Propidium iodide staining correlates with the extent of DNA degradation in isolated nuclei

Details

Serval ID
serval:BIB_9037B5DE95A7
Type
Article: article from journal or magazin.
Collection
Publications
Title
Propidium iodide staining correlates with the extent of DNA degradation in isolated nuclei
Journal
Biochemical and Biophysical Research Communications
Author(s)
Crompton  T., Peitsch  M. C., MacDonald  H. R., Tschopp  J.
ISSN
0006-291X (Print)
Publication state
Published
Issued date
03/1992
Volume
183
Number
2
Pages
532-7
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Mar 16
Abstract
Gradual degradation of internucleosomal DNA is a hallmark of apoptosis and can be simulated by incubating isolated thymocyte nuclei in the presence of 5 mM Mg2+ and 5 mM Ca2+ at 37 degrees C. Staining of nuclei with the DNA binding fluorescent dye propidium iodide (PI) showed that intensity of fluorescence correlated with the extent of DNA degradation. PI fluorescence was increased in the presence of DNase I. Thus it seems that the cleavage of chromatin DNA by DNase 1 or by the endogenous enzyme increases the accessibility of DNA for the dye. No increase of fluorescence was observed in the presence of the known inhibitors of the endogenous endonuclease: Zn2+ and EGTA. However, the presence of Zn2+ led to decreased staining of the nuclei by PI and caused a shift in the scatter profile of the nuclei, suggesting that a conformational change of chromatin is induced by this ion. This correlation between intensity of PI staining and DNA degradation should be useful to compare endogenous nuclease levels in lymphocyte populations.
Keywords
Animals Cations/pharmacology Cell Nucleus/chemistry DNA/*chemistry *DNA Damage Flow Cytometry/*methods Mice Mice, Inbred BALB C *Propidium *Staining and Labeling Thymus Gland/cytology
Pubmed
Web of science
Create date
24/01/2008 16:18
Last modification date
03/03/2018 19:24
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