Figure 8

8-Br-cAMP-sensitization of TRPA1 Requires PKA-phosphorylation Site. Mustard oil (MO, 50 μM)-activated Ca²⁺ mobilization was monitored from DRG sensory neurons harvested from TRPA1 KO mice and transfected with cDNA corresponding to green fluorescent protein (eGFP-N1, GFP) and either pcDNA3 (empty vector, E.V., A), mouse TRPA1 WT (TRPA1, B), mouse TRPA1 S87A (S87A, C), mouse TRPA1 S179G (S179G, D), mouse TRPA1 S318A (S318A, E), or mouse TRPA1 S1101G (S1101G, F). For 5 min prior to recordings, DRG were pre-incubated for 5 min in vehicle (ddH2O) or 8-Br-cAMP (10 μM), then DRG were challenged with MO (30 sec) and Ca²⁺ accumulation was measured. Cumulative traces are displayed in panels A–F, quantitative measurements from multiple neurons displayed in panel G. *p < 0.05; **p < 0.01; ns = no significance; n = 15–39 MO responsive cells per TRPA1 cDNA transfection; trace significance determined by two-way ANOVA with Bonferroni posthoc analysis and quantified data significance determined by unpaired student’s t test.