Characterization of microRNAs by deep sequencing in red claw crayfish Cherax quadricarinatus haematopoietic tissue cells after white spot syndrome virus infection.
White spot syndrome virus (WSSV) is one of the most prevalent and widespread viruses in both shrimp and crayfish aquaculture. MicroRNAs (miRNAs) are crucial post-transcriptional regulators and play critical roles in cell differentiation and proliferation, apoptosis, signal transduction and immunity. In this study, miRNA expression profiles were identified via deep sequencing in red claw crayfish Cherax quadricarinatus haematopoietic tissue (Hpt) cell cultures infected with WSSV at both early (i.e., 1 hpi) and late (i.e., 12 hpi) infection stages. The results showed that 2 known miRNAs, namely, miR-7 and miR-184 play key roles in immunity. Meanwhile, 106 novel miRNA candidates were predicted by software in these combined miRNA transcriptomes. Compared with two control groups, 36 miRNAs showed significantly different expression levels after WSSV challenge. Furthermore, 10 differentially expressed miRNAs in WSSV-exposed Hpt cells were randomly selected for expression analysis by quantitative real-time RT-PCR. Consistent with the expression profiles identified by deep sequencing, RT-PCR showed a significant increase or decrease in miRNA expression in Hpt cells after WSSV infection. Prediction of targets of miRNAs such as miR-7, cqu-miR-52, cqu-miR-126 and cqu-miR-141 revealed that their target genes have diverse biological roles, including not only immunity but also transcriptional regulation, energy metabolism, cell communication, cell differentiation, cell death, autophagy, endocytosis and apoptosis. These results provide insight into the molecular mechanism of WSSV infection and highlight the function of miRNAs in the regulation of the immune response against WSSV infection in crustaceans.
State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China; School of Marine Sciences, Ningbo University, Ningbo, Zhejiang 315211, PR China.
Meng-Ru
MR
Zhao
State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.
Chuang
C
Meng
State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.
Xiao-Lu
XL
Xie
School of Marine Sciences, Ningbo University, Ningbo, Zhejiang 315211, PR China. Electronic address: lichenghua@nbu.edu.cn.
Cheng-Hua
CH
Li
State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China; Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources, Xiamen University, State-Province Joint Engineering Laboratory of Marine Bioproducts and Technology, Xiamen 361102, Fujian, PR China. Electronic address: Haipengliu@xmu.edu.cn.
Hai-Peng
HP
Liu
D000818:Animals; D003400:Astacoidea / Q000235:genetics* / Q000276:immunology / Q000821:virology*; D059014:High-Throughput Nucleotide Sequencing; D007113:Immunity, Innate; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D059467:Transcriptome; D046848:White spot syndrome virus 1 / Q000502:physiology*
D035683:MicroRNAs
Crayfish haematopoietic tissue; Innate immunity; MicroRNA; White spot syndrome virus