Representative wild type larva in situ hybridization with cacna1aa antisense and sense probes. Dpf, days post-fertilization; MeO, medulla oblongata; TeO, optic tectum

a Representative western blot of 4 dpf cacna1aa and Ctrl-MO larvae (left panel) and quantification of all samples (right panel). b Dorsal and side views of representative 4 dpf Ctrl-MO and cacna1aa MOs larvae

Locomotor activity of 4 dpf cacna1aa and Ctrl-MO larvae. Larvae were habituated to the apparatus 20 min (10 min light, 10 min in dark) before experiment, and total locomotor activity was tracked within 10 min. The data were pooled from 2 independent experiments. The results were analyzed using one-way ANOVA, followed by Tukey’s post-hoc test. Dots represent individual measurements, the central horizontal mark is the mean, and error bars represent standard deviation (SD) (n = 44–48/group). ^*P < 0.05 vs Ctrl-MO group in relevant phase, ^#P < 0.05 vs Ctrl-MO group in light phase. Dpf, days post-fertilization

Representative electroencephalographic recording illustrating the epileptiform-like discharges recorded in zebrafish Ctrl-MO larvae and cacna1aa morphants (2.5 ng + 2.5 ng). The EEG recordings were obtained from zebrafish larval optic tectum at 4 dpf. a Five-minute-long fragment of representative recording from Ctrl-MO larvae. b Five-minute lasting continuous recording from cacna1aa morphant demonstrating the background and ictal activity. Small letters (c–f) correspond to respective ictal events depicted on traces b–e. c–f Epileptiform-like discharges, high-voltage spikes, spike-wave complexes, and polyspike-wave discharges

Effect of ASDs on epileptiform-like discharges recorded from the optic tectum of 4 dpf cacna1aa and Ctrl-MO morphants. Larvae were incubated (2 h) with different ASDs. Results are presented as a number of events, b mean duration of events (msec), and c cumulative duration of events (msec) during 20 min of recording. Statistical analysis was performed using one-way ANOVA with Tukey’s post-hoc test. Dots represent individual measurements, the central horizontal mark is the mean, and error bars represent SD (n = 9–25). Symbols represent following comparisons: *P < 0.05 vs Ctrl-MO group incubated with Veh, ^#P < 0.05 vs cacna1aa MOs group incubated with Veh. CBZ, carbamazepine (100 μM); ETX, ethosuximide (10 mM), LTG, lamotrigine (200 μM), TPR, topiramate (100 μM), VPA, sodium valproate (100 μM)