Early detection of staurosporine-induced apoptosis by comet and annexin V assays

Comet, TUNEL, and annexin V assays were used to identify DNA fragmentation and plasma membrane alterations occurring during staurosporine-induced apoptosis in Chinese hamster ovary cells. TUNEL assay detected apoptotic cells after 6 h treatment. The occurrence of annexin V immunofluorescence stainin...

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Bibliographic Details
Published in:Histochemistry and cell biology 1999-08, Vol.112 (2), p.155-161
Main Authors: Godard, T, Deslandes, E, Lebailly, P, Vigreux, C, Sichel, F, Poul, J M, Gauduchon, P
Format: Article
Language:English
Subjects:
Animals
Annexin A5 - analysis
Annexin V
Apoptosis
CHO Cells
Comet assay
Cricetinae
Deoxyribonucleic acid
DNA
DNA Fragmentation
Immunofluorescence
In Situ Nick-End Labeling
Phosphatidylserine
Phosphatidylserines - metabolism
Staurosporine
Staurosporine - pharmacology
Time Factors
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Summary:Comet, TUNEL, and annexin V assays were used to identify DNA fragmentation and plasma membrane alterations occurring during staurosporine-induced apoptosis in Chinese hamster ovary cells. TUNEL assay detected apoptotic cells after 6 h treatment. The occurrence of annexin V immunofluorescence staining after 1 h treatment confirms that exposure of phosphatidylserine (PS) residues is an early biochemical feature of apoptosis. According to intensity, three annexin staining patterns were distinguished, related to different steps in the apoptotic process. The detection of highly damaged cells by the comet assay after 3 h treatment occurred earlier than the detection of DNA modifications by the TUNEL assay, but later than the exposure of PS residues. However, late apoptotic cells, otherwise characterized by plasma membrane disruption and high annexin V staining, were not detected by the comet assay. In this case, comet assay modified by omitting electrophoresis (halo assay) was more sensitive for an accurate quantification of the apoptotic fraction.
ISSN:0948-6143
1432-119X
DOI:10.1007/s004180050402