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Immunofluorescent Labeling of Plant Virus and Insect Vector Proteins in Hemipteran Guts
Published on: May 14, 2021
Endocytic pathway is indicated for white spot syndrome virus (WSSV) entry in shrimp
Zih-Jhan Huang1, Shih-Ting Kang, Jiann-Horng Leu
1Institute of Marine Biology, National Taiwan Ocean University, No. 2, Pei-Ning Road, Keelung 20224, Taiwan, ROC.
Abstract:
The white spot syndrome virus (WSSV) has had a serious economic impact on the global shrimp aquaculture industry in the past two decades. Although research has clarified a lot about its genome and structure, the mechanism of how WSSV enters a cell is still unclear. In this study to determine this mechanism, primary cultured hemocytes were used as an experimental model to observe the process of WSSV entry because the stable shrimp cell lines for WSSV infection are lacking. After labeling virions and endosomes with fluorescent dyes followed by observation with a confocal microscope, the results show that the WSSV colocalizes with early endosomes. Hemocytes are further treated with different endocytic inhibitors, methyl-β-cyclodextrin (MβCD) and chlorpromazine (CPZ). WSSV still can be detected in the hemocytes treated with CPZ, but not in the hemocytes treated with MβCD. Thus, we conclude that WSSV adopts the caveolae-mediated endocytosis to enter the shrimp cell.
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