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. 2024 Mar 6;121(11):e2312136121. doi: 10.1073/pnas.2312136121

Fig. 3.

Fig. 3.

The pharynx→NJP→NTSNE circuit controls pharyngeal inflammation–induced anxiety. (A) Schematic of the viral injection and wireless optogenetics. (B) Typical image showing the infusion site within the NTS. (Scale bar, 100 μm.) (C) Typical images showing the infusion site within NJP superganglia. (Scale bars, 100 μm.) The white boxes depict the areas shown in the boxes of the NJP superganglia. (Scale bars, 10 μm.) L: Left; R: Right. (D) Representative heatmaps of the travel trajectory and summarized data for the EPM test with EYFP and eNpHR-EYFP CFA mice. (n = 10 mice per group; Left, t18 = 3.578, P = 0.0022; Right, t18 = 6.934, P < 0.0001). (E) Representative heatmaps of the travel trajectory and summarized data for the OF test with EYFP and eNpHR-EYFP CFA mice. (n = 10 mice per group; Left, t18 = 8.093, P < 0.0001; Middle, t18 = 6.000, P < 0.0001; Right, t18 = 0.8318, P = 0.4164). (F) Representative heatmaps of the travel trajectory and summarized data for the LDB test with EYFP and eNpHR-EYFP CFA mice. (n = 10 mice per group; Left, t18= 5.794, P < 0.0001; Right, t18 = 4.090, P = 0.0007). (G) Schematic of the viral injection and microendoscope imaging in freely moving mice. (H) Representative image showing the expression of GCaMP6m and the GRIN lens tract in the NTS. (Scale bar, 100 µm.) (I) Sample traces (Left) and summarized data (Right) showing the spontaneous Ca2+ transient frequency of NTSNE neurons from saline and CFA mice. (Saline, n = 49 cells; CFA, n = 51 cells; U = 831, P = 0.0036). (J) Schematic of viral injection and chemogenetic inhibition design. (K) Representative image showing the expression of hM4Di-mCherry in the NTS (Left) and predominantly colocalized with DBH antibody (Right). (Scale bars, 100 µm [Left] or 20 µm [Right].) (L) Representative heatmaps of the travel trajectory and summarized data for the EPM test with mCherry and hM4Di-mCherry CFA mice (after injection of CNO; n = 10 mice per group; Left, t18 = 4.591, P = 0.0002; Right, t18 = 6.252, P < 0.0001). (M) Representative heatmaps of the travel trajectory and summarized data for the OF test with mCherry and hM4Di-mCherry CFA mice (after injection of CNO; n = 10 mice per group; Left, t18 = 8.436, P < 0.0001; Middle, t18 = 6.757, P < 0.0001; Right, t18 = 0.03862, P = 0.9696). (N) Representative heatmaps of the travel trajectory and summarized data for the LDB test with mCherry and hM4Di-mCherry CFA mice (after injection of CNO; n = 10 mice per group; Left, t18 = 5.315, P < 0.0001; Right, t18 = 4.551, P = 0.0002). Significance was assessed by Mann–Whitney U test in (I) and two-tailed unpaired Student’s t tests in (D, E, F, L, M, and N). All data are presented as the mean ± SEM. **P < 0.01; ***P < 0.001, n.s., not significant.