Fig. 7

Global activity in response to dark-light preference test in 14 dpf larvae prenatally exposed. A. Average movement of control larvae across the dark-light preference test protocol. A reduction in movement can be observed during the strobe light phase as at 8 dpf. B. Smoothing of the average response for each condition across the entire experiment, and a linear mixed-model analysis was performed across the different phases independently. During the initial light phase, all conditions showed a log2 fold change below 1, but over that phase, animals in groups E, M, N, EM, and EN became more similar to the control. During the strobe phase, groups N, MN, and EMN showed an increased log2 fold change. Finally, after the strobe light, two divergent effects were observed: E, M, and EM animals became more different from the control over time, while N, EN, and NM groups exhibited the opposite trend. Interestingly, these effects differed between ethanol- and nicotine-exposed groups. C. Most of the exposed animals were hypoactive, especially in the EMN and MN groups during the whole experiment. D. Most of the exposed animals were hypoactive, especially in the EMN group. B: # p < 0.05 during the initial light phase (0–600 s.), $ p < 0.05 during the strobe-light phase (600–780 s.) and * p < 0.05 after the strobe phase (780–1380 s.). For each phase, the difference is compared to control animals. P-values were obtained using a linear mixed-effects model with condition over time as a fixed effect and subject (larva ID) and Batch as random effects. C-D: * = p < 0.05 compared to control animals. P-values were calculated using Firth's bias-reduced logistic regression to compare each treatment to control, followed by Benjamini–Hochberg correction for multiple comparisons. n ≥ 12 per condition.