Grass carp reovirus (GCRV), a member of the Aquareovirus genus in the Reoviridae family, is considered the most pathogenic aquareovirus. However, its productive viral entry pathways remain largely unclear. Using a combination of quantum dot (QD)-based live-virus tracking and biochemical assays, we found that extraction of cellular membrane cholesterol with methyl-β-cyclodextrin (MβCD) and nystatin strongly inhibited the internalization of GCRVs, and supplementation with cholesterol restored viral infection. In addition, the entry of the virus was restrained by genistein, an inhibitor known to block caveolar endocytosis. Subsequent real-time tracking experiments revealed that the QD-labeled GCRV particles were colocalized with caveolin-1, and transfection of cells with dominant-negative mutant (caveolin-1 Y14F) significantly reduced GCRV infection. In contrast, no effects on virus infection were detected when the clathrin-mediated endocytosis or the macropinocytosis inhibitors were used. Our results collectively suggest that aquareoviruses can use caveolae/raft-mediated endocytosis as the primary entry pathway to initiate productive infection.

草鱼呼肠孤病毒（GCRV），呼肠孤病毒科中的水痘病毒属家族，被认为是最具致病性的水痘病毒。但是，其生产性病毒进入途径仍不清楚。使用基于量子点（QD）的活病毒跟踪和生化分析相结合，我们发现用甲基-β-环糊精（MβCD）和制霉菌素提取细胞膜胆固醇会强烈抑制GCRVs的内在化，并补充胆固醇病毒感染。此外，该病毒的进入受到染料木黄酮的抑制，染料木黄酮是一种已知的阻断海绵体内吞作用的抑制剂。随后的实时跟踪实验显示，QD标记的GCRV颗粒与小窝蛋白1共定位，并且用显性阴性突变体（caveolin-1 Y14F）转染细胞可显着减少GCRV感染。相比之下，当使用网格蛋白介导的内吞作用或巨胞饮抑制剂时，未检测到对病毒感染的影响。我们的结果共同表明，水痘病毒可以利用海绵体/筏介导的内吞作用作为引发生产性感染的主要进入途径。