Fig. 3: LI-cTBUS restores spinal BDNF/KCC2 signaling and inhibitory function to counteract neuropathic pain. | Nature Communications

Fig. 3: LI-cTBUS restores spinal BDNF/KCC2 signaling and inhibitory function to counteract neuropathic pain.

From: Harnessing theta-gamma coupled brainwaves using ultrasound for spinal astrocyte revitalization and sustained neuropathic pain relief in mice

Fig. 3

a Left: Experimental schematic for measuring total BDNF protein in the lumbar spinal cord (whole segment); center: representative immunoblots; right: group quantification. N = 6 independent experiments; data are presented as box-and-whisker plots; one-way ANOVA with Tukey’s post-hoc test, *p = 0.02. b Experimental schematic for measuring KCC2 protein levels in the ipsilateral dorsal lumbar spinal cord; center: representative immunoblots; right: quantification. N = 6 independent experiments; data are presented as box-and-whisker plots; *p = 0.03, ***p = 0.000094, one-way ANOVA with Tukey’s multiple comparison test. c Intrathecal injection of recombinant human BDNF (1 ng/10 μL) transiently lowered the withdrawal threshold, lasting for <3 h, N = 4 (BDNF), N = 5 (Saline); mean ± SEM; **p = 0.003, two-way ANOVA with Tukey’s multiple comparison test. d Timeline for LI-cTBUS (200 Hz, 0.77 W/cm²) applied daily after BDNF injection. Center: time-course of withdrawal threshold; right: PWT change on days 3 and 4. N = 6; data are presented as mean ± SEM and box-and-whisker plots; *p = 0.05, **p = 0.005, two-way ANOVA with Tukey’s multiple comparison test. e Total BDNF protein 5 days after BDNF injection with or without LI-cTBUS. Top: blot; bottom: quantification. N = 4 independent experiments; data are presented as box-and-whisker plots; *p = 0.01; ns: non-significant test. f Perforated patch-clamp recordings from lamina II neurons. Left: GABA-evoked current–voltage (I–V) representative traces; center: representative traces; right: summary of chloride reversal potential (Eanion). PCI caused a right-shifted Eanion that was normalized by LI-cTBUS. Data are presented as mean ± SEM. **p = 0.002, ***p = 0.000002; n = 3–10 (control = 3, PCI = 5, PCI + LI-cTBUS = 10), one-way ANOVA with Tukey’s multiple comparison test. N number of mice, n number of recorded cells. Schematic illustrations were created in BioRender. Sung, Y. (2025) https://BioRender.com/figix4a.

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