Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Expression of DNase gamma during Fas-independent apoptotic DNA fragmentation in rodent hepatocytes


Endonuclease-induced DNA fragmentation is a hallmark of apoptosis. DNase gamma (DNase γ) was recently identified as one of the endonucleases responsible for apoptotic DNA fragmentation. In this study, immunohistochemistry for DNase γ was performed on paraffin sections of rodent liver in well-defined models of hepatocyte apoptosis induced by Fas antibody (Fas) or cycloheximide (CHX), and necrosis induced by lipopolysaccharide (LPS) or carbon tetrachloride (CCl4). DNase γ immunoreactivity was compared with TdT-mediated dUTP nick-end labeling (TUNEL) reactivity. Our results showed TUNEL reactivity in both apoptotic and necrotic hepatocytes. DNase γ immunoreactivity was not detected during LPS-induced or CCl4-induced hepatocyte necrosis. In contrast, it was evident during CHX-induced, but not Fas-induced, apoptotic DNA fragmentation. These findings suggest that DNase γ plays an important role in Fas-independent apoptotic DNA fragmentation in hepatocytes.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3


  1. Blom WM, Bont HJ de, Meijerman I, Mulder GJ, Nagelkerke JF (1999) Prevention of cycloheximide-induced apoptosis in hepatocytes by adenosine and by caspase inhibitors. Biochem Pharmacol 58:1891–1898

  2. Bohlinger I, Leist M, Gantner F, Angermuller S, Tiegs G, Wendel A (1996) DNA fragmentation in mouse organs during endotoxic shock. Am J Pathol 149:1381–1393

  3. Dong Z, Saikumar P, Weinberg JM, Venkatachalam MA (1997) Internucleosomal DNA cleavage triggered by plasma membrane damage during necrotic cell death. Involvement of serine but not cysteine proteases. Am J Pathol 151:1205–1213

  4. Enari M, Sakahira H, Yokoyama H, Okawa K, Iwamatsu A, Nagata S (1998) A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391:43–50

  5. Gavrieli Y, Sherman Y, Ben-Sasson SA (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119:493–501

  6. Grasl-Kraupp B, Ruttkay-Nedecky B, Koudelka H, Bukowska K, Bursch W, Schulte-Hermann R (1995) In situ detection of fragmented DNA (TUNEL assay) fails to discriminate among apoptosis, necrosis, and autolytic cell death: a cautionary note. Hepatology 21:1465–1468

  7. Higami Y, Tanaka K, Tsuchiya T, Shimokawa I (2000) Intravenous injection of cycloheximide induces apoptosis and up-regulates p53 and Fas receptor expression in the rat liver in vivo. Mutat Res 457:105–111

  8. Higami Y, To K, Ohtani H, Masui K, Iwasaki K, Shiokawa D, Tanuma SI, Shimokawa I (2003) Involvement of DNase gamma in apoptotic DNA fragmentation in histiocytic necrotizing lymphadenitis. Virchows Arch 443:170–174

  9. Ledda-Columbano GM, Coni P, Faa G, Manenti G, Columbano A (1992) Rapid induction of apoptosis in rat liver by cycloheximide. Am J Pathol 140:545–549

  10. Mukae N, Enari M, Sakahira H, Fukuda Y, Inazawa J, Toh H, Nagata S (1998) Molecular cloning and characterization of human caspase-activated DNase. Proc Natl Acad Sci USA 95:9123–9128

  11. Ogasawara J, Watanabe-Fukunaga R, Adachi M, Matsuzawa A, Kasugai T, Kitamura Y, Itoh N, Suda T, Nagata S (1993) Lethal effect of the anti-Fas antibody in mice. Nature 364:806–809

  12. Orrego H, Carmichael FJ, Phillips MJ, Kalant H, Khanna J, Israel Y (1976) Protection by propylthiouracil against carbon tetrachloride-induced liver damage. Gastroenterology 71:821–826

  13. Shiokawa D, Tanuma S (1998) Molecular cloning and expression of a cDNA encoding an apoptotic endonuclease DNase gamma. Biochem J 332:713–720

  14. Shiokawa D, Ohyama H, Yamada T, Takahashi K, Tanuma S (1994) Identification of an endonuclease responsible for apoptosis in rat thymocytes. Eur J Biochem 226:23–30

  15. Shiokawa D, Iwamatsu A, Tanuma S (1997) Purification, characterization, and amino acid sequencing of DNase gamma from rat spleen. Arch Biochem Biophys 346:15–20

  16. Shiokawa D, Tanaka M, Kimura T, Hashizume K, Takasawa R, Ohyama H, Fujita K, Yamada T, Tanuma S (2000) Characterization of two DNase gamma-specific monoclonal antibodies and the in situ detection of DNase gamma in the nuclei of apoptotic rat thymocytes. Biochem Biophys Res Commun 275:343–349

  17. Srinivasan A, Roth KA, Sayers RO, Shindler KS, Wong AM, Fritz LC, Tomaselli KJ (1998) In situ immunodetection of activated caspase-3 in apoptotic neurons in the developing nervous system. Cell Death Differ 5:1004–1016

  18. Tanaka K, Higami Y, Tsuchiya T, Shiokawa D, Tanuma S, Ayabe H, Shimokawa I (2004) Aging increases DNase γ, an apoptosis-related endonuclease, in rat liver nuclei: effect of dietary restriction. Exp Gerontol 39:195–202

  19. Ueda N, Walker PD, Hsu SM, Shah SV (1995) Activation of a 15-kDa endonuclease in hypoxia/reoxygenation injury without morphologic features of apoptosis. Proc Natl Acad Sci USA 92:7202–7206

  20. Yang F, Sun X, Beech W, Teter B, Wu S, Sigel J, Vinters HV, Frautschy SA, Cole GM (1998) Antibody to caspase-cleaved actin detects apoptosis in differentiated neuroblastoma and plaque-associated neurons and microglia in Alzheimer’s disease. Am J Pathol 152:379–389

  21. Zhao QL, Kondo T, Noda A, Fujiwara Y (1999) Mitochondrial and intracellular free-calcium regulation of radiation-induced apoptosis in human leukemic cells. Int J Radiat Biol 75:493–504

Download references

Author information

Correspondence to Yoshikazu Higami.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Higami, Y., Tsuchiya, T., To, K. et al. Expression of DNase gamma during Fas-independent apoptotic DNA fragmentation in rodent hepatocytes. Cell Tissue Res 316, 403–407 (2004).

Download citation


  • DNase gamma
  • Hepatocytes
  • Apoptosis
  • Fas
  • Cycloheximide
  • Rat (F344)
  • Mouse (BALB/c)