Fig. 7

Insufficient activation of CaMKIIα neurons in the LPBN on Day 2 after CFA injection. A–E In vivo calcium imaging of CaMKIIα neurons in the LPBN at different time points. A Experimental design. AAV-CaMKIIα-GCaMP6s was injected into the LPBN 21 days before in vivo calcium imaging, and 40-min in vivo calcium imaging was performed 1 day before CFA injection (baseline), the day of CFA injection (CFA 0 d) and Day 2 after CFA injection (CFA 2 d) in awake mice. The recording was continued for 40 min at the same time each day and the recording began 10 min before the start of CFA injection on CFA 0 d. B Quantification of GCaMP6s signals at baseline, CFA 0 d and CFA 2 d in awake mice via AUC of ΔF/F during 0–30 min. C–E Average GCaMP6s signal and heatmap of ΔF/F (%) for all individual mice at baseline, CFA 0 d and CFA 2 d (n = 4 mice). F, G Graphs depicting quantification of c-Fos⁺ neurons in the LPBN and representative images following saline or CFA injection on the day of injection (0 d) and 2 d post injection (n = 9 sections from three mice per group). H Time spent licking the paw following saline or CFA injection on the day of injection (0 d) and 2 d post injection. The recording was continued for 30 min at the same time each day and was immediately performed following CFA injection on CFA 0 d. I Time spent licking paw following CFA injection CFA 0 d and CFA 2 d displayed in 5-min time bins. Data are mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. One-way ANOVA followed by the Tukey’s multiple comparisons test in B. Two-way ANOVA followed by the Sidak’s multiple comparisons test in G-I