Methods for assessing autophagy and autophagic cell death

Ezgi Tasdemir, Lorenzo Galluzzi, M. Chiara Maiuri, Alfredo Criollo, Ilio Vitale, Emilie Hangen, Nazanine Modjtahedi, Guido Kroemer

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages29-76
Number of pages48
Volume445
ISBN (Print)9781588298539
DOIs
Publication statusPublished - 2008

Publication series

NameMethods in Molecular Biology
Volume445
ISSN (Print)10643745

Keywords

  • Apoptosis
  • Autophagosomes
  • Endoplasmic reticulum
  • Fluorescence microscopy
  • LC3-GFP
  • Lysosomes
  • Mitochondria
  • Starvation.

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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