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| Inhibition of TiLV Replication by AT2-R in Nile Tilapia Oreochromis niloticus |
| WEN Jing1,2, ZHENG Shucheng2, KE Zishan2, WANG Yingying2, LI Yingying2, MO Xubing2, ZHANG Defeng2, YIN Jiyuan2, ZHOU Wenli1, WANG Qing2 |
1. College of Fisheries, Tianjin Agricultural University, Tianjin 300384, China;
2. Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture, Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China |
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Abstract In order to explore the function of type-2 angiotensin Ⅱ receptor (AT2-R) of Nile tilapia Oreochromis niloticus in the process of tilapia lake virus (TiLV) infection, the complete open reading frame (ORF) sequence of AT2-R gene of Nile tilapia AT2-R (OnAT2-R) was cloned by reverse transcription PCR (RT-PCR) and the tissue distribution and phase expression were analyzed in OnAT2-R by real-time fluorescence quantitative PCR (qRT-PCR). Meanwhile, the recombinant eukaryotic expression vector pEYFP-OnAT2-R was constructed for subcellular localization analysis in tilapia brain (TiB) cells which were infected with TiLV after overexpression of OnAT2-R gene, and the relative expression of TiLV-S8 segment was detected by qRT-PCR. The results showed that the OnAT2-R gene ORF sequence of the Nile tilapia was found to be 1338 bp in length, encoding 445 amino acids with a 7-transmembrane region. The OnAT2-R gene was found in all tissues examined in the Nile tilapia, with the maximal expression level in liver, followed by muscle and skin, and the minimal expression level in spleen and stomach. The relative expression level of OnAT2-R gene in the liver was shown to be increased first and then decreased in the Nile tilapia with TiLV infection, while the relative expression of OnAT2-R gene in the muscle was increased and then decreased, followed by increased again. Furthermore, the pEYFP-OnAT2-R was successfully constructed and the subcellular localization analysis showed that OnAT2-R protein was mainly localized on the cell membrane. The mRNA level of TiLV-S8 segment was significantly reduced in TiB cells overexpressed with OnAT2-R gene followed by TiLV infection. The finding indicates that OnAT2-R is a potential cell surface molecular receptor located on the cell membrane and has the effect on inhibition of TiLV replication, which will provide theoretical references with further in-depth research on pathogenesis of TiLV infection and antiviral mechanism of tilapia.
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Received: 23 February 2023
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