| Detection of autophagy in cell death. | |
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MedLine Citation:
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PMID: 18662576 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Activation of autophagosomes in cell death has been described since the late 1950s as a form of cell death characterized by consumption of the bulk of the cytoplasm by lysosomal derivatives. However, it is not yet established that autophagy is a primary, causative mechanism of death rather than a response to initial problems. Methods to assess autophagic cell death are similar to those used to detect and measure autophagy, with further evidence that the affected cells are indeed dying. These methods include structural analysis using electron microscopy, examination of the activity of lysosomal enzymes, assessment of the number, size, and location of lysosomes by the uptake of fluorescent molecules; measurement of the activity of autophagy-related genes such as the cleavage and activation of LC3 by Western blotting or green fluorescent protein tagging; evaluation of the effects of inhibition of one or more lysosomal enzymes, inhibition of fusion of organelles, or inhibition of intercompartmental transfer of molecules by putatively specific inhibitors; and interference with cells in order to change the expression of components of the lysosomal system to study the effect of this change on autophagy and autophagic cell death. |
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Authors:
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Zahra Zakeri; Alicia Melendez; Richard A Lockshin |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Methods in enzymology Volume: 442 ISSN: 1557-7988 ISO Abbreviation: Meth. Enzymol. Publication Date: 2008 |
Date Detail:
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Created Date: 2008-07-29 Completed Date: 2009-01-09 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0212271 Medline TA: Methods Enzymol Country: United States |
Other Details:
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Languages: eng Pagination: 289-306 Citation Subset: IM |
Affiliation:
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Department of Biology, Queens College and Graduate Center of the City University of New York, Flushing, New York, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Apoptosis / physiology* Autophagy / physiology* Blotting, Western Cell Death / physiology Green Fluorescent Proteins / genetics, metabolism Humans Lysosomes / metabolism*, ultrastructure Microscopy, Confocal Microscopy, Electron, Transmission Microscopy, Fluorescence Phagosomes / metabolism, ultrastructure Recombinant Fusion Proteins / genetics, metabolism |
| Chemical | |
Reg. No./Substance:
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0/Recombinant Fusion Proteins; 147336-22-9/Green Fluorescent Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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