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. 2023 Jan 12;15:1006125. doi: 10.3389/fnmol.2022.1006125

Figure 1.

Figure 1

CFA 1D mice display pain and anxiety-like behaviors. (A) Schematic of the experimental procedure for mice injected with CFA and behavioral tests. (B) Left hind paw of mice on day 1 after injection of saline or CFA. (C) CFA-induced changes in pain threshold using the von Frey test (Saline 1D, n = 8 mice; CFA 1D, n = 7 mice; F(1, 13) = 199.7, p < 0.0001). (D,F) Heatmaps showing locomotion traces of CFA 1D mice and Saline 1D mice in the open field test (OFT) and elevated plus maze test (EPMT). (E,G) Summarized data of the time spent in and entries into the central area of the OFT (E, Saline 1D, n = 12 mice; CFA 1D, n = 7 mice; entries into the center in the OFT, t17 = 2.282, p = 0.0357; time in the center in the OFT, t17 = 2.122, p = 0.0488; total distance, t17 = 0.9713, p = 0.3450) and open arms of the EPMT (G, Saline 1D, n = 13 mice; CFA 1D, n = 11 mice; entries into the open arms, t22 = 2.471, p = 0.0217; time in the open arms, t22 = 3.375, p = 0.0027) in the saline 1D or CFA 1D mice. (H) Diagram of crossing Gad2-Cre mice and Ai9 reporter mice. (I) Representative images of GABAergic neurons expressing tdTomato in the CeA from GAD2-tdTOM mice. Scale bar, 200 μm. (J) GABA immunofluorescence were colocalized with tdTomato-labeled neurons within the CeA. Scale bar, 20 μm. (K,L) Sample traces (K) and statistical data (L) for action potential firing recorded from GABACeA neurons (red cells in J) in CFA 1D or Saline 1D mice (n = 24 cells from five mice for Saline1D group; n = 17 cells from four mice for CFA 1D group; F(1, 39) = 18.59, p = 0.0001). (M) Summary of the rheobases of action potentials from GABACeA neurons in Saline 1D or CFA 1D mice (n = 24 cells from five mice for Saline1D group; n = 17 cells from four mice for CFA 1D group; t39 = 3.640, p = 0.0008). (N) The resting membrane potential of the CFA 1D group increased compared with that of the Saline group (n = 24 cells from five mice for the saline group; n = 17 cells from four mice for the CFA 1D group; t39 = 1.828, p = 0.0751). All data are shown as the mean ± SEM, n.s., not significant; * p < 0.05; ** p < 0.01 and *** p < 0.001. Two-way repeated-measures ANOVA with Bonferroni post-hoc analysis for (C,L); unpaired t-test for (E,G,M,N).