Research on the Molecular Mechanism of Crayfish prx 6 in the Process of Defending against Staphylococcus aureus Infection
[Objective]To unravel the molecular intricacies associated with the immune modulatory role of the crayfish prx 6 gene in response to Staphylococcus aureus infection and to expound its function in the orchestration of innate immunity in crayfish,this investigation was carried out.[Method]The study pivoted on the prx 6 gene and harvested total RNA from diverse unstimulated crayfish tissues.The RT-qPCR was utilized to pinpoint the tissue-specific distribution patterns of this gene.Following immune provocation by S.aureus,the prx 6 gene expression dynamics within crayfish's immune-centric tissues were monitored.The research further endeavored to gauge the relative expression of immune executor genes,specifically,Pc-crustin 3,Pc-crustin 4,Pc-ALF 9,and Pc-lectin 1 in the hepatopancreas after RNAi-mediated silencing of prx 6,in the wake of S.aureus infection.Additionally,a quantitative appraisal of the bacterial burden in crayfish hemolymph was undertaken.[Result]The investigations underscored an amplified expression of the prx 6 gene within the hepatopancreas as compared to other tissues.Post S.aureus infection,the prx 6 gene expression showcased substantial augmentation in the hepatopancreas,hemocytes,and intestinal tissues,with a preliminary surge of expression detected in gill tissues.In terms of survival rates,a pronounced decrement in crayfish vitality was observed after prx 6 knockdown,relative to dsGFP+S.aureus controls.To elucidate the key factors linked to escalated mortality,the expression strata of antimicrobial peptide genes integral to bacterial defense were quantified.The outcomes revealed a significant downturn in the expression of Pc-crustin 3,Pc-crustin 4,Pc-ALF 9,and Pc-lectin 1 within the hepatopancreas,seemingly tied to prx 6 repression.Intriguingly,assessment of hemolymph bacterial loads unveiled a substantial elevation in bacterial load in the prx 6 knockdown group as set against controls.[Conclusion]The prx 6 gene in crayfish plays a pivotal role in the antibacterial innate immune response through modulating the expressions of antimicrobial peptide genes and enhancing the clearance capabilities of bacteria from the hemolymph.
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