Oxidative Cellular Damage and the Reduction of APE/Ref-1 Expression after Experimental Traumatic Brain Injury

doi:10.1006/nbdi.2001.0396

Neurobiology of Disease

Volume 8, Issue 3, June 2001, Pages 380-390

https://doi.org/10.1006/nbdi.2001.0396 Get rights and content

Abstract

The DNA repair enzyme, apurinic/apyrimidinic endonuclease (or redox effector factor-1, APE/Ref-1), is involved in base excision repair of apurinic/apyrimidinic sites after oxidative DNA damage. We investigated the expression of APE/Ref-1 and its relationship to oxidative stress after severe traumatic brain injury produced by controlled cortical impact in normal mice, and in mice over- or underexpressing copper-zinc superoxide dismutase (SOD1TG and SOD1KO, respectively). Oxygen free radical-mediated cellular injury was visualized with 8-hydroxyguanine immunoreactivity as a marker for DNA oxidation, and in situ hydroethidine oxidation as a marker for superoxide production. After trauma there was a reduced expression of APE/Ref-1 in the ipsilateral cortex and hippocampus that correlated with the gene dosage levels of cytosolic superoxide dismutase. The decrease in APE/Ref-1 expression preceded DNA fragmentation. There was also a close correlation between APE/Ref-1 protein levels 4 h after trauma and the volume of the lesion 1 week after injury. Our data have demonstrated that reduction of APE/Ref-1 protein levels correlates closely with the level of oxidative stress after traumatic brain injury. We suggest that APE/Ref-1 immunoreactivity is a sensitive marker for oxidative cellular injury.

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The resulting mitochondrial dysfunction also promotes ROS formation. In addition, lipid peroxidation, infiltrated macrophages/neutrophils, induction of NADPH oxidases NOX2 and NOX4, and activation of neuronal nitric oxide synthase after TBI leads to increased formation of ROS including superoxide radical, hydroxyl radical, and non-radicals like hydrogen peroxide and singlet oxygen and reactive nitrogen species like nitric oxide (Lewén et al., 2001; Ma et al., 2018). All these molecules lead to extensive DNA damage (Halliwell and Gutteridge, 2015).
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In some selected regions of the central nervous system, APE1 is highly expressed [141,142], and reduction of its expression leads to an increase in the rate of apoptosis. This process occurs in the cerebral cortex following compression injury [143], in the hippocampus after hypoxic-ischemic insult [144] and in the spinal cord after ischemia [145]. Increased expression of APE1 in the hippocampus was observed in patients with Alzheimer's disease [146].
Protein moonlighting is a phenomenon in which a single polypeptide chain can perform a number of different unrelated functions. Here we present our analysis of moonlighting in the case of selected DNA repair proteins which include G:T mismatch-specific thymine DNA glycosylase (TDG), methyl-CpG-binding domain protein 4 (MBD4), apurinic/apyrimidinic endonuclease 1 (APE1), AlkB homologs, poly (ADP-ribose) polymerase 1 (PARP-1) and single-strand selective monofunctional uracil DNA glycosylase 1 (SMUG1). Most of their additional functions are not accidental and clear patterns are emerging. Participation in RNA metabolism is not surprising as bases occurring in RNA are the same or very similar to those in DNA. Other common additional function involves regulation of transcription. This is not unexpected as these proteins bind to specific DNA regions for DNA repair, hence they can also be recruited to regulate transcription. Participation in demethylation and replication of DNA appears logical as well. Some of the multifunctional DNA repair proteins play major roles in many diseases, including cancer. However, their moonlighting might prove a major difficulty in the development of new therapies because it will not be trivial to target a single protein function without affecting its other functions that are not related to the disease.
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With increased survival rates for cancer patients, toxicity secondary to anticancer therapies has become a prevalent clinical concern. Of major importance are toxicities occurring in postmitotic cells (cardiac muscle, skeletal muscle, and neurons) since these toxicities are quite debilitating, they often persist long after therapy is discontinued, and few interventions exist to prevent or reverse them. Although the mechanisms mediating these toxicities are not known, one promising area that requires further exploration is the ability of DNA repair mechanisms to reverse the toxic effects of a number of anticancer drugs. For example, studies in animal models show that enhancing the base excision repair pathway attenuates neuronal damage by chemotherapeutic agents, suggesting that manipulating DNA repair mechanisms may be a novel approach to diminish neurotoxicity during or after cancer therapy. Whether DNA damage and repair are critical in cardiac and skeletal muscle toxicity after cancer therapy remains to be determined.
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Expression was high throughout the organ during embryonic development, but reduced in all regions post natal except the hippocampus, where mRNA levels remained high in the pyramidal and granule cells at post natal day 21. This early finding was supported by subsequent studies (Edwards et al., 1998a; Gillardon et al., 1997; Lewen et al., 2001; Walton et al., 1997) more focused on characterizing changes in APE1 protein levels after oxidative damage, while offering further insight into protein localization (information collated in Table 2). To summarize these studies, APE1 in the hippocampus can be induced after moderate DNA damage; this protective response is transient and does not occur for longer than 24–36 h.
The repair of damaged DNA is essential to maintain longevity of an organism. The brain is a matrix of different neural cell types including proliferative astrocytes and post-mitotic neurons. Post-mitotic DNA repair is a version of proliferative DNA repair, with a reduced number of available pathways and most of these attenuated. Base excision repair (BER) is one pathway that remains robust in neurons; it is this pathway that resolves the damage due to oxidative stress. This oxidative damage is an unavoidable byproduct of respiration, and considering the high metabolic activity of neurons this type of damage is particularly pertinent in the brain. The accumulation of oxidative DNA damage over time is a central aspect of the theory of aging and repair of such chronic damage is of the highest importance. We review research conducted in BER mouse models to clarify the role of this pathway in the neural system. The requirement for BER in proliferating cells also correlates with high levels of many of the BER enzymes in neurogenesis after DNA damage. However, the pathway is also necessary for normal neural maintenance as larger infarct volumes after ischemic stroke are seen in some glycosylase deficient animals. Further, the requirement for DNA polymerase β in post-mitotic BER is potentially more important than in proliferating cells due to reduced levels of replicative polymerases. The BER response may have particular relevance for the onset and progression of many neurodegenerative diseases associated with an increase in oxidative stress including Alzheimer's.
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Oxidative stress has been identified as an important contributor to neurodegeneration associated with acute CNS injuries and diseases such as spinal cord injury (SCI), traumatic brain injury (TBI), and ischemic stroke. In this review, we briefly detail the damaging effects of oxidative stress (lipid peroxidation, protein oxidation, etc.) with a particular emphasis on DNA damage. Evidence for DNA damage in acute CNS injuries is presented along with its downstream effects on neuronal viability. In particular, unchecked oxidative DNA damage initiates a series of signaling events (e.g. activation of p53 and PARP-1, cell cycle re-activation) which have been shown to promote neuronal loss following CNS injury. These findings suggest that preventing DNA damage might be an effective way to promote neuronal survival and enhance neurological recovery in these conditions. Finally, we identify the telomere and telomere-associated proteins (e.g. telomerase) as novel therapeutic targets in the treatment of neurodegeneration due to their ability to modulate the neuronal response to both oxidative stress and DNA damage.

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Regular ArticleOxidative Cellular Damage and the Reduction of APE/Ref-1 Expression after Experimental Traumatic Brain Injury

Abstract

Free Rad. Biol. Med.

Arch. Biochem. Biophys.

Free Rad. Biol. Med.

Mutat. Res.

Brain Res.

Mutat. Res.

Methods Enzymol.

Neurosci. Lett.

Neuroscience

J. Biol. Chem.

Free Rad. Biol. Med.

Am. J. Pathol.

The use of salicylate hydroxylation to detect hydroxyl radical generation in ischemic and traumatic brain injury. Reversal by tirilazad mesylate (U-74006F)

Mol. Chem. Neuropathol.

Interaction of human apurinic endonuclease and DNA polymerase beta in the base excision repair pathway

Proc. Natl. Acad. Sci. USA

Superoxide production in rat hippocampal neurons: Selective imaging with hydroethidine

J. Neurosci.

Human apurinic/apyrimidinic endonuclease is processive

Biochemistry

Early detection of DNA strand breaks in the brain after transient focal ischemia: Implications for the role of DNA damage in apoptosis and neuronal cell death

J. Neurochem.

DNA damage and repair in central nervous system injury: National Institute of Neurological Disorders and Stroke workshop summary

Stroke

Oxidative DNA damage precedes DNA fragmentation after experimental stroke in rat brain

FASEB J.

A controlled cortical impact model of traumatic brain injury in the rat

J. Neurosci. Methods.

Mechanism of action of a mammalian DNA repair endonuclease

Biochemistry

APE/Ref-1 responses to ischemia in rat brain

Neuroreport

Transgenic mice with increased Cu/Zn-superoxide dismutase activity: Animal model of dosage effects in Down syndrome

Proc. Natl. Acad. Sci. USA

The 21-aminosteroid U-74389G reduces cerebral superoxide anion concentration following fluid percussion injury of the brain

J. Neurotrauma

Early decrease of apurinic/apyrimidinic endonuclease expression after transient focal cerebral ischemia in mice

J. Cereb. Blood Flow Metab.

Regular Article
Oxidative Cellular Damage and the Reduction of APE/Ref-1 Expression after Experimental Traumatic Brain Injury