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Fig. 1 Expression and stimulation of ChR2 and bPAC in zebrafish motor neurons alter locomotion behavior (A) Confocal images of Tg[mnx1:Gal4]/Tg[UAS:ChR2-P2A-dTomato] and Tg[mnx1En:bPAC-egfp] transgenic embryos (24 hpf) and of Tg[mnx1:Gal4]/Tg[UAS:ChR2-P2A-dTomato] and Tg[mnx1:Gal4]/Tg[UAS:bPAC-V2A-mCherry] double transgenic larvae (4 dpf). Asterisks mark motor neuron cell bodies, arrowheads motor axons. (B) Spontaneous (dark) and evoked (light) tail movements (STMs/ETMs) in 24 hpf embryos. (C) Head-tail angles in immobilized larvae under dark and blue-light conditions. The maximum head-tail angles were determined for wild-type (WT), bPAC (Tg[mnx1: Gal4]/Tg[UAS: bPAC-V2A-mCherry]), and ChR2 (Tg[mnx1:Gal4]/Tg[UAS:ChR2-P2A-dTomato] transgenic larvae (4 dpf). (D) Bending angles of freely swimming larvae under dark and photostimulated conditions. The bending angles were determined for animals as in (C). (E) Swimming speed was measured for individuals 4 dpf before, during, and after blue light stimulation. (F) Average swimming speed with and without blue light stimulation. (G) Distance traveled during the photostimulation phase. In (B), median (cross), 25/75 quartiles, and min to max are shown; in (C) and (E), means ± SEM; in (D) and (F and G), median and 25/75 quartiles (thick and thin black lines), min to max are shown. in (B), Kruskal-Wallis test with Dunn post hoc test.; in (C?F), two-way ANOVA; in (G), one-way ANOVA, each with Tukey multiple comparisons of means. Statistical significance given as ????p < 0.0001, ???p < 0.001, ??p < 0.01, ?p < 0.05.