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. 2021 Jul 19;11:14733. doi: 10.1038/s41598-021-94282-6

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Stimulation of presynaptic neurons recapitulates spontaneous activity. (a) 50 spontaneous co-occurring spikes in D3 and E5 were used as reference points to identify coupled spikes in A7 (spikes depicted as raster’s below). Spiking activity in A7 after stimulation of electrode D3 using a 3µA biphasic current injection, 200 µsec total duration, 500 times. A blanking period (1.5 ms) during which no voltage data is collected is applied to all electrodes due to artifacts introduced by stimulation. For the stimulation experiments we therefore measured latency from the start of the stimulation period. (b) The spike amplitude distribution of spikes detected at A7 (n = 200 randomly sampled spikes) after spontaneous propagation signals at D3/E5 (n = 572) is not significantly different than the spike distribution of spikes detected at A7 (n = 200 randomly sampled spikes, P = 0.2, two-sample KS test, two-sided, 1 MEA with 2 recording sessions for (a,b)) after stimulation at electrode D3. (c) Correlation of coupling probabilities for spontaneous activity versus stimulated activity. Each data point in (c) and (b) represents coupled neurons. Couplings were identified by identifying spontaneous propagation signal activity as references in a CCG and a coupling probability was assigned to the postsynaptic unit. Following identification of coupling relationships, electrodes associated with presynaptic propagation signals were then stimulated in order to obtain coupling probabilities with the same postsynaptic unit (n = 20 couplings, P = 0.45 paired t-test). (d) For the same coupling events in (c), the correlation between spike amplitudes of the postsynaptic response in the spontaneous condition was compared to the spike amplitudes of the stimulated condition. Error bars represent the standard deviation of spike amplitude distributions (n = 20 couplings, P = 0.18 paired t-test, 3 MEAs and 6 total recording sessions were used for (c,d)).