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

Citation
Jk. Cui et al., Oxidative DNA damage precedes DNA fragmentation after experimental stroke in rat brain, FASEB J, 14(7), 2000, pp. 955-967
Citations number
65
Categorie Soggetti
Experimental Biology
Journal title
FASEB JOURNAL
ISSN journal
08926638 → ACNP
Volume
14
Issue
7
Year of publication
2000
Pages
955 - 967
Database
ISI
SICI code
0892-6638(200005)14:7<955:ODDPDF>2.0.ZU;2-C
Abstract
Experimental stroke using a focal cerebral ischemia and reperfusion (FCIR) model was induced in male Long Evans rats by a bilateral occlusion of both common carotid arteries and the right middle cerebral artery for 30-90 min, followed by various periods of reperfusion, Oxidative DNA lesions in the i psilateral cortex were demonstrated using Escherichia coli formamidopyrimid ine DNA N-glycosylase (Fpg protein)-sensitive sites (FPGSS), as labeled in situ using digoxigenin-dUTP and detected using antibodies against digoxigen in. Because Fpg protein removes 8-hydroxy-2'-deoxyguanine (oh8dG) and other lesions in DNA, FPGSS measure oxidative DNA damage. The number of FPGSS-po sitive cells in the cortex from the sham-operated control group was 3 +/- 3 (mean +/- SD per mm(2)), In animals that received 90 min occlusion and 15 min of reperfusion (FCIR 90/15), FPGSS-positive cells were significantly in creased by 200-fold. Oxidative DNA damage was confirmed by using monoclonal antibodies against 8-hydroxy-guanosine (oh8G) and oh8dG, A pretreatment of RNase A (100 mu g/ml) to the tissue reduced, but did not abolish, the oh8d G signal, The number of animals with positive FPGSS or oh8dG was significan tly (P<0.01) higher in the FCIR group than in the sham-operated control gro up. We detected few FPGSS of oh8dG-positive cells in the animals treated wi th FCIR of 90/60, No terminal UTP nicked-end labeling (TUNEL)-positive cell s, as a detection of cell death, were detected at this early reperfusion ti me. Our data suggest that early oxidative DNA lesions elicited by experimen tal stroke could be repaired. Therefore, the oxidative DNA lesions observed in the nuclear and mitochondrial DNA of the brain are different from the D NA fragmentation detected using TUNEL.