Abstract
MicroRNAs (miRNAs) are important signaling regulators that are involved in regulating the innate immunity of crustacean. However, few studies focus on the role of crustacean miRNAs in the cellular immunity have been reported. In this study, we showed that the expression of miR-133 was significantly up-regulated in the mud crab Scylla paramamosain after infection by white spot syndrome virus (WSSV) or Vibrio parahaemolyticus. The anti-miRNA oligonucleotide AMO-miR-133 was used to knock down miR-133 expression in S. paramamosain. The number of WSSV copies increased significantly in WSSV-infected crabs after miR-133 knockdown. Knockdown of miR-133 also enhanced the mortality rates of WSSV-infected and V. parahaemolyticus-infected mud crabs, and it significantly enhanced the expression of the astakine, which was confirmed by real-time quantitative PCR and western blot analysis. The data also indicate that miR-133 may affect hemocyte proliferation in S. paramamosain by regulating astakine expression. miR-133 Knockdown enhanced the apoptosis or phagocytosis of crab hemocytes, and increased the mortality of mud crabs after WSSV or V. parahaemolyticus infection. These results indicate that miR-133 is involved in the host immune response to WSSV and V. parahaemolyticus infection in mud crabs. Taken together, our research provides new insights for the control of viral or vibrio diseases in S. paramamosain.
Highlights
MicroRNAs are highly conserved endogenous small non-coding RNAs, which regulate gene expression at post-transcriptional levels and protein synthesis in eukaryotes [1]. miRNAs play major roles in regulating biological processes, including cell proliferation, cell differentiation, apoptosis, DNA replication, and signal transduction [2,3,4]
Zhu et al identified more than 50 kinds of miRNAs that responded to Vibrio alginolyticus infection in Marsupenaeus microRNA-133 Regulates Innate Immunity japonicus, and the expression levels of some of them were upregulated [7]
To determine whether miR-133 is involved in the response of S. paramamosain to white spot syndrome virus (WSSV) or V. parahaemolyticus infection, hemocytes of healthy, WSSV-infected, and V. parahaemolyticusinfected mud crabs were collected to measure the expression of miR-133
Summary
MicroRNAs (miRNAs) are highly conserved endogenous small non-coding RNAs, which regulate gene expression at post-transcriptional levels and protein synthesis in eukaryotes [1]. miRNAs play major roles in regulating biological processes, including cell proliferation, cell differentiation, apoptosis, DNA replication, and signal transduction [2,3,4]. The expression level of some miRNAs changed in V. parahaemolyticus-infected shrimp (Litopenaeus vannamei), including 47 miRNAs that were up-regulated and 36 miRNAs that were down-regulated [6]. Zhu et al identified more than 50 kinds of miRNAs that responded to Vibrio alginolyticus infection in Marsupenaeus microRNA-133 Regulates Innate Immunity japonicus, and the expression levels of some of them were upregulated [7]. These reports indicate that crustacean miRNAs are associated with the process of Vibrio infection and may play an important role in regulation of innate immunity
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