Short-latency cross- and autocorrelation identify clusters of interacting cortical neurons recorded from multi-electrode array

Spontaneous bursting activity is present in vivo during CNS development and in vitro in neocortex slices. A prerequisite for understanding the cooperative behavior in neuronal ensembles is large-scale simultaneous extracellular electrophysiology by using either “tetrodes” (4-wire electrode) in awake animals or multi-electrode arrays (MEA) in long-term cultured networks as we did here. We show that from a single low-noise MEA electrode it is possible to identify up to 3–4 types of waveforms whose timestamps show excitatory and inhibitory short-latency (2–4 ms) cross-correlations, indicative of monosynaptic connections. Moreover, the MEA units autocorrelagrams (AC) resulted to have behaviors similar to those demonstrated in vivo by using tetrodes or shanks. Principal component analysis of AC followed by a K-means classification returned 3–4 different clusters whose firing- and burst-related properties were typical of assemblies of putative excitatory and inhibitory neurons. By manipulating the networks with a GABAA antagonist (gabazine), we could detect cell groups selectively responding to blockade of GABA transmission with IC50s of 82 ± 2 and 770 ± 70 nM. These methods, expanded to organotypic co-cultures of CNS regions may be useful to better understand their connecting properties in studies of regenerative medicine.