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

Biochem Biophys Res Commun. 2012 Jun 1;422(2):268-73. doi: 10.1016/j.bbrc.2012.04.145. Epub 2012 May 3.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Animals, Genetically Modified*
  • Autophagy / drug effects
  • Autophagy / genetics*
  • Chloroquine / pharmacology
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics
  • Liver / cytology*
  • Liver / metabolism
  • Liver / physiology
  • Mice
  • Microtubule-Associated Proteins / biosynthesis
  • Microtubule-Associated Proteins / genetics
  • Models, Animal
  • Molecular Sequence Data
  • Naphthyridines / pharmacology
  • Up-Regulation
  • Zebrafish / genetics*

Substances

  • 1-(4-(4-propionylpiperazin-1-yl)-3-(trifluoromethyl)phenyl)-9-(quinolin-3-yl)benzo(h)(1,6)naphthyridin-2(1H)-one
  • Map1lc3b protein, mouse
  • Microtubule-Associated Proteins
  • Naphthyridines
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Chloroquine