Neuron-glia crosstalk revealed in reverberating networks by simultaneous extracellular recording of spikes and astrocytes' glutamate transporter and K+ currents

Astrocytes reuptake synaptically released glutamate with electrogenic transporters (GluT), and buffer the spike-dependent extracellular K(+) ([K(+)]o) excess with background K(+) channels. We studied neuronal spikes and the slower astrocytic signals on reverberating neocortical cultures and organotypic slices from mouse brains. Spike trains and glial responses were simultaneously captured from individual sites of multi-electrode arrays (MEA), by splitting the recorded traces into appropriate filters, and reconstructing the original signal by deconvolution. GluT currents were identified by using DL-threo-beta-benzyloxyaspartate (TBOA). K(+) currents were blocked by 30 µM Ba(2+), suggesting a major contribution of inwardly rectifying K(+) currents (Kir) Both types of current were tightly correlated with the spike rate, and their astrocytic origin was tested in primary cultures by blocking glial proliferation with cytosine β-d-arabinofuranoside (AraC). The spike-related, time-locked inward and outward K(+) currents in different regions of the astrocyte syncytium were consistent with the assumptions of the spatial K(+) buffering model. In organotypic slices from ventral tegmental area (VTA) and prefrontal cortex (PFC), the GluT current amplitudes exceeded those observed in primary cultures by several orders of magnitude, which allowed to directly measure transporter currents with a single electrode. Simultaneously measuring cell signals displaying widely different amplitudes and kinetics will help clarify the neuron-glia interplay, and make it possible to follow the cross-talk between different cell types in excitable as well as non-excitable tissue.