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Fig. 2 Two-photon laser ablation of DRN5-HT neurons retarded emergence from propofol anesthesia. A, Behavioral experiments diagram. B, The representative images of two-photon laser ablation on DRN5-HT neurons. C, Dose response curve of propofol on the loss-of-response to tail-fin pinch. N = 21 larvae/concentration. D,E, Specific ablation of DRN5-HT had little effect on the induction time of 30 μM propofol anesthesia detected by mechanical stimulation (D, control group, 247.1 ± 12.1 min; mock ablation, 250.6 ± 10.7 min; DRN ablation, 249.4 ± 11.2 min; p > 0.05; one-way ANOVA) and loss of spontaneous movement (E, control group, 37.4 ± 3.4 min; mock ablation, 34.1 ± 3.6 min; DRN ablation, 36.4 ± 3.2 min; p > 0.05; one-way ANOVA). F,G, Specific ablation of DRN5-HT significantly prolonged emergence time from 30 μM propofol, detected by both recovery to mechanical stimulation (F, F = 5.91, control vs DRN ablation; 11.5 ± 1.0 min vs 16.5 ± 1.7 min; p = 0.019; one-way ANOVA) and the recovery time of spontaneous movement (G, F = 5.70; control vs DRN ablation; 81.0 ± 3.7 min vs 95.4 ± 4.2 min; p = 0.036; one-way ANOVA). H–J, Specific ablation of DRN5-HT had little effect on the induction time of 3 μM propofol anesthesia detected by mechanical stimulation (H, loss of response rate at 30 min; I, induction time, control group, 943.2 ± 53.8 s; mock ablation, 990.9 ± 63.6 s; DRN ablation, 900.0 ± 50.6 s; p > 0.05; one-way ANONA) and loss of spontaneous movement (J, control group, 472.6 ± 52.4 s; mock ablation, 495.3 ± 54.1 s; DRN ablation, 530.3 ± 59.9 s; p > 0.05; one-way ANOVA). K, Specific ablation of DRN5-HT had little effect on the induction time of 1 µM propofol anesthesia detected by loss of spontaneous movement (control group, 776.0 ± 64.5 s; mock ablation, 880.6 ± 51.8 s; DRN ablation, 876.2 ± 50.1 s; p > 0.05; one-way ANOVA). The sample size for each group is indicated in parentheses. In H, the values in parentheses are the ratio of the number of larvae that lost response to the total sample size.