Figure 9

mGluR5-sensitization of TRPA1 Requires PKA and PKC-phosphorylation Sites. 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 mouse TRPA1 WT (TRPA1, A), mouse TRPA1 S87A (S87A, B), mouse TRPA1 S119G (S119G, C), mouse TRPA1 T281A (T281A, D), or mouse TRPA1 T529A (T529A, E). For 5 min prior to recordings, DRG were pre-incubated for 5 min in vehicle (ddH2O) or DHPG (10 μM), then DRG were challenged with MO (30 sec) and Ca²⁺ accumulation was measured. Cumulative traces are displayed in panels A–E, quantitative measurements from multiple neurons displayed in panel F. **p < 0.01; ns = no significance; n = 21–35 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.