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Fig. 3 Sleep induction by optogenetic stimulation of QRFP/Pth4 neurons requires pth4 but not qrfp (A–D) Optogenetic stimulation of QRFP/Pth4 neurons suppresses locomotor activity and increases sleep in qrfp+/− (A and B) and qrfp−/− fish (C and D) compared with non-transgenic siblings. (E) FISH shows that pth4 and qrfp are co-expressed in the same hypothalamic neurons (586/589 cells, n = 23 fish). Scale bar, 25 μm. (F–I) Optogenetic stimulation of QRFP/Pth4 neurons suppresses locomotor activity and increases sleep in pth4+/+ fish (F and G) but not in pth4−/− fish (H and I) compared with non-transgenic siblings. Mean ± SEM line plots and Tukey boxplots are shown. n = number of fish. ∗∗∗∗p < 0.0001, ns, p > 0.05 by Mann-Whitney U test. See also Figures S2 and S3.