Extended Data Fig. 6: Behavioral characterization of opioid responsiveness of Ptchd1 KO mice.

a, Evaluation of activity levels by tracking distance traveled in an open field. Data are from 7 WT and 6 Ptchd1 KO mice. b, Evaluation of activity levels by calculating total distance traveled in an open field over 120 minutes (from panel a). Data are from 7 WT and 6 Ptchd1 KO mice. c, Evaluation of activity levels by tracking distance traveled in an open field in first 10 minutes and last 10 minutes from panel a. Data are from 7 WT and 6 Ptchd1 KO mice. d, Evaluation of activity levels by tracking distance traveled in an open field after naltrexone or saline injection. Data are from 7 WT and 6 Ptchd1 KO mice. e, Analysis of time course of morphine analgesia by hot plate assay upon systemic morphine administration. Data are from 7 WT and 6 Ptchd1 KO mice. f, Evaluation of morphine induced analgesia in hot plate assay upon intracerebroventricular (ICV) injection of morphine. Data are from 7 male mice per genotype. g, Analysis of time course of morphine analgesia in the hot plate assay following ICV injection of morphine. Data are from 7 male mice per genotype. h, Evaluation of analgesia by hot plate assay with systemic fentanyl administration. Data are from 7 male mice per genotype. i, Analysis of time course of fentanyl analgesia by hot plate assay from the experiment in panel d. j, Evaluation of chronic analgesic tolerance by hot plate assay. Data are from 7 male mice per genotype. k, Quantification of analgesic efficacy reduction as a difference in the MPE between session 1 and 10 from panel j. Data are from 7 male mice per genotype. l, Evaluation of opioid induced hyperalgesia by hot plate assay. Baseline response latencies of mice receiving repeated fentanyl injections (in panel j) prior to drug administration. m, Quantification of opioid induced hyperalgesia as a difference in the baseline latencies between session 1 and 10 from panel l. Data are from 7 male mice per genotype. n, Evaluation of acute analgesic tolerance by hot plate assay. Mice received repeated morphine injections (20 mg/kg) 120 minutes apart. Data are from 5 WT and 7 Ptchd1 KO mice. o, Quantification of analgesic efficacy reduction as a difference in the MPE between session 1 and 2 from panel n. Data are from 5 WT and 7 Ptchd1 KO mice. p, Evaluation of analgesia by tail immersion assay after intrathecal administration of morphine. Data are from 9 WT and 5 Ptchd1 KO mice. q, Evaluation of analgesic tolerance by tail immersion assay. Mice received repeated intrathecal morphine injections (3.5 μg) over 5 days. Data are from 9 WT and 5 Ptchd1 KO mice. r, Quantification of analgesic efficacy reduction as a difference in the MPE between session 1 and 5 from panel q. Data are from 9 WT and 5 Ptchd1 KO mice. For panel a, d, and q, statistical comparisons were performed by 2-way ANOVA and Šídák post-hoc test. For panel c, e, g, i, j, l, and p, statistical comparisons were performed by 2-way ANOVA and Bonferroni’s post hoc comparison. For panels f, h, and n, statistical comparisons were performed by multiple unpaired two tailed Student’s t-test with Bonferroni’s post hoc. For b, k, m, o, and r, statistical comparisons were performed by unpaired two tailed Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Mean values with S.E.M. errors are shown. MPE, maximum possible effect.

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