Sugimori Hiroshi, Yao Hiroshi, Ooboshi Hiroaki, Ibayashi Setsuro, Iida Mitsuo
Department of Medicine, Seiai Rehabilitation Hospital and Fukuoka Institute of Neurogenetics and Stroke, Onojo City, Fukuoka, Japan.
Brain Res Brain Res Protoc. 2004 Aug;13(3):189-96. doi: 10.1016/j.brainresprot.2004.06.001.
Recent advances in genetical engineering of the mouse have highlighted the importance of reproducible and less invasive models of cerebral ischemia in mice. In this paper, we developed minimally invasive and reproducible model of distal middle cerebral artery (MCA) occlusion in mice using krypton (Kr) laser-induced photothrombosis. C57BL/6 or BALB mice (n=8 each) were anesthetized with halothane. The skin was cut, the temporal muscle was retracted, and the right distal MCA was observed through the skull. A Kr laser beam of wavelength 568 nm was focused onto the MCA over the intact skull. Upon laser irradiation, intravenous administration of a rose bengal solution was begun. After 4 min of irradiation, the laser beam was refocused on the MCA just proximal to the first spot, and another 4-min irradiation was performed. Then, the right common carotid artery (CCA) was ligated. Three days later, the brain was removed, and infarct volume was determined. Infarction confined almost solely to the cortical area was produced in each mouse. Mean infarct volume in C57BL/6 mice was 25.2+/-13.7 mm3. The BALB mice group showed significantly larger and more reproducible infarction (44.1+/-5.2 mm3; the coefficient of variation was 12%) than did C57BL/6 mice (P<0.005). Our photothrombosis model of stroke in mice can be performed without craniectomy, and its reproducibility is satisfactory when using BALB mice.
小鼠基因工程的最新进展凸显了可重复且侵入性较小的小鼠脑缺血模型的重要性。在本文中,我们使用氪(Kr)激光诱导的光血栓形成法,开发了一种微创且可重复的小鼠大脑中动脉(MCA)远端闭塞模型。将C57BL/6或BALB小鼠(每组n = 8只)用氟烷麻醉。切开皮肤,牵开颞肌,通过颅骨观察右侧MCA远端。将波长为568 nm的Kr激光束聚焦在完整颅骨上方的MCA上。激光照射时,开始静脉注射孟加拉玫瑰红溶液。照射4分钟后,将激光束重新聚焦在第一个光斑近端的MCA上,再进行4分钟照射。然后,结扎右侧颈总动脉(CCA)。三天后,取出大脑并测定梗死体积。每只小鼠均产生几乎仅局限于皮质区域的梗死。C57BL/6小鼠的平均梗死体积为25.2±13.7 mm³。与C57BL/6小鼠相比,BALB小鼠组显示出明显更大且更具可重复性的梗死(44.1±5.2 mm³;变异系数为12%)(P<0.005)。我们的小鼠光血栓形成性中风模型无需开颅即可进行,并且在使用BALB小鼠时其可重复性令人满意。