DNA fragmentation in ischemic core and penumbra in focal cerebral ischemia in rats

被引:57
作者
Yao, H
Takasawa, R
Fukuda, K
Shiokawa, D
Sadanaga-Akiyoshi, F
Ibayashi, S
Tanuma, S
Uchimura, H
机构
[1] Kyushu Univ, Grad Sch Med Sci, Dept Med & Clin Sci, Dept Internal Med 2,Higashi Ku, Fukuoka 8128582, Japan
[2] Tokyo Univ Sci, Fac Pharmaceut Sci, Dept Biochem, Tokyo 162, Japan
[3] Hizen Natl Hosp, Neurochem Lab, Saga, Japan
来源
MOLECULAR BRAIN RESEARCH | 2001年 / 91卷 / 1-2期
关键词
apoptosis; cerebral ischemia; stroke; cerebral blood flow;
D O I
10.1016/S0169-328X(01)00135-8
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
Although apoptotic cell death has been suggested to be involved in ischemic injury of the brain, the precise mechanisms of ischemic neuronal cell death are unknown. Here, we examined the biochemical feature of apoptosis (i.e. DNA fragmentation) in male spontaneously hypertensive rats (5-7 months old) subjected to photothrombotic distal middle cerebral artery (MCA) occlusion. After MCA occlusion, the brain was cut in a cryostat to produce a standard coronal block and samples were dissected from the regions corresponding to the ischemic core, penumbra and contralateral control areas. Changes in cerebral blood Row (CBF) were monitored at I mm posterior and 2-4 mm lateral to the bregma by means of a laser-Doppler flowmetry. After MCA occlusion, CBF was decreased to 72 +/- 18 (+/-S.D.), 50 +/- 14, and 35 +/- 11% of the control values at 2, 3, and 4 mm from the midline, respectively. DNA fragmentation characteristics of apoptosis were examined in these samples by conventional and pulse-field gel electrophoresis. On the conventional gel electrophoresis, nucleosomal DNA fragmentation was detected in the penumbral zone at 6 h after MCA occlusion. Large DNA fragments of 50 and 20 kbp were detected in the penumbral zone and also in the ischemic core region at 3 h after distal MCA occlusion. The large DNA fragments seen on the pulse-field gel elecrophoresis were further degraded to small DNA fragments at 6 h after MCA occlusion in the penumbral zone but not in the core regions. The evolving DNA fragmentation was observed between 3 and 6 h after the onset of brain ischemia in the penumbra, suggesting that apoptosis may contribute to the development of ischemic infarction. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:112 / 118
页数:7
相关论文
共 28 条
[1]   CLEAVAGE OF DNA TO LARGE KILOBASE PAIR FRAGMENTS OCCURS IN SOME FORMS OF NECROSIS AS WELL AS APOPTOSIS [J].
BICKNELL, GR ;
COHEN, GM .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1995, 207 (01) :40-47
[2]   Photothrombotic middle cerebral artery occlusion in spontaneously hypertensive rats - Influence of substrain, gender, and distal middle cerebral artery patterns on infarct size [J].
Cai, H ;
Yao, H ;
Ibayashi, S ;
Uchimura, H ;
Fujishima, M .
STROKE, 1998, 29 (09) :1982-1986
[3]   Ultrastructural morphological changes are not characteristic of apoptotic cell death following focal cerebral ischaemia in the rat [J].
Campagne, MV ;
Gill, R .
NEUROSCIENCE LETTERS, 1996, 213 (02) :111-114
[4]   EARLY ENDONUCLEASE ACTIVATION FOLLOWING REVERSIBLE FOCAL ISCHEMIA IN THE RAT-BRAIN [J].
CHARRIAUTMARLANGUE, C ;
MARGAILL, I ;
PLOTKINE, M ;
BENARI, Y .
JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM, 1995, 15 (03) :385-388
[5]  
GINSBERG MD, 1995, CEREBROVASCULAR DISEASES, P331
[6]   IN-SITU DETECTION OF FRAGMENTED DNA (TUNEL ASSAY) FAILS TO DISCRIMINATE AMONG APOPTOSIS, NECROSIS, AND AUTOLYTIC CELL-DEATH - A CAUTIONARY NOTE [J].
GRASLKRAUPP, B ;
RUTTKAYNEDECKY, B ;
KOUDELKA, H ;
BUKOWSKA, K ;
BURSCH, W ;
SCHULTEHERMANN, R .
HEPATOLOGY, 1995, 21 (05) :1465-1468
[7]   Mitochondria and apoptosis [J].
Green, DR ;
Reed, JC .
SCIENCE, 1998, 281 (5381) :1309-1312
[8]   CEREBRAL BLOOD-FLOW AUTOREGULATION DURING HYPOBARIC HYPOTENSION ASSESSED BY LASER-DOPPLER SCANNING [J].
HEIMANN, A ;
KROPPENSTEDT, S ;
ULRICH, P ;
KEMPSKI, OS .
JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM, 1994, 14 (06) :1100-1105
[9]   AMPA receptor antagonist, YM90K, reduces infarct volume in thrombotic distal middle cerebral artery occlusion in spontaneously hypertensive rats [J].
Hiroshi, Y ;
Ibayashi, S ;
Nakane, H ;
Cai, H ;
Uchimura, H ;
Fujishima, M .
BRAIN RESEARCH, 1997, 753 (01) :80-85
[10]   VIABILITY THRESHOLDS AND THE PENUMBRA OF FOCAL ISCHEMIA [J].
HOSSMANN, KA .
ANNALS OF NEUROLOGY, 1994, 36 (04) :557-565