PUBLICATION

Methods to Study Autophagy in Zebrafish

Authors
Fodor, E., Sigmond, T., Ari, E., Lengyel, K., Takács-Vellai, K., Varga, M., Vellai, T.
ID
ZDB-PUB-170227-2
Date
2017
Source
Methods in enzymology   588: 467-496 (Chapter)
Registered Authors
Varga, Máté
Keywords
AUTEN (autophagy enhancer), Autophagy, CRISPR-Cas9, LC3B, Morpholinos, Regeneration, Reporters, Zebrafish
MeSH Terms
  • Animals
  • Autophagy*
  • Blotting, Western/methods
  • CRISPR-Cas Systems
  • Gene Expression Regulation
  • Gene Silencing
  • Microscopy, Electron, Transmission/methods
  • Microscopy, Fluorescence/methods
  • Mutagenesis
  • Polymerase Chain Reaction/methods
  • RNA, Messenger/genetics
  • Zebrafish/genetics
  • Zebrafish/metabolism*
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism
PubMed
28237116 Full text @ Methods Enzymol.
Abstract
Autophagy (cellular self-eating) is a highly regulated degradation process of the eukaryotic cell during which parts of the cytoplasm are delivered into, and broken down within, the lysosomal compartment. The process serves as a main route for the elimination of superfluous and damaged cellular constituents, thereby mediating macromolecular and organellar turnover. In addition to maintaining cellular homeostasis, autophagy is involved in various other cellular and developmental processes by degrading specific regulatory proteins, and contributing to the clearance of intracellular pathogens. The physiological roles and pathological involvement of autophagy can be effectively studied in divergent eukaryotic model systems ranging from yeast to mice. Such a tractable animal modelapplied only recently for autophagy researchis the zebrafish Danio rerio, which also facilitates the analysis of more specific biological processes such as tissue regeneration. In this chapter, we overview the main methods and tools that are used to monitor autophagic structures and to assay autophagic responses in this vertebrate organism. We place emphasis on genetic (functional) approaches applied for exploring novel cellular and developmental roles of the autophagic process.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping