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Locomotor activity of larvae devoid of opn4xa-mediated photosensitivity (opn4xa-/-) in LD, DD and LL. A) Experimental design of LD, DD and LL experiments. White rectangles represent the day period, black rectangles represent the night period, dark grey rectangles represent the subjective day period and light grey rectangles the subjective night. For each experiment, larvae are entrained for 5 LD cycles and their locomotor activity is tracked either in LD (LD), constant darkness (DD) or constant light (LL) the larvae are therefore 5dpf at the beginning of locomotor activity measurements. B) Average distance moved (mm/min over 10min) of 3 independent experiments in LD. Mean ± SE. The distance moved is not different in opn4xa+/+ and opn4xa-/- larvae during the 1st (p = 0.73), 2nd days (p = 0.50) and 3rd days (p = 0.07) nor during the 1st (p = 0.30), and 2nd nights (p = 0.27) (S4 Table). A lower level of activity is found in opn4xa-/- larvae during the 3rd night (p = 0.01) but is visually clear in only one of the 3 independent experiments (Mann-Whitney two-tailed test). C) Average distance moved (mm/min over 10min) of 3 independent experiments in DD. Mean ± SE. D) Estimation of the periods using the FFT-NLLS method calculated over four cycles. The mean period is not significantly different between control and opn4xa+/+ and opn4xa-/- larvae in DD (opn4xa+/+: 25.05 ± 1.43 hours (n = 64), opn4xa-/-: 25.35 ± 1.60 hours (n = 60); mean±SD; p = 0.29; Mann-Whitney two-tailed test). Mean± sd (in hours) is represented. Each grey point represents a larva. E) Average distance moved (mm/min over 10min) of 4 independent experiments in LL. Mean ± SE. opn4xa-/- are more active than controls during the first night (p = 0.02, see S6 Table). F) Estimation of the periods using the FFT-NLLS method calculated over three cycles. The mean period is significantly different between opn4xa+/+ and opn4xa-/- larvae in LL. Mean± sd (in hours) is represented. Each grey point represents a larva.
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