Differential microRNA expression profile of Portunus trituberculatus under florfenicol stress

Abstract

Florfenicol (FLR), a broad-spectrum antibacterial agent, is commonly used in the breeding and cultivation of aquatic animals. microRNAs (miRNAs) control gene expression by mediating post-transcriptional mRNA repression. However, not much has been reported regarding their functions when crustaceans encounter FLR stress. Herein we observed histological alterations and performed small RNA sequencing to characterize differentially expressed miRNAs in the hepatopancreas of Portunus trituberculatus exposed to FLR (80 mg/kg) stress for 3 h and 12 h. Our results indicated irregularities in the tubular structure of hepatopancreas, separation between epithelium and myoepithelial layer, ruptured epithelial cells, and tubule degeneration. Thirty-five (19 up- and 16 downregulated) and 31 (17 up- and 14 downregulated) differentially expressed miRNAs were detected between Group-C and Group-FLR-3 h and between Group-C and Group-FLR-12 h, respectively. Dual luciferase reporter assay was performed for predicting target genes of miR-34 and miR-263b. We found that mir-263b directly inhibited cytochrome C expression at the cell level in vitro. Functional analyses of predicted target genes of differentially expressed miRNAs revealed that FLR stress induced xenobiotic metabolism, immune toxicity, and cell apoptosis, as well as influenced diverse metabolic processes. To our knowledge, this is the first report on the response of P. trituberculatus to FLR stress. Our findings provide evidence that miRNAs participate in cell apoptosis and immune regulation in crabs.

Keywords

• Portunus trituberculatus
• Florfenicol
• MiRNA-seq
• Dual luciferase reporter assay
• Immunity
• Apoptosis