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1.
Dev Comp Immunol ; 107: 103642, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32061940

RESUMO

The respiratory glycoprotein, hemocyanin (HMC) has multiple immune-related functions, including antiviral activity. In this study, in silico methods were used to predict seven miRNAs targeting Litopenaeus vannamei HMC (LvHMC), out of which miR-589-5p was selected for further investigation because of its role in immune response. Transcript levels of miR-589-5p were ubiquitously distributed in all shrimp tissues examined, and significantly induced in hemocytes and hepatopancreas upon challenge with white-spot syndrome virus (WSSV) as well as by marine bacterial pathogens, which suggest that miR-589-5p is involved in shrimp immune response to pathogens. Morever, using Drosophila S2 cells stably overexpressing EGFP-LvHMC, flow cytometry and dual luciferase reporter assays, miR-589-5p was shown to significantly inhibit the in vitro expression of LvHMC. In addition, in vivo knockdown of miR-589-5p using antagomir-589-5p resulted in significant down-regulation in LvHMC expression, while overexpression of miR-589-5p using agomir-589-5p decreased the level of LvHMC expression in shrimp hemocytes and hepatopancreas. Further, the increased expression of miR-589-5p resulted in high shrimp mortality following WSSV challenge, coupled with an increase in the number of WSSV copies in hemocytes and hepatopancreas. These results suggest that miR-589-5p is involved in shrimp immune response to WSSV by negatively regulating the expression of LvHMC.


Assuntos
Proteínas de Artrópodes/metabolismo , Infecções por Vírus de DNA/imunologia , Hemocianinas/metabolismo , Hemócitos/fisiologia , MicroRNAs/genética , Penaeidae/imunologia , Vírus da Síndrome da Mancha Branca 1/fisiologia , Animais , Proteínas de Artrópodes/genética , Linhagem Celular , Drosophila , Regulação da Expressão Gênica , Hemocianinas/genética , Hepatopâncreas , Imunidade Inata/genética
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