Fig. 3: Investigation of single-cell electrophysiological properties and somatic VGSC densities in control-, patient-derived and KO neuronal cultures.

Scheme of whole-cell current-clamp recording in 10 weeks old 2D cell culture (a) and representative recording traces depicting evoked neuronal firing (b). c Representative APs recorded from a control and a DCHS1 neuron. Quantification of the AP threshold (d), AP overshoot (e) and AP half-width (f) for control- and patient-derived neurons. g Micrographs of 10 weeks old control- and patient-derived 2D neurons immunostained for NEUN and SCN3A. h Quantification of the somatic SCN3A intensity of control- and patient-derived neurons. i Micrographs of 9 months old control- and patient-derived 3D neurons immunostained for NEUN and SCN3A. j Quantification of the somatic SCN3A intensity of control- and patient-derived 3D neurons. k Scheme of silicon probe recording in a DCHS1 hCO combined with lamotrigine treatment. l Representative traces of neuronal spike activity recorded in a DCHS1 hCO in the absence and presence of lamotrigine. Recordings were performed for 5 min. Quantification of the percentage of spiking rate (m) and burst rate (n) in the absence and presence of lamotrigine. Scale bars: 10 µm (g) and 20 µm (i). Data are represented as mean ± SEM. Statistical significance was based on one-way ANOVA with Turkey’s multiple comparison tests (d–f, h, j) and two-sided unpaired ttest (m, n) (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). Independent wells (d–f, h) or hCOs (j, m, n) were analyzed. Every dot in the plots refers to independently analyzed neurons (d–f, h, j) or independently analyzed recording areas (m, n). At least eighteen (n = 18) randomly chosen neurons or five (n = 5) recording areas were analyzed across three independent batches (N = 3). Source data are provided as a Source Data file, including the exact p-values and n numbers. Created in BioRender. Di Matteo, F. (2025) https://BioRender.com/m95d251, https://BioRender.com/e46e404.