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. 2018 Jul 19;8:10957. doi: 10.1038/s41598-018-29069-3

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© The Author(s) 2018

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Acute L-glutamate treatment impaired network activity and synchrony immediately and over a week post-administration. (A) Experimental schedule for ESC-derived neuron seeding, culture and L-glutamate treatment. In the treatment group, 100 µM of L-glutamate was administered at day14 for 20 min then washed out. MEA recordings were performed from week 1 to week 3. (B) Scatter distribution of the mean burst firing rate (mBR; Burst event/min; x-axis) against the mean firing rate (mFR; log of spike/sec; y-axis) for individual unit recorded from CTR (black filled circles) and L-glut (black open circle) groups. Data is shown for week 2 (left panel) and week 3 (right panel). The normalized distribution CTR (black line) and L-glut (gray line) groups are displayed as a projection of each axis. The dashed black line represents the average for the CTR group. The dotted black line represents the average for the L-glut group. mFR: mean firing rate for individual electrodes; mBR: mean bursting rate for individual electrodes. (C) ESC-derived neurons after L-glutamate treatment (L-glut; 100 µM for 20 min at day 14) at week 2 (top panels) and week 3 (bottom panels). From left to right panels are shown SOX-1, βIII-Tubulin and merged image of both fluorescent markers. Scale = 200 μm. (D) Change at week 2 and week 3 (expressed as a Z-score from week1 baseline) of the number of bursting cells (left panel) and weighted mean firing rate (wMFR; right panel). For each group, the number of wells with significantly increasing change over the total number of wells recorded is shown above the scatter plot. wMFR: weighted mean population firing rate. (E) Change in network synchrony at week 2 and week 3 (expressed as a Z-score from week1 baseline) for event synchronization (left panel), cross-correlation peak (middle panel) and mean network burst firing rate (mBFR; right panel). For each group, the number of wells with significantly increasing change over the total number of wells recorded is shown above the scatter plot. For post-hoc two-sample test *, ** and *** indicate p < 0.05, p < 0.01 and p < 0.001, respectively. mBFR: mean population burst firing rate.