Vascular responses with cold (4 °C) water treatment in mice pre-treated with combined TRPA1 antagonist A967079 (100 mg kg–1) and TRPM8 antagonist AMTB (10 mg kg–1), or vehicle control (Veh - 10 % DMSO, 10 % Tween in saline) i.p. 30 min before cold treatment. (a–c) % change in hindpaw blood flow from baseline to 0–2 min following cold treatment (maximum vasoconstriction) in mice treated with combined antagonist (a) A967079+ AMTB, (b) A967079, and (c) AMTB. (d-f) Maximum vasoconstriction caused by cold water treatment in mice treated with combined antagonist (d) A967079+ AMTB, (e) A967079, and (f) AMTB normalised against vehicle treated mice. (n = 5–10) (g–h) RT-PCR CT analysis shows fold change of (g) Trpa1 and (h) Trpm8 normalised to three housekeeping genes in DRG. (n = 14–15) (i) Representative western blot of TRPM8 in DRG of young and aged mice and densitometric analysis normalised to Tubulin (Y = young, A = aged) (n = 9). All results are shown as mean ± s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001. (Two-way ANOVA with Tukey’s post hoc test or Student’s t-test).