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. 2022 May 17;219(6):e20212019. doi: 10.1084/jem.20212019

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© 2022 Hasebe et al.

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Figure 1. — Activated sensory pathways are involved in bilateral ankle joint inflammation. (A and B) Clinical scores of the left (A) and right (B) ankles of F759 mice deafferented or sham-operated at L5 DRG of the left side. IL-17A and IL-6 (0.1 μg each) were injected into both ankles on days 0, 1, and 2 (n = 4 per group). (C) CD11b+MHC II+ cells in the ankle joints of F759 mice on day 3 after IL-17A and IL-6 or saline injections. IL-17A and IL-6 or saline (1 μg each) were injected into the left ankle joint on days 0, 1, and 2. Green, MHC II. Magenta, CD11b. Blue, nuclei. Bars, 1 mm. Arrowheads show CD11b+MHC II+ cells. Experiments were performed at least three times independently; representative data are shown. (D) Clinical scores of the right ankle joint of F759 mice after IL-17A and IL-6 injections at a low dose (0.01 μg each) in the right ankle joint and high dose (1 μg each) or saline injection in the left ankle joint on days 0, 1, and 2 (n = 14 per group). Mean scores ± SEM are shown. P values were calculated using the Wilcoxon rank-sum test (*, P < 0.05; **, P < 0.01). (E and F) IL-17A and IL-6 (1 μg each) or saline was injected into the left ankle joint on days 0, 1, and 2. c-fos expression in ipsilateral (E) or contralateral (F) L3, L4, L5, and L6 DRGs on day 3 using real-time PCR (n = 5–6 per group). (G and H) IL-17A and IL-6 (1 μg each) or saline was injected into the left ankle joint on days 0, 1, and 2, followed by staining of c-fos, Nav1.8, and TRPV1 in the ipsilateral (G) or contralateral (H) L5 DRG on day 3. Magenta, c-fos. Green, Nav1.8 or TRPV1. Blue, nuclei. Yellow arrowheads show c-fos+Nav1.8+TRPV1− neurons. White arrows show c-fos+Nav1.8+TRPV1+ neurons. The number of c-fos+, Nav1.8+TRPV1−, Nav1.8+TRPV1+, and Nav1.8−TRPV1+ cells was counted (n = 3 per group). Bar graphs show the percentage of c-fos+ cells in the total number of Nav1.8+TRPV1−, Nav1.8+TRPV1+, or Nav1.8−TRPV1+ neurons. Bar, 100 µm. Experiments were performed three times independently. The data are shown as means ± SEM. P values were calculated using Student’s t-tests (*, P < 0.05; **, P < 0.01). Diagrams depicted above or to the left of the graphs illustrate the experimental settings. L, left ankle; R, right ankle. Arrows indicate cytokine or saline injections, and blue circles indicate the ankle joint assessed for arthritis (A–D) or the DRG examined (E–H).