Caspase-activated DNase/DNA fragmentation factor 40 mediates apoptotic DNA fragmentation in transient cerebral ischemia and in neuronal cultures

Guodong Cao, Wei Pei, Jing Lan, R. Anne Stetler, Yumin Luo, Tetsuya Nagayama, Steven H. Graham, Xiao Ming Yin, Roger P. Simon, Jun Chen

Research output: Contribution to journalArticle

86 Scopus citations

Abstract

Nuclear changes, including internucleosomal DNA fragmentation, are characteristic features of neuronal apoptosis resulting from transient cerebral ischemia and related brain insults for which the molecular mechanism has not been elucidated. Recent studies suggest that a caspase-3-mediated mechanism may be involved in the process of nuclear degradation in ischemic neurons. In this study, we cloned from rat brain a homolog cDNA encoding caspase-activated deoxyribonuclease (CAD)/ DNA fragmentation factor 40 (DFF40), a 40 kDa nuclear enzyme that is activated by caspase-3 and promotes apoptotic DNA degradation. Subsequently, we investigated the role of CAD/ DFF40 in the induction of internucleosomal DNA fragmentation in the hippocampus in a rat model of transient global ischemia and in primary neuronal cultures under ischemia-like conditions. At 8-72 hr after ischemia, CAD/DFF40 mRNA and protein were induced in the degenerating hippocampal CA1 neurons. CAD/DFF40 formed a heterodimeric complex in the nucleus with its natural inhibitor CAD (ICAD) and was activated after ischemia in a delayed manner (>24 hr) by caspase-3, which translocated into the nucleus and cleaved ICAD. Furthermore, an induced CAD/DFF40 activity was detected in nuclear extracts in both in vivo and in vitro models, and the DNA degradation activity of CAD/DFF40 was inhibited by purified ICAD protein. These results strongly suggest that CAD/DFF40 is the endogenous endonuclease that mediates caspase-3-dependent internucleosomal DNA degradation and related nuclear alterations in ischemic neurons.

Original languageEnglish (US)
Pages (from-to)4678-4690
Number of pages13
JournalJournal of Neuroscience
Volume21
Issue number13
DOIs
StatePublished - Jul 1 2001

Keywords

  • Apoptosis
  • Caspase-3
  • Caspase-activated deoxyribonuclease
  • Cerebral ischemia
  • DNA fragmentation
  • Programmed cell death

ASJC Scopus subject areas

  • Neuroscience(all)

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    Cao, G., Pei, W., Lan, J., Stetler, R. A., Luo, Y., Nagayama, T., Graham, S. H., Yin, X. M., Simon, R. P., & Chen, J. (2001). Caspase-activated DNase/DNA fragmentation factor 40 mediates apoptotic DNA fragmentation in transient cerebral ischemia and in neuronal cultures. Journal of Neuroscience, 21(13), 4678-4690. https://doi.org/10.1523/jneurosci.21-13-04678.2001