Histochemical Demonstration of DNA Double Strand Breaks by in Situ 3′-tailing Reaction in Apoptotic Endometrium
作者:
NakamuraToshitsugu,
SakaiToshihiko,
HotchiMasao,
期刊:
Biotechnic&Histochemistry
(Taylor Available online 1995)
卷期:
Volume 70,
issue 1
页码: 33-39
ISSN:1052-0295
年代: 1995
DOI:10.3109/10520299509108314
出版商: Taylor&Francis
关键词: DNA double strand breaks;apoptosis;3′-tailing reaction;terminal deoxynucleotidyl transferase;λ-exonuclease;histochemistry
数据来源: Taylor
摘要:
A new histochemical technique, called in situ 3′-tailing reaction (ISTR), to detect DNA double strand breaks (DSB) was developed and applied to tissue sections of apoptotic endometrium. To demonstrate DSB, biotin-labeled and unlabeled dATPs with terminal deoxynucleotidyl transferase (TdT) were added to the many 3-hydroxyl termini of DNA fragments generated in the apoptotic cells. For an efficient 3′-end labeling, it was necessary to treat the sections withλ-exonuclease (λEx) prior to the TdT reaction to generate 3′-protruding ends. TheλEx-TdT reaction specifically labeled nuclear fragments in the apoptotic cells in paraformaldehyde fixed frozen sections. In paraffin sections, pretreatment with proteinase K was effective for 3′-tailing reaction. ISTR should be a useful tool for detecting dying cells in both physiological and pathological states.
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