Fig. 4: Blocking CX3CL1-CX3CR1 signaling in the CeA prevents the extinction of anxiety-like behaviors following acute stress.

Representative images (a) and quantitative analyses (b) for Western blot of CX3CL1 in the CeA (n = 5 mice per group). c Representative images of Iba1 immunostaining and three-dimensional reconstruction of microglia in the CeA. Scale bars, 40 μm (overview) and 20 μm (inset and rendering). d The ARS-treated mice showed inhibited microglial activation after treated with JMS-17-2 (n = 6 mice per group). Representative images (e) and quantification (f) of Iba1⁺ microglia (green) containing GAD65/67⁺ puncta (red) in the CeA (n = 10 cells from five mice per group). Scale bars, 10 μm (overview) and 5 μm (inset and rendering). Representative images of neuronal dendrites (g) and quantification of spine numbers (h) (n = 6 mice per group; h, F_1,20 = 10.16, p = 0.0046). Scale bars, 10 μm. Summarized data of entries and the time spent in the central area and open arms of OFT (i) and EPM (j) (i left, F_1,31 = 11.51, p = 0.0019; right, F_1,31 = 11.54, p = 0.0019; j left, F_1,31 = 11.24, p = 0.0021; right, F_1,31 = 13.51, p = 0.0009). Raster plots and typical traces (k) and the quantitative data (l) of the spontaneous firings of CeA^GABA neurons (n = 27 cells from six mice per group; l, F_1,104 = 3.214, p = 0.0759). m qPCR of Nf-κb in the CeA (n = 5 mice per group). Summarized data of entries and the time spent in the central area and open arms of OFT (n) and EPM (o) (n = 8 mice per group). p qPCR of Cx3cl1 in the CeA (n = 5 mice per group; F_1,16 = 15.63, p = 0.0011). Significance was assessed by two-way repeated-measures ANOVA with post hoc comparison between groups in (b, h, i, j, l, n, o, p), two-tailed unpaired Student’s t-test in (d, f, m). All data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001; n.s., not significant. See also Supplementary Data 1. Source data are provided as a Source Data file.