Fig. 5: NOPLight in vivo reports bidirectional endogenous N/OFQ dynamics during consummatory and aversive behaviors.
From: Development of a genetically encoded sensor for probing endogenous nociceptin opioid peptide release
a Fiber photometry schematic. Cartoon of viral injection of NOPLight and fiber implant in VTA of wild-type mice (n = 7 mice). b Cartoon depicting head-fixed cued-sucrose setup and trial structure. c Averaged traces of NOPLight fluorescence following pretreatment with vehicle (veh, green) or 20 mg/kg NOPR antagonist J-113397 (J11, blue), aligned to tone onset (magenta, shaded). Data represented as mean ± SEM. d Average licks made during vehicle or J-113397 sessions (two-tailed Wilcoxon test, p = 0.1649, ns not significant, n = 7 mice). Data represented as mean ± SEM. e Heat maps of NOPLight fluorescence, rows correspond to trials averaged in (c) for vehicle (left) and J11 (right) sessions. f Area under the curve (AUC) for averaged traces from (c), calculated over 5-s intervals surrounding cued-sucrose events. Decrease in NOPLight fluorescence during 10% sucrose access (two-tailed Wilcoxon test, **p = 0.0034, n = 7 mice) is blocked by J11 pretreatment (two-tailed Mann–Whitney test, ##p = 0.0022, n = 7 mice). Data represented as mean ± SEM. g Top: Fiber photometry schematic. Middle: Cartoon depicting fiber implant and viral injection of DIO-GCaMP6m, FLEX-NOPLight, or NOPLight-ctr into the VTA of PNOC-Cre, OPRL1-Cre, or WT mice, respectively. Bottom: Session trial structure. h Left: Averaged trace of pnVTAPNOC GCaMP6m activity during tail lift. Right: AUC for photometry trace calculated over 5-s intervals surrounding tail lift (two-tailed Wilcoxon test, ***p = 0.0002 (5–10 s) or 0.0003 (10–15 s); ****p < 0.0001, n = 4 mice). Data represented as mean ± SEM. i Left: Averaged traces of FLEX-NOPLight (green) and NOPLight-ctr (gray) fluorescence during the tail lift. Right: AUC for photometry traces calculated over 5-s intervals surrounding tail lift. FLEX-NOPLight fluorescence increases during tail lift (two-tailed Wilcoxon test, **p = 0.0034 (0–5 s) or 0.0093 (5–10 s); n = 3 mice), NOPLight-ctr fluorescence remains unchanged (two-tailed Mann–Whitney test, #p = 0.0264 (0–5 s) or 0.0326 (5–10 s); n = 8 mice, NOPLight-ctr; 3 mice, FLEX-NOPLight). Data represented as mean ± SEM. VITI variable inter-trial interval.
