Fig. 3

Oxaliplatin increases oxidative stress, which targets TRPA1 to induce nociceptive behaviour and calcium response. (a) 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 (α-tubulin) and cumulative data of axon branch density following exposure to oxaliplatin (Oxa, 1–10 μM) or vehicle (Veh) (n = 6 larvae per group). (b) Typical traces and cumulative data of the locomotor activity of 5 dpf zebrafish larvae exposed to Oxa (1–10 μM). (c) Schematic representation of zebrafish larvae body section and representative images of the lateral view of 5 dpf zebrafish larvae injected with Hyper7 mRNA at one-cell stage and cumulative data of HyPer ratio (YFP500/YFP400) fluorescence following exposure to Oxa (10 μM) or Veh and in the presence of reduced glutathione (GSH, 10 μM) (n = 6 larvae per group). (d) Typical traces and cumulative data of the locomotor activity of 5 dpf zebrafish larvae exposed to Oxa (10 μM) or Veh and in the presence of GSH (10 μM) (n = 13–16 larvae per group). (e) Schematic representation of zebrafish larvae body section and representative images of the lateral view of 5 dpf zebrafish larvae injected with Hyper7 mRNA at one-cell stage and cumulative data of HyPer ratio (YFP500/YFP400) fluorescence following exposure to Oxa (10 μM) or Veh and in the presence of A-967079 (A96, 1 μM) (n = 6 larvae per group). (f) Typical traces and cumulative data of locomotor activity of 5 dpf zebrafish larvae exposed to Oxa (10 μM) or Veh and in the presence of A96 (10 μM) (n = 16 larvae per group). (g–i) Typical traces and cumulative data of the locomotor activity of 5 dpf exposed to a subthreshold dose of H2O2 (100 μM) and AITC (0.01 μM) or Veh following incubation with a subthreshold dose of Oxa (1 μM) or Veh and in the presence of A96 (10 μM) or GSH (10 μM) (n = 16 larvae per group). (j) Cumulative data of the calcium response evoked by subthreshold concentration of H2O2 (30 μM) in zTRPA1b-HEK293T cells pre-incubated with Oxa (300 μM) or Veh in the absence or presence of A96 (30 μM) or GSH (30 μM) (cell numbers: Veh-Veh n = 83, Veh-H2O2 n = 78, Veh-Oxa n = 111, Oxa-H2O2 n = 129, Oxa-H2O2-A96 n = 156, Oxa-H2O2-GSH n = 148; n = 3 independent experiments). Mean ± SEM. *P < 0.05, #P < 0.05, one-way ANOVA and Bonferroni post hoc test.