. We demonstrate that pressure application of glutamate and glutamate agonists causes local Na+ signals in the mM range. Furthermore, we analyzed the pharmacological profile, as well as the time course and spatial distribution of Na+ signals following short synaptic burst stimulation of parallel or climbing fibers. While parallel fibers stimulation resulted in local sodium transients that were largest in processes close to the stimulation pipette, climbing fibers stimulation elicited global sodium transients throughout the entire cell. Glial sodium signals amounted to several mM, were mainly caused by sodium influx following inward transport of glutamate and persisted for tens of seconds. Sodium transients in dendrites of Purkinje neurons, in contrast, were mainly caused by activation of AMPA receptors and had much faster kinetics. By reducing the driving force for sodium-dependent glutamate uptake, intracellular sodium accumulation in glial cells upon repetitive activity might provide a negative feedback mechanism, promoting the diffusion of glutamate and the activation of extrasynaptic glutamate receptors at active synapses in the cerebellum.