Differences in ethidium bromide and 4'-6-diamidino-2-phenylindole staining profiles with regard to DNA fragmentation during apoptosis.
To simply and directly evaluate DNA fragmentation during apoptosis induced in mouse cultured hepatocytes by an anti-Fas antibody, we examined the fluorescence intensity in cell nuclei stained with ethidium bromide and 4'-6-diamidino-2-phenylindole by optiphoto fluorescence microscopy. The intensity of the former staining for the nuclear DNA of apoptotic cells was clearly decreased compared to that of non-apoptotic cells, whereas no difference in the fluorescence intensity for the latter stain between the apoptotic and non-apoptotic groups was observed. Thus, the use of optiphoto fluorescence microscopy, in conjunction with both stains, constitutes a useful tool for the evaluation of apoptotic DNA fragmentation.
Shono
M
M
General Laboratory for Medical Research, School of Medicine, University of Tokushima, Japan.
Shimizu
I
I
Omoya
T
T
Hiasa
A
A
Honda
H
H
Tomita
Y
Y
Ito
S
S
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D017209:Apoptosis; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism*; D053938:DNA Fragmentation / Q000187:drug effects; D003609:Dactinomycin / Q000494:pharmacology; D004996:Ethidium / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism*; D007211:Indoles / Q000378:metabolism*; D007364:Intercalating Agents / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000187:drug effects; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D013194:Staining and Labeling; D019014:fas Receptor / Q000276:immunology
D000906:Antibodies; D005456:Fluorescent Dyes; D007211:Indoles; D007364:Intercalating Agents; D019014:fas Receptor; D003609:Dactinomycin; C007293:DAPI; D004247:DNA; D004996:Ethidium