Fig. 5: NitroSynapsin normalizes spontaneous calcium transients and neural network activity in MHS hiPSC-derived cerebrocortical neurons in 2D cultures.
A Spontaneous neuronal calcium transients recorded from individual Ctrl1 and MHS hiPSC-neurons loaded with Fluo-4 AM. B Quantification of Ca2+ transient frequency for events with rise times <200 ms (individual Ctrl1 and MHS hiPSC-neurons responses in upper panel, and grouped responses in lower panel). C Representative calcium traces showing decrease in spontaneous calcium transient frequency after application of 10 µM NitroSynapsin. D Quantification of calcium transient frequency before and after application of NitroSynapsin. E Quantification of difference in normalized fluorescence (ΔF/F0Drug–ΔF/F0Control) as area under the curve (AUC) in response to NitroSynapsin. F Representative heat maps and raster plots of MEA recordings from Ctrl and MHS hiPSC neurons before (w/o) and after treatment with 5 µM NitroSynapsin. Boxes outline examples of network bursts. G–J Quantification of MEA recordings by mean firing rate, electrode burst frequency (representing bursting of individual neurons), network burst frequency (representing bursting of the entire neural network), and synchronous firing. Data are mean ± SEM. Sample size listed above bars represents number of cells (n) analyzed in 5–10 independent experiments. In D, E, G, H, I and J, responses of each MHS patient’s 2D neurons vs. each control shown on left, with grouped MHS patient vs. controls shown on right. *,#,†p < 0.05, **,##,††p < 0.01, ***, ###, †††p < 0.001, ****,####, ††††p < 0.0001 by ANOVA by Sidak’s post-hoc test for comparison to Ctrl1 (*) or to Ctrl2 (#), or within a group (†) between NitroSynapsin treatment vs. without (w/o) treatment; for panel D, comparison was made by non-parametric Kruskal–Wallis test (see Methods).
