On the use of an appropriate TdT-mediated dUTP-biotin nick end labeling assay to identify apoptotic cells.

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State: Public
Version: Author's accepted manuscript
Serval ID
serval:BIB_A460A6316451
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
On the use of an appropriate TdT-mediated dUTP-biotin nick end labeling assay to identify apoptotic cells.
Journal
Analytical Biochemistry
Author(s)
Lebon C., Rodriguez G.V., Zaoui I.E., Jaadane I., Behar-Cohen F., Torriglia A.
ISSN
1096-0309 (Electronic)
ISSN-L
0003-2697
Publication state
Published
Issued date
2015
Peer-reviewed
Oui
Volume
480
Pages
37-41
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov'tPublication Status: ppublish
Abstract
Apoptosis is an essential cellular mechanism involved in many processes such as embryogenesis, metamorphosis, and tissue homeostasis. DNA fragmentation is one of the key markers of this form of cell death. DNA fragmentation is executed by endogenous endonucleases such as caspase-activated DNase (CAD) in caspase-dependent apoptosis. The TUNEL (TdT-mediated dUTP-biotin nick end labeling) technique is the most widely used method to identify apoptotic cells in a tissue or culture and to assess drug toxicity. It is based on the detection of 3'-OH termini that are labeled with dUTP by the terminal deoxynucleotidyl transferase. Although the test is very reliable and sensitive in caspase-dependent apoptosis, it is completely useless when cell death is mediated by pathways involving DNA degradation that generates 3'-P ends as in the LEI/L-DNase II pathway. Here, we propose a modification in the TUNEL protocol consisting of a dephosphorylation step prior to the TUNEL labeling. This allows the detection of both types of DNA breaks induced during apoptosis caspase-dependent and independent pathways, avoiding underestimating the cell death induced by the treatment of interest.
Pubmed
Web of science
Open Access
Yes
Create date
21/04/2015 10:01
Last modification date
20/08/2019 15:09
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