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. 2023 Oct 25;14:6696. doi: 10.1038/s41467-023-42283-6

Fig. 5. The level of NADA is decreased in the TRN after CSD.

Fig. 5

a–b Targeted metabolomics analysis of the TRN. Mice in the CSD and sham groups (n = 8 mice). a Volcano plot of metabolites. Differential abundant metabolites were identified using a fold change of 2 and p value (t test) <0.05 as cutoffs. b Relative abundance of NADA (n = 8 mice, Sham vs CSD p = 0.0132). *p < 0.05, two-sided t test. c, d Expression of the CB1 receptor in the TRN. c The picture is representative of six independently stained samples. d TRPV1 expression was negligible in the TRN. Anti-PV, anti-TRPV1, and anti-CB1 receptor antibodies were used. e–g Decreased CB1 receptor activity after CSD. AAV-hSyn-GRAB-eCB2.0 was injected into the TRN (n = 6 mice). Four weeks were allowed for virus expression. Mice were then subjected to CSD. CB1 receptor activity was examined on days 0 and 5. e Diagram of the virus expression and fiber photometry experiments. The image was obtained after staining with an anti-GFP antibody. f Representative traces of CB1 receptor activity. g Area under the curve of TRN CB1 receptor activity traces (n = 6 mice, Day 0 vs day 5 CSD p = 0.0014). **p < 0.01, two-sided paired t test; NS: p > 0.05. Source data are provided as a Source Data file. NADA N-arachidonoyl dopamine, PV parvalbumin, CB1 cannabinoid receptor type 1, TRPV1 transient receptor potential vanilloid 1.