Comparison of two methods of staining apoptotic cells of leukemia cell lines: Terminal deoxynucleotidyl transferase and DNA polymerase I reactions

Ichiro Yamadori, Tadashi Yoshino, Eisaku Kondo, Liu Cao, Tadaatsu Akagi, Yoshinobu Matsuo, Jun Minowada

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

We compared two methods to stain apoptotic cells, one using terminal deoxynucleotidyl transferase (TDT), the other DNA polymerase I, using leukemia cell lines treated with anti-Fas monoclonal antibody (MAb). Both TDT and polymerase I strongly reacted with fragmented nuclei of apoptotic MOLT- 16 and Jurkat cells, but only polymerase I strongly reacted with nonfragmented nuclei of early apoptotic cells. Anti-Fas MAb-treated MOLT-4 cells showed morphological changes corresponding to early apoptosis and were strongly positive for polymerase I only. MOLT-16 and Jurkat cells treated with anti-Fas MAb and inhibitors of endonuclease and poly(ADP-ribose) polymerase showed the morphology of early apoptosis but were not strongly stained by TDT. Because DNA polymerase I has nick-translation activity, it is possible that DNA polymerase I reaction is positive in early apoptotic cells by detecting single-strand DNA cleavage, which occurs before extensive oligonucleosomal DNA cleavage and late morphological changes of apoptosis in leukemia cell lines. Although TDT is widely used to stain apoptotic cells, DNA polymerase I may be more applicable in special cases of apoptosis, in which cells undergo single-strand rather than double-strand DNA breaks. However, the procedure has limitations, such as the necessity to use cell smears for comparison with the TDT reaction.

Original languageEnglish
Pages (from-to)85-90
Number of pages6
JournalJournal of Histochemistry and Cytochemistry
Volume46
Issue number1
Publication statusPublished - Jan 1998

Fingerprint

DNA Polymerase I
DNA Nucleotidylexotransferase
anti-Fas monoclonal antibody
Leukemia
Staining and Labeling
Cell Line
Apoptosis
DNA Cleavage
Jurkat Cells
Coloring Agents
Double-Stranded DNA Breaks
Endonucleases

Keywords

  • Apoptosis
  • DNA cleavage
  • DNA polymerase I
  • Leukemia cell line
  • Terminal deoxynucleotidyl transferase

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

Cite this

Comparison of two methods of staining apoptotic cells of leukemia cell lines : Terminal deoxynucleotidyl transferase and DNA polymerase I reactions. / Yamadori, Ichiro; Yoshino, Tadashi; Kondo, Eisaku; Cao, Liu; Akagi, Tadaatsu; Matsuo, Yoshinobu; Minowada, Jun.

In: Journal of Histochemistry and Cytochemistry, Vol. 46, No. 1, 01.1998, p. 85-90.

Research output: Contribution to journalArticle

Yamadori, Ichiro ; Yoshino, Tadashi ; Kondo, Eisaku ; Cao, Liu ; Akagi, Tadaatsu ; Matsuo, Yoshinobu ; Minowada, Jun. / Comparison of two methods of staining apoptotic cells of leukemia cell lines : Terminal deoxynucleotidyl transferase and DNA polymerase I reactions. In: Journal of Histochemistry and Cytochemistry. 1998 ; Vol. 46, No. 1. pp. 85-90.
@article{a90e09a098ba4ef1af674f2ba0f39d21,
title = "Comparison of two methods of staining apoptotic cells of leukemia cell lines: Terminal deoxynucleotidyl transferase and DNA polymerase I reactions",
abstract = "We compared two methods to stain apoptotic cells, one using terminal deoxynucleotidyl transferase (TDT), the other DNA polymerase I, using leukemia cell lines treated with anti-Fas monoclonal antibody (MAb). Both TDT and polymerase I strongly reacted with fragmented nuclei of apoptotic MOLT- 16 and Jurkat cells, but only polymerase I strongly reacted with nonfragmented nuclei of early apoptotic cells. Anti-Fas MAb-treated MOLT-4 cells showed morphological changes corresponding to early apoptosis and were strongly positive for polymerase I only. MOLT-16 and Jurkat cells treated with anti-Fas MAb and inhibitors of endonuclease and poly(ADP-ribose) polymerase showed the morphology of early apoptosis but were not strongly stained by TDT. Because DNA polymerase I has nick-translation activity, it is possible that DNA polymerase I reaction is positive in early apoptotic cells by detecting single-strand DNA cleavage, which occurs before extensive oligonucleosomal DNA cleavage and late morphological changes of apoptosis in leukemia cell lines. Although TDT is widely used to stain apoptotic cells, DNA polymerase I may be more applicable in special cases of apoptosis, in which cells undergo single-strand rather than double-strand DNA breaks. However, the procedure has limitations, such as the necessity to use cell smears for comparison with the TDT reaction.",
keywords = "Apoptosis, DNA cleavage, DNA polymerase I, Leukemia cell line, Terminal deoxynucleotidyl transferase",
author = "Ichiro Yamadori and Tadashi Yoshino and Eisaku Kondo and Liu Cao and Tadaatsu Akagi and Yoshinobu Matsuo and Jun Minowada",
year = "1998",
month = "1",
language = "English",
volume = "46",
pages = "85--90",
journal = "Journal of Histochemistry and Cytochemistry",
issn = "0022-1554",
publisher = "Histochemical Society Inc.",
number = "1",

}

TY - JOUR

T1 - Comparison of two methods of staining apoptotic cells of leukemia cell lines

T2 - Terminal deoxynucleotidyl transferase and DNA polymerase I reactions

AU - Yamadori, Ichiro

AU - Yoshino, Tadashi

AU - Kondo, Eisaku

AU - Cao, Liu

AU - Akagi, Tadaatsu

AU - Matsuo, Yoshinobu

AU - Minowada, Jun

PY - 1998/1

Y1 - 1998/1

N2 - We compared two methods to stain apoptotic cells, one using terminal deoxynucleotidyl transferase (TDT), the other DNA polymerase I, using leukemia cell lines treated with anti-Fas monoclonal antibody (MAb). Both TDT and polymerase I strongly reacted with fragmented nuclei of apoptotic MOLT- 16 and Jurkat cells, but only polymerase I strongly reacted with nonfragmented nuclei of early apoptotic cells. Anti-Fas MAb-treated MOLT-4 cells showed morphological changes corresponding to early apoptosis and were strongly positive for polymerase I only. MOLT-16 and Jurkat cells treated with anti-Fas MAb and inhibitors of endonuclease and poly(ADP-ribose) polymerase showed the morphology of early apoptosis but were not strongly stained by TDT. Because DNA polymerase I has nick-translation activity, it is possible that DNA polymerase I reaction is positive in early apoptotic cells by detecting single-strand DNA cleavage, which occurs before extensive oligonucleosomal DNA cleavage and late morphological changes of apoptosis in leukemia cell lines. Although TDT is widely used to stain apoptotic cells, DNA polymerase I may be more applicable in special cases of apoptosis, in which cells undergo single-strand rather than double-strand DNA breaks. However, the procedure has limitations, such as the necessity to use cell smears for comparison with the TDT reaction.

AB - We compared two methods to stain apoptotic cells, one using terminal deoxynucleotidyl transferase (TDT), the other DNA polymerase I, using leukemia cell lines treated with anti-Fas monoclonal antibody (MAb). Both TDT and polymerase I strongly reacted with fragmented nuclei of apoptotic MOLT- 16 and Jurkat cells, but only polymerase I strongly reacted with nonfragmented nuclei of early apoptotic cells. Anti-Fas MAb-treated MOLT-4 cells showed morphological changes corresponding to early apoptosis and were strongly positive for polymerase I only. MOLT-16 and Jurkat cells treated with anti-Fas MAb and inhibitors of endonuclease and poly(ADP-ribose) polymerase showed the morphology of early apoptosis but were not strongly stained by TDT. Because DNA polymerase I has nick-translation activity, it is possible that DNA polymerase I reaction is positive in early apoptotic cells by detecting single-strand DNA cleavage, which occurs before extensive oligonucleosomal DNA cleavage and late morphological changes of apoptosis in leukemia cell lines. Although TDT is widely used to stain apoptotic cells, DNA polymerase I may be more applicable in special cases of apoptosis, in which cells undergo single-strand rather than double-strand DNA breaks. However, the procedure has limitations, such as the necessity to use cell smears for comparison with the TDT reaction.

KW - Apoptosis

KW - DNA cleavage

KW - DNA polymerase I

KW - Leukemia cell line

KW - Terminal deoxynucleotidyl transferase

UR - http://www.scopus.com/inward/record.url?scp=0031965272&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031965272&partnerID=8YFLogxK

M3 - Article

C2 - 9405497

AN - SCOPUS:0031965272

VL - 46

SP - 85

EP - 90

JO - Journal of Histochemistry and Cytochemistry

JF - Journal of Histochemistry and Cytochemistry

SN - 0022-1554

IS - 1

ER -