title: Methods to Study Autophagy in Zebrafish.
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.
authors.affiliation: Eötvös Loránd University, Budapest, Hungary.
authors.forename: E
authors.initials: E
authors.lastname: Fodor
authors.affiliation: Eötvös Loránd University, Budapest, Hungary.
authors.forename: T
authors.initials: T
authors.lastname: Sigmond
authors.affiliation: Eötvös Loránd University, Budapest, Hungary.
authors.forename: E
authors.initials: E
authors.lastname: Ari
authors.affiliation: Eötvös Loránd University, Budapest, Hungary.
authors.forename: K
authors.initials: K
authors.lastname: Lengyel
authors.affiliation: Eötvös Loránd University, Budapest, Hungary.
authors.forename: K
authors.initials: K
authors.lastname: Takács-Vellai
authors.affiliation: Eötvös Loránd University, Budapest, Hungary. Electronic address: mvarga@ttk.elte.hu.
authors.forename: M
authors.initials: M
authors.lastname: Varga
authors.affiliation: Eötvös Loránd University, Budapest, Hungary. Electronic address: vellai@falco.elte.hu.
authors.forename: T
authors.initials: T
authors.lastname: Vellai
mesh_terms: D000818:Animals; D001343:Autophagy; D015153:Blotting, Western / Q000379:methods; D064113:CRISPR-Cas Systems; D005786:Gene Expression Regulation; D020868:Gene Silencing; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D016296:Mutagenesis; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D029961:Zebrafish Proteins
keywords: AUTEN (autophagy enhancer); Autophagy; CRISPR-Cas9; LC3B; Morpholinos; Regeneration; Reporters; Zebrafish