Early nuclear translocation of endonuclease G and subsequent DNA fragmentation after transient focal cerebral ischemia in mice
Early nuclear translocation of endonuclease G and subsequent DNA fragmentation after transient focal cerebral ischemia in mice
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DOI：
10.1016/j.neulet.2005.05.058
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发表时间：
2005-09-01
影响因子：
2.5
通讯作者：
Kim, GW
中科院分区：
文献类型：
--
作者：
Lee, BI;Lee, DJ;Kim, GW
我们研究了短暂性局灶性脑缺血(TFCI)后，内切酶G(Endog)是否从线粒体移位到细胞核，从而导致随后的DNA片段化。成年雄性小鼠采用线栓法阻断大脑中动脉造成60min局灶性脑缺血模型。在tFCL的不同时间点进行endog蛋白免疫印迹分析，在tFCI后4h即可检测到线粒体endog的表达，再灌注4h线粒体endog明显减少(p<；0.01)。Endog免疫组织化学证实，早在tFCI后4h，即可在缺血区大脑中动脉(MCA)区域检测到EnDog的核移位。EnDog和AIF双重免疫荧光染色显示，在tFCI后24 h，Endog主要与AIF共存。Endog免疫组织化学和TdT介导的dUTP-生物素缺口末端标记双重染色显示，在tFCI后24 h，Endog表达与DNA片段化之间存在空间关系。这些数据表明，在tFCI后，脑缺血后早期发生了endog的核移位，并可能导致DNA片段化。(C)2005爱思唯尔爱尔兰有限公司。保留所有权利。
We investigated whether the endonuclease G (endoG) translocated from mitochondria to nucleus after transient focal cerebral ischemia (tFCI), thereby contributed to subsequent DNA fragmentation. Adult male mice were subjected to 60 min of focal cerebral ischemia by intraluminal suture blockade of the middle cerebral artery. Western blot analysis for endoG was performed at various time points of tFCL Nuclear endoG was detected as early as 4 h after tFCI in the ischemic brain, and correspondingly mitochondrial endoG showed a significant reduction at 4 h after reperfusion (p < 0.01). Immunohistochemistry of endoG confirmed that the nuclear translocation of endoG was detected as early as 4 h after tFCI in the middle cerebral artery (MCA) territory of the ischemic brain. Double immunofluorescent staining with endoG and AIF showed that endoG was predominantly colocalized with AIF at 24 h after tFCI. Double staining with endoG immunohistochemistry and TdT-mediated dUTP-biotin nick end labeling showed a spatial relationship between endoG expression and DNA fragmentation at 24 h after tFCI. These data suggest that the early nuclear translocation of endoG occurs and could induce DNA fragmentation in the ischemic brain after tFCI. (C) 2005 Elsevier Ireland Ltd. All rights reserved.