Supplementary Figure 9: Optogenetic characterization of OXT mutants.
From: Zebrafish oxytocin neurons drive nocifensive behavior via brainstem premotor targets
(a) Data from a single clutch of incrossed heterozygotes, stimulated with blue light and monitored behaviorally as described in the main text. Gray = WT, Blue = het, Red = null mutant. Response probability: F(1,168) = 1.14, p=0.29 (homozygous WT vs het), F(1,102) = 3.9, p=0.051 (WT vs null), F(1,126) = 1.99, p= 0.16 (het vs null); Response frequency: F(1,168) = 0.48, p=0.49 (WT vs het), F(1,102) = 5.71, *p=0.017 (WT vs null), F(1,126) = 3.55, p = 0.062 (het vs null); Number of bouts: F(1,168) = 0.28, p=0.59 (WT vs het), F(1,102) = 6.15, *p = 0.015 (WT vs null), F(1,126) = 4.52, *p=0.036 (het vs null); n = 13 (WT), 17 (het), 6 (null), two-way ANOVA. Center black line = Mean, Box = 1 SD, range = 95% confidence interval. (b) Data from a single clutch (het crossed to null mutant). Response probability: F(1,210) = 8.29, **p=0.0044 (het vs null); Response frequency: F(1,210) = 12.46, ***p = 0.0005 (het vs null); Number of bouts: F(1,210) = 13.66, ***p= 0.0003 (het vs null); n = 21 (het), 16 (null), two-way ANOVA. Center black line = Mean, Box = 1 SD, range = 95% confidence interval. (c) Comparing ChR2-induced responses in heterozygous and WT fish (het crossed to WT, 2 clutches pooled). Response probability: F(1,185) = 0.02, p=0.90 (WT vs het); Response frequency: F(1,185) = 0.62, p=0.43 (WT vs het); Number of bouts: F(1,185) = 0.91, p=0.34 (WT vs het); n = 10 (WT), 21 (het), two-way ANOVA. Center black line = Mean, Box = 1 SD, range = 95% confidence interval. (d) While the absolute number of ChR2-evoked tail bends was smaller in null mutants than in heterozygous siblings, we found no significant differences in the mean peak turn angles between heterozygous and null mutants for a 5s stimulation duration. Mean peak tail angles: 75.54 ± 2.16° (hets) vs 68.18 ± 1.28° (null), n =704 (het), 253 (null) detected bouts, p = 0.53, two-sided Wilcoxon rank-sum test.
