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In situ detection of fragmented dna (tunel assay) fails to discriminate among apoptosis, necrosis, and autolytic cell death: A cautionary note
Author(s) -
Kraupp Bettina Grasl,
RuttkayNedecky Branislav,
Koudelka Helga,
Bukowska Krystyna,
Bursch Wilfried,
SchulteHermann Rolf
Publication year - 1995
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.1840210534
Subject(s) - tunel assay , dna fragmentation , apoptosis , apoptotic dna fragmentation , biology , programmed cell death , fragmentation (computing) , microbiology and biotechnology , necrosis , terminal deoxynucleotidyl transferase , in situ nick end labeling , pathology , biochemistry , medicine , genetics , ecology
Detection of DNA fragments in situ using the terminal deoxyribonucleotidyl transferase (TDT)‐mediated dUTP‐digoxigenin nick end labeling (TUNEL) assay is increasingly applied to investigate active cell death (apoptosis). We studied the specificity of the assay in well‐defined models of apoptosis and necrosis as well as in postmortem autolysis in rat liver. During involution of liver hyperplasia, which follows stopping treatment with the hepatomitogens cyproterone acetate (CPA) or nafenopin (NAF), numerous apoptotic hepatocytes could be observed with TUNEL‐positive chromatin residues. A similar TUNEL‐positive reaction appeared in necrotic hepatocytes after a cytotoxic dose of carbon tetrachloride (CCl 4 ) or N ‐nitrosomorpholine (NNM). Also, in insufficiently fixed, autolytic livers TUNEL‐positive nuclei were observed. Thus, DNA fragmentation is common to different kinds of cell death; its detection in situ should not be considered a specific marker of apoptosis.