Neuron-glia cross talk revealed in reverberating networks by simultaneous extracellular recording of spikes and astrocytes' glutamate transporter and K+ currents.
Astrocytes uptake synaptically released glutamate with electrogenic transporters (GluT) and buffer the spike-dependent extracellular K+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 multielectrode 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-β-benzyloxyaspartate (TBOA). K+currents were blocked by 30 μM Ba2+, suggesting a major contribution of inwardly rectifying K+currents. 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 and prefrontal cortex, 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 nonexcitable tissue.
Department of Biotechnologies and Biosciences and Milan Center For Neuroscience (NeuroMI), University of Milano-Bicocca, Milan, Italy; and.
Enzo
E
Wanke
Department of Biotechnologies and Biosciences and Milan Center For Neuroscience (NeuroMI), University of Milano-Bicocca, Milan, Italy; and.
Francesca
F
Gullo
Department of Biotechnologies and Biosciences and Milan Center For Neuroscience (NeuroMI), University of Milano-Bicocca, Milan, Italy; and.
Elena
E
Dossi
Research Centre "E. Piaggio" and Department of Information Engineering, School of Engineering, University of Pisa, Pisa, Italy.
Gaetano
G
Valenza
Department of Biotechnologies and Biosciences and Milan Center For Neuroscience (NeuroMI), University of Milano-Bicocca, Milan, Italy; and andrea.becchetti@unimib.it.
Andrea
A
Becchetti
D000200:Action Potentials / Q000502:physiology*; D027322:Amino Acid Transport System X-AG / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D001224:Aspartic Acid / Q000494:pharmacology; D002450:Cell Communication / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D004558:Electric Stimulation; D005071:Evoked Potentials / Q000187:drug effects / Q000502:physiology; D066298:In Vitro Techniques; D051379:Mice; D009415:Nerve Net / Q000502:physiology*; D009457:Neuroglia / Q000187:drug effects / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D018377:Neurotransmitter Agents / Q000494:pharmacology; D009924:Organ Culture Techniques; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism*; D043324:beta-Fructofuranosidase / Q000494:pharmacology
D027322:Amino Acid Transport System X-AG; D018377:Neurotransmitter Agents; C120673:benzyloxyaspartate; D001224:Aspartic Acid; D043324:beta-Fructofuranosidase; D011188:Potassium
AraC; K+ buffering; MEA; TBOA; neocortex
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