Fig. 4

Schwann cell TRPA1 modulates painful behaviours in oxaliplatin-induced CIPN. (a) Time-dependent mechanical allodynia induced by oxaliplatin (Oxa, 3 mg·kg?1, i.p.) or vehicle (Veh) in Plp1+-Trpa1 and control mice (n = 8 animals per group). (b) Representative images and cumulative data of 4-HNE immunostaining in mouse sciatic nerve tissue in Plp1+-Trpa1 and control mice after Oxa (3 mg·kg?1, i.p.) or Veh (scale bar: 50 ?m) (n = 5 animals per group, yellow dashes represent the edge of sciatic nerve). (c) Time-dependent mechanical allodynia induced by oxaliplatin (Oxa, 3 mg·kg?1, i.p.) or Veh in Adv+-Trpa1 and control mice (n = 8 animals per group). (d) Schematic representation of zebrafish larvae body section and representative images of the lateral view of 5 days post fertilization (dpf) zebrafish larvae stained with anti-acetylated tubulin (?-tubulin, axons marker) and SOX10 (Schwann cells marker) (scale bar: 20 ?m). (e) Representative images of the lateral view of RNAscope images of TRPA1b mRNA, SOX10 and ?-tubulin in 5 dpf zebrafish larvae. (f) Schematic representation of pTol2-MBP-TurboGFP-dre-trpa1_b[shRNA] plasmid. (g) Schematic representation of zebrafish larvae body section and representative images of SOX10 immunostaining and turbo GFP protein in 5 dpf MBP?-trpa1b zebrafish larvae (scale bar: 10 ?m). (h,i) Writhing behaviour and light touch response in 24?27 post fertilization (hpf) MBP+-trpa1b or control zebrafish larvae exposed to AITC (500 ?M) or Veh (n = 20 larvae per group). (j) H2O2 levels in 5 dpf MBP+-trpa1b or control zebrafish larvae homogenates exposed to AITC (10 ?M) or Veh (n = 5 larvae per group). (k,l) Time-dependent and cumulative data of the locomotor activity of 5 dpf MBP+-trpa1b or control zebrafish larvae exposed to a subthreshold dose of H2O2 (100 ?M) and AITC (0.01 ?M) or Veh following incubation with a subthreshold dose of Oxa (1 ?M) (n = 12 larvae per group). Mean ± SEM. *P < 0.05 (vs. Control-Veh-Oxa in a); #P < 0.05 (vs. Control-Oxa in a) one-way and two-way ANOVA and Bonferroni post hoc test.