Fig. 4

a Methodological workflow of behavioral assays in TL and panx1 knockout larvae. b Larvae’s (n = 36/genotype) baseline and 15 mM PTZ-induced activity (Δpixel ± s.e.m.) were scored. Panx1b^−/− baseline activity was higher than TL (P = < 1.0 × 10⁻⁴). PTZ induced hyperactivity peaked at 15–30 min for TL (gray), was reduced in DKO (light blue), significantly reduced in panx1a^−/− (dark blue, P = < 1.0 × 10⁻⁴) for the last 40 min, and significantly increased in panx1b^−/− larvae (magenta, P = 1.3 × 10⁻³) around 20 min compared to TL; Two-way repeated measures ANOVA and Bonferroni multiple comparisons. Dashed lines indicate TL’s max average activity score. c AUC derived from the activity plots in (b) significantly differed between TL and panx1b^−/− baselines (open points; P < = 1.0 × 10⁴). AUC for 1 h post-PTZ treatment was significantly reduced for panx1a^−/− (P = < 1.0 × 10⁻⁴) and increased for panx1b^−/− (filled points; P = 5.7 × 10⁻³). Change in PTZ treatment from average baseline activity was significantly reduced in panx1a^−/− compared to TL (P = 2.0 × 10⁻⁴; Tukey’s multiple comparisons test). d Stage II count (rapid circling) did not differ among groups (n = 18; count/2 min). Stage III (convulsion) was significantly reduced in DKO (P = 1.5 × 10⁻²) and panx1a^−/− (P = 0.02; Two-way repeated measures ANOVA) compared to TL. Dashed lines represent max average stage II and stage III counts for TL. e Time course of average stage III counts differed between the two groups, peaking at 26 min post-PTZ treatment for TL and 34 min for panx1b^−/−. Time course where >50% of the larvae reached 1–5 stage III counts revealed a delayed peak onset for panx1b^−/−. f IEG upregulation after 1 h PTZ treatment, represented as fold-change for PTZ treated larvae against non-treated controls. Panx1 knockout larvae had reduced upregulation of the IEGs compared to TL, with DKO and panx1b^−/− showing the greatest reduction. (N = 3 experimental replicates). g Survival rate of larvae (n = 80) post-PTZ treatment was significantly higher for panx1a^−/− followed by panx1b^−/−, and the survival rate of DKOs did not differ from TL. Survival at 24 h: TL: 1.25%, DKO: 11%; P = 0.38, panx1a^−/−: 30%; P = 8.8  × 10⁻³, panx1b^−/−: 13%; P = 8.1 × 10⁻³; Mantel–Cox test. h SOM revealed similarities and differences in PTZ-induced activity and bursting behaviors of TL and panx1 knockouts (n = 36). TL and DKO were most similar, panx1a^−/− differed the most from TL (P < 0.0001; Fisher’s exact test), and panx1a^−/− and panx1b^−/− were most distinct (P < 0.0001). Values and color represent the degree of similarity. N = number of larvae. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.