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Toll-like receptor 3 regulates Mx expression in rare minnow Gobiocypris rarus after viral infection

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Abstract

Toll-like receptor 3 (TLR3) plays a key role in activating immune responses during viral infection. To study the genes involved in the regulatory function of TLR3 in the rare minnow Gobiocypris rarus after viral infection, a full-length cDNA of TLR3 (GrTLR3) with a splice variant (GrTLR3s) was identified by homologous cloning and RACE techniques. The antiviral effector molecule Mx gene was cloned and partially sequenced. The mRNA expression levels of GrTLR3, GrTLR3s, and Mx were studied in different tissues before and after virus infection by real-time quantitative RT-PCR. The transcripts of all three genes in liver were significantly increased following GCRV infection (P < 0.05). The mRNA levels in liver were upregulated at 24 h post-injection for GrTLR3 and GrTLR3s, and at 12 h for Mx. The upregulated expression levels were several folds for GrTLR3s, tens of folds for GrTLR3, and hundreds of folds for Mx. By semi-quantitative RT-PCR, GrTLR3 and Mx expressed at all the developmental stages, whereas GrTLR3s could only be detected at later developmental stages. Using RNAi and transgenic techniques, GrTLR3 mediated Mx expression but GrTLR3s did not. The time-dependent upregulation of receptor and effector, and the Mx over-expression dependent on TLR3, indicated that GrTLR3 regulated Mx expression in viral infection through a configuration change in rare minnow, and its splice variant did not contribute to the process.

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Acknowledgement

The authors would like to thank Ms. Ming Li and Mr. Renxiang Yan for fish administration and microinjection, and Ms. Shangping Chen and other laboratory staff for technical advice and helpful discussions. We are grateful to Dr. Yonghong Wang from CSIRO for critically reading the manuscript. This study was supported by the National Natural Science Foundation of China (30428024, 30540084 and 30740009), 973 National Basic Research Program of China (2006CB102100 and 2004CB117406), Chinese Academy of Sciences project (KSCX2-YW-N-021), research foundation (08080262, 08080245 and 01140508) from Northwest A&F University in China, China Postdoctoral Science Foundation (20070410298), and the grant from the State Key Laboratory of Freshwater Ecology and Biotechnology (2007FB09).

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Authors and Affiliations

  1. Institute of Hydrobiology, The Chinese Academy of Sciences, Wuhan, 430072, People’s Republic of China

    Jianguo Su, Zuoyan Zhu, Yaping Wang & Wei Hu

  2. Northwest A&F University, Yangling, 712100, People’s Republic of China

    Jianguo Su

  3. School of Biological Sciences, University of Aberdeen, Aberdeen, AB24 2TZ, UK

    Jun Zou

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  1. Jianguo Su
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  2. Zuoyan Zhu
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Correspondence to Zuoyan Zhu.

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Su, J., Zhu, Z., Wang, Y. et al. Toll-like receptor 3 regulates Mx expression in rare minnow Gobiocypris rarus after viral infection. Immunogenetics 60, 195–205 (2008). https://doi.org/10.1007/s00251-007-0264-0

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