| The Janus kinase/signal transducers and activators of transcription(JAK/STAT)signalling pathway plays a significant role in immune responses to pathogens.In invertebrates,three core components(Domeless,Hopscotch and STAT92E)of the JAK/STAT pathway were first identified in Drosophila melanogaster and had been confirmed to have immune-related functions.However,it is unclear whether the JAK/STAT signaling pathway affects the innate immune response and characterization in crustaceans.Therefore,we explored whether the JAK/STAT signaling pathway has immune-related functions in Eriocheir sinensis which is an important aquatic economic species in China.The results can help us better understand the role of innate immunity in crustacean and provide a theoretical basis for solving the bacterial diseases of Eriocheir sinensis and other crustacean economic species.In the present study,we clone and descript DOME(EsDOME),JAK(EsJAK)and STAT(EsSTAT)from Eriocheir sinensis.EsDOME is 1299 amino acid protein encoded by 4200 bp open reading frames.EsJAK is 1110 amino acid protein encoded by 3333 bp open reading frames.And EsSTAT is 794 amino acid protein encoded by 2385 bp open reading frames.We found that all three share specific domains with corresponding functions by using bioinformatics analysis.To explore whether the JAK/STAT signaling pathway is potentially linked to immune responses,we used quantitative real-time PCR to detect it.The result showed that EsDOME,EsJAK and EsSTAT were highly expressed in different immune-related tissues including the gill,hepatopancreas and hemocytes.Moreover,after bacterial challenge,Es DOME,EsJAK and EsSTAT were all upregulated significantly in hemocytes.In addition,after interfering with the EsDOME gene and inhibiting the Es STAT protein,we found that the expression of the corresponding antibacterial peptides decreased after being stimulated by bacteria.We conjecture that the JAK/STAT signalling pathway positively regulates antimicrobial peptides expression during the antibacterial immune response.This study completed the construction of the basic framework of JAK/STAT signaling pathway in Eriocheir sinensis and determined its function of regulating antimicrobial peptides expression to affect the antibacterial activity in innate immunity,which provided an important reference for the study of crustacean immune-related signaling pathways. |
