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Generation of transgenic zebrafish with liver-specific expression of EGFP-Lc3: A new in vivo model for investigation of liver autophagy

机译:具有EGFP-Lc3肝脏特异性表达的转基因斑马鱼的生成:研究肝脏自噬的新体内模型

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摘要

Transgenic expression of GFP-Lc3 is a useful tool for an in vivo model to monitor the formation of autophagosomes during the autophagy process. So far, two transgenic animals (mice and zebrafish) with expression of GFP-Lc3 have been reported. Liver is one of the most important organs for autophagy research. Here, we generated a transgenic zebrafish line with liver-specific EGFP-Lc3 expression. By exposing transgenic larvae to the autophagy inducer, Torin1, we observed a substantial increase in the number of EGFP-Lc3 puncta in the liver as well as the increase of Lc3-II protein. Notably, addition of a chloroquine (CQ) led to further increase of EGFP-Lc3 puncta in liver cells due to the blockage of lysosomal function and degradation stage of autophagy. Thus, the newly established transgenic line will be a useful in vivo model to investigate liver autophagy, and, in particular, the involvement of autophagy in basic biology and diseases in the liver.
机译:GFP-Lc3的转基因表达是一种用于体内模型的工具,可在自噬过程中监测自噬体的形成。迄今为止,已经报道了两只表达GFP-Lc3的转基因动物(小鼠和斑马鱼)。肝是自噬研究最重要的器官之一。在这里,我们生成了具有肝特异性EGFP-Lc3表达的转基因斑马鱼品系。通过将转基因幼虫暴露于自噬诱导剂Torin1,我们观察到肝脏中EGFP-Lc3点的数量显着增加以及Lc3-II蛋白的增加。值得注意的是,由于溶酶体功能的阻断和自噬的降解阶段,添加氯喹(CQ)导致肝细胞中EGFP-Lc3点进一步增加。因此,新建立的转基因品系将是用于研究肝脏自噬,尤其是自噬与肝脏基本生物学和疾病有关的体内模型。

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