Crayfish (Procambarus clarkii) TRPA1 is required for the defense against Aeromonas hydrophila infection under high temperature conditions and contributes to heat sensing

Temperature is an important environmental factor influencing immune responses of crayfish. However, the mechanism underlying how temperature affects immune responses remains unclear. Here, we identified an ortholog of the transient receptor potential ankyrin subtype 1 (TRPA1), a temperature sensor of Drosophila, from Procambarus clarkii (PcTRPA1-1). Its expression was induced by high temperature and challenge with heat-killed A. hydrophila at high temperature, but not at lower temperature. PcTRPA1-1 silencing led to increased mortality of crayfish challenged with live A. hydrophila at high temperature (32 degrees C), but had no statistically significant effect on crayfish mortality at 24 degrees C. This suggests that PcTRPA1-1 is involved in the immune responses of crayfish at high temperature as a potential temperature sensor. Further assay exhibited that PcTRPA1-1 silencing affected immune responses of crayfish, including increase of lipid peroxidation, reduction of total antioxidant capacity, decreased phenoloxidase activity and disruption of circadian rhythm of total hemocyte count entrained by temperature cycles. PcTRPA1-1 silencing also decreased the expression of PcHSP70 and PcHSP90 which are responsive to heat stimuli and bacterial challenge. The results collectively indicate that TRPA1 contributes to heat sensing of crayfish and is required for crayfish defense against bacterial infection.