Belayev L, Busto R, Zhao W, Fernandez G, Ginsberg M D
Cerebral Vascular Disease Research Center, Department of Neurology (D4-5), University of Miami School of Medicine, PO Box 016960, Miami, FL 33101, USA.
Brain Res. 1999 Jul 3;833(2):181-90. doi: 10.1016/s0006-8993(99)01528-0.
The present study was conducted to validate a modified method of temporary focal cerebral ischemia in the mouse; neurobehavioral function and histopathological infarction were quantitated following various periods of middle cerebral artery occlusion (MCAo). Male C57BL/6 mice were anesthetized with 3% halothane in a mixture of 30%O2/70%N2O delivered by face mask and were subjected to 30- to 180-min of temporary middle cerebral artery occlusion (MCAo) by an intraluminal suture coated with poly-l-lysine. Twenty-eight of 40 mice showed an initial high-grade neurological deficit (30-min MCAo, n=7; 60-min, n=8; 120-min, n=8; 180-min, n=5) when examined during MCAo; these were used for subsequent study. One day after MCAo, behavioral function was re-evaluated, and brains were perfusion-fixed and infarct volumes were measured. The initial neurological deficit improved at 24 h in mice with 30- or 60-min of prior MCAo but tended to persist in mice with 120- or 180-min insults. Following each duration of ischemia, mice exhibited ipsilateral infarcts. Small, inconsistent predominantly subcortical infarcts were present after 30-min MCAo, while longer occlusion periods gave rise to consistent foci of subcortical infarction involving striatum, septum, thalamus, and hippocampus, as well as areas of frontoparietal cortical infarction. The major advantages of the improved intraluminal MCAo model reported here, incorporating sutures coated with poly-l-lysine, include: a 100% incidence of infarction of predictable location and size in mice having an initial neurological deficit. Periods of 60- to 180-min MCA occlusion in this model yield sufficiently reproducible sequelae to permit the effects of various therapeutic agents on neurological outcome and size of infarction to be readily studied.
本研究旨在验证一种改良的小鼠短暂性局灶性脑缺血方法；在大脑中动脉闭塞（MCAo）不同时间段后，对神经行为功能和组织病理学梗死情况进行定量分析。雄性C57BL/6小鼠通过面罩吸入含3%氟烷的30%O2/70%N2O混合气体进行麻醉，然后通过涂有聚-L-赖氨酸的腔内缝合线进行30至180分钟的短暂大脑中动脉闭塞（MCAo）。在MCAo期间检查时，40只小鼠中有28只出现初始重度神经功能缺损（30分钟MCAo，n = 7；60分钟，n = 8；120分钟，n = 8；180分钟，n = 5）；这些小鼠用于后续研究。MCAo后一天，重新评估行为功能，对大脑进行灌注固定并测量梗死体积。先前MCAo 30或60分钟的小鼠在24小时时初始神经功能缺损有所改善，但先前MCAo 120或180分钟的小鼠中神经功能缺损往往持续存在。在每个缺血时间段后，小鼠均出现同侧梗死。30分钟MCAo后出现小的、不一致的主要为皮质下梗死，而较长的闭塞时间则导致涉及纹状体、隔区、丘脑和海马以及额顶叶皮质梗死区域的一致的皮质下梗死灶。此处报道的改良腔内MCAo模型的主要优点包括：初始有神经功能缺损的小鼠梗死发生率为100%，梗死位置和大小可预测。该模型中60至180分钟的MCA闭塞时间段产生的后遗症具有足够的可重复性，便于轻松研究各种治疗药物对神经功能结局和梗死大小的影响。
