Inhibitory effect of intracellular lipid load on macroautophagy

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Degradation of intracellular components via macroautophagy is a complex multistep process that starts with the sequestration of cytosolic cargo in a de novo formed double-membrane vesicle or autophagosome. This compartment acquires the hydrolases required for cargo digestion by fusion with lysosomes. In contrast to the detailed molecular dissection of the components that participate in the induction, regulation and execution of the early steps in macroautophagy through the engulfment of cargo in autophagosomes, the mechanisms involved in the lysosomal clearance of autophagosomes have been poorly characterized in mammals. One of the major limitations in this respect has been the fact that autophagosome-lysosome fusion in intact cells involves several independent steps, namely binding of the molecular motors associated with the surface of the vesicles with the cytoskeletal network, directional vesicular trafficking and fusion between the two vesicular compartments. Furthermore, both lysosomes and autophagosomes are very dynamic organelles that can fuse with different vesicular structures involved in macroautophagy, but also along the endocytic and phagocytic pathways. To resolve these limitations and directly analyze the fusion step between autophagosomes and different compartments of the endocytic-lysosomal pathway, we have recently developed an in vitro fusion assay with autophagosomes, lysosomes and endosomes isolated from cells or tissues. Fluorescent labeling of these compartments allows for the tracking of fusion events by fluorescence microscopy or by fluorescence-activated cell sorting (FACS). Labeling of either membrane proteins on the surface of the organelles or dye-loading of the vesicles permits the monitoring of hemi-membrane fusion and complete vesicular fusion (cargo mixing).

Original languageEnglish (US)
Pages (from-to)825-827
Number of pages3
JournalAutophagy
Volume6
Issue number6
DOIs
StatePublished - Aug 16 2010

Fingerprint

Autophagy
Lipids
Lysosomes
Organelles
Membrane Fusion
Endosomes
Hydrolases
Autophagosomes
Fluorescence Microscopy
Dissection
Digestion
Mammals
Flow Cytometry
Membrane Proteins
Coloring Agents
Membranes

Keywords

  • Autophagy
  • Cholesterol
  • Lysosomes
  • Vesicular fusion

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Inhibitory effect of intracellular lipid load on macroautophagy. / Koga, Hiroshi; Kaushik, Susmita; Cuervo, Ana Maria.

In: Autophagy, Vol. 6, No. 6, 16.08.2010, p. 825-827.

Research output: Contribution to journalArticle

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