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 (?pixel?±?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.