TY - JOUR
T1 - Increased mitochondrial DNA oxidative damage after transient middle cerebral artery occlusion in mice
AU - Ohtaki, Hirokazu
AU - Takeda, Takaaki
AU - Dohi, Kenji
AU - Yofu, Sachiko
AU - Nakamachi, Tomoya
AU - Satoh, Kazue
AU - Hiraizumi, Yutaka
AU - Miyaoka, Hideyo
AU - Matsunaga, Masaji
AU - Shioda, Seiji
N1 - Funding Information:
The authors wish to thank Roxanne Reger for editorial help. This work was supported in part by a Showa University Grant-in-Aid for Innovative Collaborative Research Projects (H.O and Y.H) and Research on Health Sciences focusing on Drug Innovation from The Japan Health Science Foundation (M.M and S.S).
PY - 2007/8
Y1 - 2007/8
N2 - Oxidative stress and DNA oxidation play important roles in the induction of ischemic neuronal cell death. However, the subcellular source of oxidized DNA detected by 8-hydroxy-2′-deoxyguanosine (8-OHdG) after ischemia has not been clarified although it is known to increase in the brain after ischemia. One-hour transient ischemia of the middle cerebral artery was induced in mice utilizing an intraluminal filament. The occurrence of superoxide anion as an ethidium (Et) signal, 8-OHdG, cytochrome c release and neuronal cell death were examined using immunohistological and biochemical techniques in sham-operated control (0 h) and 1, 3, 6, 24, or 96 h after reperfusion. Et signals were prominent in the cortical neurons of ipsilateral hemisphere 3 h after reperfusion. Strong 8-OHdG immunoreactivity was observed 3-6 h after reperfusion. Immunoassays after cell fractionation revealed a significant increase of 8-OHdG in mitochondria 6 h after reperfusion. Immunohistochemistry revealed that the 8-OHdG immunoreactivity colocalized with a neuronal marker, microfilament 200 and a mitochondrial marker, cytochrome oxidase subunit I. Cytochrome c rose in cytoplasm at 6 h and TUNEL-positive neurons noted 6-24 h after ischemia. The present results suggest the possibility that the mitochondrial damage including mitochondrial DNA oxidation might be responsible for the induction of ischemic neuronal cell death.
AB - Oxidative stress and DNA oxidation play important roles in the induction of ischemic neuronal cell death. However, the subcellular source of oxidized DNA detected by 8-hydroxy-2′-deoxyguanosine (8-OHdG) after ischemia has not been clarified although it is known to increase in the brain after ischemia. One-hour transient ischemia of the middle cerebral artery was induced in mice utilizing an intraluminal filament. The occurrence of superoxide anion as an ethidium (Et) signal, 8-OHdG, cytochrome c release and neuronal cell death were examined using immunohistological and biochemical techniques in sham-operated control (0 h) and 1, 3, 6, 24, or 96 h after reperfusion. Et signals were prominent in the cortical neurons of ipsilateral hemisphere 3 h after reperfusion. Strong 8-OHdG immunoreactivity was observed 3-6 h after reperfusion. Immunoassays after cell fractionation revealed a significant increase of 8-OHdG in mitochondria 6 h after reperfusion. Immunohistochemistry revealed that the 8-OHdG immunoreactivity colocalized with a neuronal marker, microfilament 200 and a mitochondrial marker, cytochrome oxidase subunit I. Cytochrome c rose in cytoplasm at 6 h and TUNEL-positive neurons noted 6-24 h after ischemia. The present results suggest the possibility that the mitochondrial damage including mitochondrial DNA oxidation might be responsible for the induction of ischemic neuronal cell death.
KW - 8-Hydroxy-2′-deoxyguanosine (8-OHdG)
KW - Cytochrome c
KW - DNA oxidative damage
KW - Mitochondria
KW - Neuronal cell death
KW - Transient focal ischemia
UR - http://www.scopus.com/inward/record.url?scp=34547607954&partnerID=8YFLogxK
U2 - 10.1016/j.neures.2007.04.005
DO - 10.1016/j.neures.2007.04.005
M3 - 学術論文
C2 - 17628732
AN - SCOPUS:34547607954
SN - 0168-0102
VL - 58
SP - 349
EP - 355
JO - Neuroscience Research
JF - Neuroscience Research
IS - 4
ER -