Identification, characterization, and functional analysis of Toll and ECSIT in Exopalaemon carinicauda

Toll and evolutionary conserved signaling intermediate in Toll pathways (ECSIT) are two essential molecules in Toll/Toll-like receptor (TLR)-mediated signaling pathway. In this study, Toll and ECSIT (named as EcToll and EcECSIT) were identified for the first time from Exopalaemon carinicauda. EcToll mRNA transcripts were high expressed in hemocytes and gill, and EcECSIT was mainly expressed in gill. The expression levels of EcToll and EcECSIT in gills both responded rapidly to Vibrio parahaemolyticus and WSSV stimulations and three types of antimicrobial peptide (AMP) genes were significantly up-regulated by challenge with V. parahaemolyticus. Knockdown of EcToll or EcECSIT increased the sensitivity of E. carinicauda to V. parahaemolyticus challenge and double knockdown of both EcToll and EcECSIT significantly suppressed the bacterial clearance ability of E. carinicauda in vivo. Furthermore, suppressing EcToll restrained the upregulation of EcECSIT and AMPs and suppressing EcECSIT impaired expression of AMPs by V. parahaemolyticus injection, which indicated that EcToll restricted V. parahaemolyticus infection through activating EcECSIT to induce AMPs. This study provides valuable information about the function of Toll-ECSIT pathway in the innate immunity in crustacean.