title: Methods for assessing autophagy and autophagic cell death.
abstract: Autophagic (or type 2) cell death is characterized by the massive accumulation of autophagic vacuoles (autophagosomes) in the cytoplasm of cells that lack signs of apoptosis (type 1 cell death). Here we detail and critically assess a series of methods to promote and inhibit autophagy via pharmacological and genetic manipulations. We also review the techniques currently available to detect autophagy, including transmission electron microscopy, half-life assessments of long-lived proteins, detection of LC3 maturation/aggregation, fluorescence microscopy, and colocalization of mitochondrion- or endoplasmic reticulum-specific markers with lysosomal proteins. Massive autophagic vacuolization may cause cellular stress and represent a frustrated attempt of adaptation. In this case, cell death occurs with (or in spite of) autophagy. When cell death occurs through autophagy, on the contrary, the inhibition of the autophagic process should prevent cellular demise. Accordingly, we describe a strategy for discriminating cell death with autophagy from cell death through autophagy.
authors.affiliation: INSERM, Unit Apoptosis, Cancer and Immunity, Villejuif, France.
authors.forename: Ezgi
authors.initials: E
authors.lastname: Tasdemir
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Galluzzi
authors.forename: M Chiara
authors.initials: MC
authors.lastname: Maiuri
authors.forename: Alfredo
authors.initials: A
authors.lastname: Criollo
authors.forename: Ilio
authors.initials: I
authors.lastname: Vitale
authors.forename: Emilie
authors.initials: E
authors.lastname: Hangen
authors.forename: Nazanine
authors.initials: N
authors.lastname: Modjtahedi
authors.forename: Guido
authors.initials: G
authors.lastname: Kroemer
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001343:Autophagy / Q000502:physiology*; D016923:Cell Death / Q000502:physiology; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D008247:Lysosomes / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000648:ultrastructure; D010588:Phagosomes / Q000648:ultrastructure