该协议被批准的机构动物护理和使用委员会（IACUC）在佛罗里达大学，并遵守的“实验动物保健的原则”（美国国立卫生研究院出版号86-23，1985年修订）。 1。材料<ol><li>动物：雄性C57BL / 6小鼠（哈伦实验室，印第安纳波利斯，在美国）在手术时重达20-26克。 </li><li>麻醉： <ol><li>麻醉机：VetEquip吸入麻醉系统（VetEquip公司，Pleasanton的，CA，美国）。 </li><li>吸入麻醉药异氟醚（百特制药，迪尔菲尔德，IL）。 </li></ol></li><li>手术工具：SuperCut虹膜剪刀，直（世界精密仪器公司，美国，佛罗里达州，萨拉索塔）; Vannas微型剪刀，直（世界精密仪器公司，美国，佛罗里达州，萨拉索塔）;杜蒙钳（精尖， 45 °弯曲的技巧，世界精密仪器公司，美国，佛罗里达州，萨拉索塔），双极电凝单元（高频干燥器900，世界精密仪器公司，萨拉索塔，佛罗里达州，美国）;微型血管夹（世界精密仪器，公司，美国，佛罗里达州，萨拉索塔） </li><li>手术用品：无菌棉提示;湿巾，纱布海绵，缝合（7-0丝线缝合，爱惜康公司，萨默维尔，新泽西州，美国）;组织粘合剂（Vetbond，3M公司，圣保罗，明尼苏达，美国）;手术擦洗Bethadine，70％的乙醇和无菌0.9％生理盐水;眼膏（Puralube兽医软膏，Pharmaderm，梅尔维尔，纽约，美国）。 </li><li>闭塞缝合准备缺血： <ol><li> MCA闭塞长丝：7-0手术尼龙单丝缝合（爱惜康公司，萨默维尔，新泽西州，美国）。 </li><li>钝化缝合头（热烧灼单位，世界精密仪器公司，萨拉索塔，佛罗里达州）的热源。 </li><li>液体硅橡胶（美国佛罗里达州萨拉索塔，有机硅室温胶，世界精密仪器公司，）。 </li></ol></li><li>外科/解剖显微镜（徕卡立体显微镜MZ6）。 <ol><li>光纤照明（光纤照明FO1 - 150，TechniQuip公司，利弗莫尔，CA，美国）。 </li></ol></li><li>激光多普勒血流计（5000，斯德哥尔摩，瑞典，Perimed PeriFlux系统）。 <ol><li>灵活的微尖（MT B500 - 0直微尖，Perimed，斯德哥尔摩，瑞典） </li></ol></li><li>温度参数： <ol><li>直肠温度监测（BAT - 12，世界精密仪器公司，萨拉索塔，佛罗里达州）。 </li><li>隔热毯（T /泵，TP600，Gaymar，纽约，美国）。 </li></ol></li><li>染色和定量测量体积梗死： <ol><li> 2％的溶液2,3,5 - 三苯基氯化（2％TTC，Sigma - Aldrich公司有限公司，圣路易斯，密苏里州，美国）。 </li><li>小鼠脑矩阵切片（Zivic米勒实验室，公司艾里逊公园，PA，美国）。 </li><li>数码相机（V1253，柯达公司，罗彻斯特，纽约州，美国）。 </li><li> Image J软件（ImageJ 1.42q软件，美国国立卫生研究院，贝塞斯达，MA，USA）。 </li></ol></li></ol> 2。手术前的步骤<ol><li>手术前，小鼠被安置在一个12:12的光/暗周期免费获得水和灭鼠议员。 </li><li>缺血缝线钝化为1 - 2mm的热和其与液态硅橡胶的尖端3 - 4mm的缝合轴涂层缝合提示准备，允许它干（7-0;尖端直径0.20 -0.25毫米;涂层长3 - 4mm的缝合长度为11 - 13MM）。不孕症应予以保留，在上述所有步骤。 </li><li>称重鼠标后，麻醉诱导用4％的异氟醚在感应室内保持在1.5-2％，30％O 2和70％N 2 O的混合气体通过口罩异氟醚的程序。 </li><li>鼠标被放置在俯卧姿势趴在一个温度控制的作业面（隔热毯）上的纱布垫;剃光头皮毛。 </li><li>应用润滑剂眼药膏双眼，眼睑闭合，以防止在手术过程中的眼干燥。 </li><li>一个正中切口（1厘米）的皮肤上覆的颅骨上项线到鼻根皮肤拉横向贴上一个灵活的微尖（MT B500 - 0直微尖，直径0.5mm）。微尖是垂直放置右顶叶头骨表面（后1毫米和5毫米，前囟门外侧），监测激光多普勒血流计血流在MCA领土。 </li><li>确保灵活的微尖是颅骨，坚定地前动物转向仰卧位。动物的脖子上放置一个软垫，它的鼻子是在鼻锥麻醉设备举行。这将导致有足够的空间，灵活的微尖。 </li><li>直肠温度探头插入，以保持恒定的动物核心温度为37 ± 0.5 ° C。 </li><li>这两个前爪和hindpaws录音操作平台表面，和尾巴是贴在直肠管。 </li> &lt;/ OL&gt; 3。 MCA闭塞手术： <ol><li>中线颈部切口手术显微镜下，用剪刀柄和下颚之间，轻轻提起皮肤松弛。 </li><li>配对的基础颌下腺直言不讳地划分，留在原地左腺。右颌下腺缩回颅，沿胸锁乳突肌和担保。 </li><li>肩胛舌骨肌覆盖颈动脉鞘暴露分工仔细划分与皮卡。 </li><li>仔细分离右侧颈总动脉（CCA），并从中分离出的迷走神经和鞘（位于外侧共同国家评估）。 </li><li> 此后，2，经常被用来执行MCA闭塞的手术方法不同 ： </li></ol>小泉的4方法<ol class="lroman" type="i"><li>右CCA是围绕着三个宽松的领口缝合（7-0丝绸）。 </li><li>颈外动脉（ECA）是确定的，孤立的，和烧灼（2 - 3mm的远端分叉）。随后，它可以分为微型剪刀。为了避免不慎周边损害烧灼网站，电源应设置在1使用电源旋钮。这是所有cauterizations一致。 </li><li>足够的张力是适用于对CCA最领缝合近端闭塞的血流量（永久性结扎）。流闭塞验证了〜基线皮质灌注值下降50％。 </li><li>然后一个血管夹是放在前文建会分岔动脉切开术的时间，以避免在逆行流。 </li><li>动脉切开术之间的近端结扎缝合，并用微型剪刀（靠近结扎）的船只上的CCA剪辑。 </li><li>热迟钝硅涂层7-0尼龙缝线是通过动脉切开术和引入先进的，直到它到达血管夹。 2其余宽松的领口缝合轻轻收紧各地的CCA之间通过共同国家评估，以避免流漏，谨慎，不创伤动脉壁的动脉切开术和血管夹，然后血管夹被撤回。 </li></ol>龙格的方法3 <ol class="lalpha" type="a"><li>非洲经济委员会鉴定，解剖尽可能（近到语言和上颌动脉的分叉处），烧灼（设置为1的电源），分为微型剪刀。 </li><li>是围绕着以上的文建会分叉近端ICA的两个宽松的领口缝合（7-0丝绸）。 </li><li>文建会暂时封闭，用血管夹。流闭塞验证了〜基线皮质灌注值下降50％。 </li><li> ICA是暂时关闭与上述两个宽松的领口缝合血管夹，动脉切开术的时间，以避免在逆行流。 </li><li>动脉切开术是使用微型剪刀，在反映非洲经委会密切残端。 </li><li>热迟钝硅涂层7-0尼龙缝线是通过动脉切开术在非洲经委会和引入先进的，直到它到达应用血管夹在ICA。近端ICA的2宽松的领口缝合近端ICA周围轻轻地拧紧，以避免通过ICA回流血流足够的重视，没有创伤的动脉壁，血管夹，然后被撤回。 </li></ol>该过程的其余部分主要是收敛： <ol><li>正在慢慢走向颅底颈内动脉（ICA）的通过先进的咬合缝合，直到感到一种轻微的抵抗。一个并发下降一个百分点（= 80％），相对于基线脑血流量值是用激光多普勒血流仪证实。 2领口缝合收紧周围插入长丝。 </li><li>短暂性脑缺血的实验，45分钟后，轻轻咬合缝合撤回。这个时间点，可以改变各种实验的目的。 </li><li>撤回缝合长丝后，动脉切开术的网站被关闭，以避免手术后出血。在小泉的方法，这涉及到各地的CCA安全收紧的领口缝合。龙格的技术，在非洲经委会的基础动脉切开术网站是烧灼，然后松开领口缝合近端ICA和删除，以恢复颈动脉流。 </li><li>要关闭切口，下颌下腺和SCM奠定了手术领域。伤口边缘，然后在宫颈癌组织粘合剂和头皮地区近似。另外，可吸收缝合线或金属主食可以用来关闭，特别是当动物的长期生存的目的是伤口。 </li><li>关闭切口后，动物收到温生理盐水1毫升皮下注射量补货。应使用一个适当的止痛药的剂量（即丁丙诺啡在0.05-0.1毫克/公斤），以尽量减少在恢复期动物的疼痛和不适。阴性特质L是从手术套房，并放置在一个温暖的恢复面积/笼，以防止低温，自由，方便地访问柔软的食物和水。 </li></ol> 4。术后神经功能评价动物恢复后总的意识，神经功能缺损可以通过一个简单的规模（五点量表） 评估如下3： <ol start="0"><li>没有观察到的赤字</li><li>故障，延长对侧前爪（轻度局灶性神经功能缺损） </li><li>绕在一个方向对侧梗死（中度局灶性神经功能缺损） </li><li>在一个方向对侧坠落，以梗死（严重的局灶性神经赤字） </li><li>郁闷的意识水平，无自发运动</li></ol> 5。染色及脑梗死体积的定量测量<ol><li>马华再灌注24小时后，幸存的小鼠称重，深为4％isoflorane麻醉，并根据批准的协议牺牲。 </li><li>大脑是小心取出，放置在小鼠大脑矩阵切片，和整个大脑是在2毫米间隔冠状切片。 </li><li>部分都沉浸在2％TTC磷酸缓冲液（PBS）在37 ° 20分钟。 TTC染色后，梗死面积是白色（未染色）组织相邻的红砖（可行）组织（图1）所示。 </li><li>连续切片拍摄使用数码相机和梗死面积测量与图像J计划。 </li><li>梗塞面积（白，不染），同侧半球面积（白色和红色的砖，染色），和对侧半球的面积（红砖，彩色），测量每个节。体积的计算方法是在所有路段的总结有代表性的地区和乘以层厚。水肿纠正后，梗死体积的计算方法如下9： 更正梗死体积（CIV），％= [对侧大脑半球额- （同侧半球体积-梗塞体积）] /对侧大脑半球体积× 100 </li></ol> 6。代表性的成果后，左文建会闭塞，并通过建立短暂性脑缺血再灌注MCA的，动物的牺牲和他们的大脑冠状切片，用TTC染色评价梗死体积（图1）。 <img src="files/ftp_upload/2879/2879fig1.jpg" alt="图1" /> 图1。TTC染色串行冠状从小鼠的脑切片（2毫米）MCA闭塞。

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作者宣布，他们已经没有利益冲突。

MCAO模型是一个既定的模式，复制脑缺血小鼠物种。第一次使用这个模型是在大鼠和以后适应小鼠。避免去骨瓣，以消除对血脑屏障通透性和颅内压手术操作的影响，是该技术的优点。然而，许多变量，如应变相关的差异 10，大小缝合技巧 11，持续时间 6闭塞， 体温 12， 麻醉 13，以及其他因素，可能导致梗塞体积在这个模型不一致。 值得一提的有具?...

<table border="1"><tbody><tr><th>试剂名称</th><th>类型</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td>雄性C57BL / 6小鼠</td><td>动物</td><td>哈伦实验室，印第安纳波利斯，在美国</td><td></td><td> 20-26 GRS </td></tr><tr><td>解剖镜下</td><td>显微镜</td><td>徕卡立体显微镜MZ6 </td><td></td><td> 6.3:1变焦</td></tr><tr><td>光纤光纤照明</td><td>照明灯</td><td> TechniQuip公司，利弗莫尔，CA，美国</td><td></td><td> FO1 - 150 </td></tr><tr><td>异氟醚</td><td>麻醉</td><td>巴克斯特制药，迪尔菲尔德，白细胞介素</td><td> 1001936060 </td><td></td></tr><tr><td> SuperCut虹膜剪，直</td><td>手术工具</td><td>世界精密仪器公司，美国，佛罗里达州，萨拉索塔</td><td> 14218 </td><td></td></tr><tr><td> Vannas微，剪刀，直</td><td>手术工具</td><td>世界精密仪器公司，美国，佛罗里达州，萨拉索塔</td><td> 14003 </td><td></td></tr><tr><td>杜蒙细尖镊子</td><td>手术工具</td><td>世界精密仪器公司，美国，佛罗里达州，萨拉索塔</td><td> 503290 </td><td></td></tr><tr><td>血管夹</td><td>手术工具</td><td>世界精密仪器公司，美国，佛罗里达州，萨拉索塔</td><td> 14120 </td><td></td></tr><tr><td>高频干燥器900 </td><td>手术工具</td><td>世界精密仪器公司，美国，佛罗里达州，萨拉索塔</td><td> 500397 </td><td></td></tr><tr><td> 7-0手术尼龙单丝缝合</td><td>缝合</td><td>爱惜康公司，美国，新泽西州，的萨默维尔</td><td> 1647G </td><td>缝合闭塞</td></tr><tr><td> 7-0手术丝线缝合</td><td>缝合</td><td>爱惜康公司，美国，新泽西州，的萨默维尔</td><td> 7733G </td><td>缝合结扎</td></tr><tr><td>有机硅室温胶</td><td>硅橡胶</td><td>世界精密仪器公司，美国，佛罗里达州，萨拉索塔</td><td> 1571 </td><td></td></tr><tr><td> Vetbond </td><td>组织粘合剂</td><td> 3M公司，美国明尼苏达州圣保罗， </td><td> 70200746587 </td><td></td></tr><tr><td> Puralube兽医软膏</td><td>眼膏</td><td> Pharmaderm，梅尔维尔，纽约，美国</td><td></td><td></td></tr><tr><td>丁丙诺啡hydrochlorode </td><td>止痛药</td><td> Hospira公司公司，森林湖，IL，美国</td><td> NDC 0409-2012-32 </td><td></td></tr><tr><td>热烧灼组</td><td></td><td>世界精密仪器公司，美国，佛罗里达州，萨拉索塔</td><td> 501292 </td><td></td></tr><tr><td> PeriFlux系统5000 </td><td>经颅多普勒血流</td><td> Perimed，斯德哥尔摩，Sweeden </td><td></td><td></td></tr><tr><td> BAT - 12 </td><td>直肠温度计</td><td>世界精密仪器公司，美国，佛罗里达州，萨拉索塔</td><td>英美烟草- 12R </td><td></td></tr><tr><td> T /泵，TP600 </td><td>隔热毯</td><td>美国，纽约，Gaymar </td><td></td><td></td></tr><tr><td> 2,3,5 - 三苯基氯化</td><td>试剂</td><td> Sigma - Aldrich公司有限公司，美国密苏里州圣路易斯， </td><td> T8877 </td><td></td></tr><tr><td>小鼠脑矩阵切片机</td><td>脑切片</td><td> Zivic米勒实验室，公司艾里逊公园，PA，美国</td><td> BSMAS001 - 1 </td><td></td></tr></tbody></table>

intraluminal middle cerebral artery occlusion (mcao) model for ischemic stroke with laser doppler flowmetry guidance in mice

中风是第三大死因，在世界上的残疾的首要原因，美国在2009 年 1,2的近70亿美元，估计所需费用。腔内大脑中动脉闭塞（MCAO）模型，于1986年，以模拟在大鼠有影响力的人类病理学由小泉纯一郎 4 。缺血法的修改后由龙格 3 。这两种技术已被广泛用来识别脑损伤的分子机制，导致缺血性中风和潜在的治疗方式5。一直延续这相对无创的方法，在大鼠，小鼠的使用，利用转基因和基因敲除株6,7优势。局灶性脑缺血模型，线栓是先进的经颈内动脉闭塞MCA的基地。回缩后，在指定的时间内缝合模仿自发再灌注损伤，但缝合，也可永久保留。该视频将展示在小鼠进行腔内缺血过程在逐步时尚的两个主要方法，以及为潜在的弊端和缺陷的见解。缺血性脑组织也将随之由2,3,5 -三苯基氯化四氮唑（TTC）染色，评估脑梗塞 8的程度。

Watch this Scientific Journal Video about Intraluminal Middle Cerebral Artery Occlusion MCAO Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice at JoVE.com

Intraluminal Middle Cerebral Artery Occlusion MCAO Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

腔内大脑中动脉闭塞（MCAO）模型是最常见的缺血性中风的实验模型中使用的模型。在这里，我们将展示在整个模型用激光多普勒血流计指南的细节，其代表性的成果。

腔内中东大脑中动脉闭塞（MCAO）缺血性中风模型在小鼠体内的激光多普勒血流指导

Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion 
in the United ...

intraluminal-middle-cerebral-artery-occlusion-mcao-model-for-ischemic

Research

JoVE Journal

Medicine

Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

49.6K 次观看.  University of Florida. 腔内大脑中动脉闭塞（MCAO）模型是最常见的缺血性中风的实验模型中使用的模型。在这里，我们将展示在整个模型用激光多普勒血流计指南的细节，其代表性的成果。

视频: Intraluminal Middle Cerebral Artery Occlusion MCAO Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

Modeling Stroke in Mice - Middle Cerebral Artery Occlusion with the Filament Model

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are very limited. To meet the need for novel therapeutic approaches, experimental stroke research frequently employs rodent models of focal cerebral ischaemia. Most researchers use permanent or transient occlusion of the middle cerebral artery (MCA) in mice or rats.
Proximal occlusion of the middle cerebral artery (MCA) via the intraluminal suture technique (so called filament or suture model) is probably the most frequently used model in experimental stroke research. The intraluminal MCAO model offers the advantage of inducing reproducible transient or permanent ischaemia of the MCA territory in a relatively non-invasive manner. Intraluminal approaches interrupt the blood flow of the entire territory of this artery. Filament occlusion thus arrests flow proximal to the lenticulo-striate arteries, which supply the basal ganglia. Filament occlusion of the MCA results in reproducible lesions in the cortex and striatum and can be either permanent or transient. In contrast, models inducing distal (to the branching of the lenticulo-striate arteries) MCA occlusion typically spare the striatum and primarily involve the neocortex. In addition these models do require craniectomy. In the model demonstrated in this article, a silicon coated filament is introduced into the common carotid artery and advanced along the internal carotid artery into the Circle of Willis, where it blocks the origin of the middle cerebral artery. In patients, occlusions of the middle cerebral artery are among the most common causes of ischaemic stroke. Since varying ischemic intervals can be chosen freely in this model depending on the time point of reperfusion, ischaemic lesions with varying degrees of severity can be produced. Reperfusion by removal of the occluding filament at least partially models the restoration of blood flow after spontaneous or therapeutic (tPA) lysis of a thromboembolic clot in humans.
In this video we will present the basic technique as well as the major pitfalls and confounders which may limit the predictive value of this model.

Filamentous occlusion of the Middle cerebral artery is a common model for studying ischemic stroke in mice.

建模在小鼠体内的行程 - 中东与灯丝模型大脑中动脉闭塞

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are ...

科研

医学

Mouse Model of Middle Cerebral Artery Occlusion

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the brain (ischemic stroke) 2. Since most ischemic strokes (~80%) occur in the territory of middle cerebral artery (MCA) 3, many animal stroke models that have been developed have focused on this artery. The intraluminal monofilament model of middle cerebral artery occlusion (MCAO) involves the insertion of a surgical filament into the external carotid artery and threading it forward into the internal carotid artery (ICA) until the tip occludes the origin of the MCA, resulting in a cessation of blood flow and subsequent brain infarction in the MCA territory 4. The technique can be used to model permanent or transient occlusion 5. If the suture is removed after a certain interval (30 min, 1 h, or 2 h), reperfusion is achieved (transient MCAO); if the filament is left in place (24 h) the procedure is suitable as a model of permanent MCAO. This technique does not require craniectomy, a neurosurgical procedure to remove a portion of skull, which may affect intracranial pressure and temperature 6. It has become the most frequently used method to mimic permanent and transient focal cerebral ischemia in rats and mice 7,8. To evaluate the extent of cerebral infarction, we stain brain slices with 2,3,5-triphenyltetrazolium chloride (TTC) to identify ischemic brain tissue 9. In this video, we demonstrate the MCAO method and the determination of infarct size by TTC staining.

We demonstrate in the video a method for producing a middle cerebral artery occlusion in adult mice using an intraluminal monofilament. We also show how to evaluate the extent of cerebral infarction by 2,3,5-triphenyltetrazolium chloride (TTC) staining.

大脑中动脉闭塞的小鼠模型

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the ...

Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We have established bilateral common carotid artery occlusion as a preconditioning stimulus to induce early ischemic tolerance to transient focal cerebral ischemia in C57Bl6/J mice. In this video, we will demonstrate the methodology used for this study.

There is accumulating evidence, that ischemic preconditioning (PC) &#x2013; a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We established bilateral common carotid artery occlusion (BCCAO) as a preconditioning stimulus to induce early ischemic tolerance (IT) to transient focal cerebral ischemia (induced by middle cerebral artery occlusion, MCAO) in C57Bl6/J mice.

双侧颈总动脉闭塞足够的预处理刺激诱导局灶性脑缺血早期缺血耐受

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a ...

Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in 20091, 2. Several models of cerebral ischemia have been developed to mimic the human condition of stroke. It has been suggested that up to 80% of all strokes result from ischemic damage in the middle cerebral artery (MCA) area3. In the early 1990s, endothelin-1 (ET-1) 4 was used to induce ischemia by applying it directly adjacent to the surface of the MCA after craniotomy. Later, this model was modified 5 by using a stereotaxic injection of ET-1 adjacent to the MCA to produce focal cerebral ischemia. The main advantages of this model include the ability to perform the procedure quickly, the ability to control artery constriction by altering the dose of ET-1 delivered, no need to manipulate the extracranial vessels supplying blood to the brain as well as gradual reperfusion rates that more closely mimics the reperfusion in humans5-7. On the other hand, the ET-1 model has disadvantages that include the need for a craniotomy, as well as higher variability in stroke volume8. This variability can be reduced with the use of laser Doppler flowmetry (LDF) to verify cerebral ischemia during ET-1 infusion. Factors that affect stroke variability include precision of infusion and the batch of the ET-1 used6. Another important consideration is that although reperfusion is a common occurrence in human stroke, the duration of occlusion for ET-1 induced MCAO may not closely mimic that of human stroke where many patients have partial reperfusion over a period of hours to days following occlusion9, 10. This protocol will describe in detail the ET-1 induced MCAO model for ischemic stroke in rats. It will also draw attention to special considerations and potential drawbacks throughout the procedure.

Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats. In addition, important considerations, advantages, and shortcomings of this model are discussed.

内皮素-1诱导大脑中动脉闭塞模型缺血性中风激光多普勒血流仪在大鼠指导

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in ...

Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be performed in many locations overlying the rat cerebrum, but is most easily carried out by creating a craniectomy over the readily accessible frontal or parietal bones. Most frequently, we have used this technique in combination with the endothelin-1 middle cerebral artery occlusion model of ischemic stroke to quantify the changes in middle cerebral artery vessel diameter that occur with injection of endothelin-1 into the brain parenchyma adjacent to the proximal MCA4, 5. In order to visualize the proximal portion of the MCA during endothelin -1 induced MCAO, we use a technique to create a cranial window through the temporal bone on the lateral aspect of the rat skull (Figure 1). Cerebral arteries can be visualized either with the dura intact or with the dura incised and retracted. Most commonly, we leave the dura intact during visualization since endothelin-1 induced MCAO involves delivery of the vasoconstricting peptide into the brain parenchyma. This bypasses the need to incise the dura directly over the visualized vessels for drug delivery. This protocol will describe how to create a cranial window to visualize cerebral arteries in a step-wise fashion, as well as how to avoid many of the potential pitfalls pertaining to this method.

This article describes a method for visualizing rat cerebral arteries through a cranial window using temporal craniectomy in order to view proximal portions of the middle cerebral artery (Figure 1). This versatile method can be combined with various techniques of drug delivery to measure cerebral artery reactivity in vivo.

利用颅窗口，在可视化的大脑中动脉内皮素-1诱导大脑中动脉闭塞

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be ...

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

优化系统的脑灌注监测在中风模型大鼠的大脑中动脉闭塞腔内

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

Modeling Stroke in Mice: Permanent Coagulation of the Distal Middle Cerebral Artery

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical options are available for a small proportion of stroke patients in the acute phase. Current research is intensively searching for novel therapeutic strategies and is increasingly focusing on the sub-acute and chronic phase after stroke because more patients might be eligible for therapeutic interventions in a prolonged time window. These delayed mechanisms include important pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity and regeneration. In order to analyze these mechanisms and to subsequently evaluate novel drug targets, experimental stroke models with clinical relevance, low mortality and high reproducibility are sought after. Moreover, mice are the smallest mammals in which a focal stroke lesion can be induced and for which a broad spectrum of transgenic models are available. Therefore, we describe here the mouse model of transcranial, permanent coagulation of the middle cerebral artery via electrocoagulation distal of the lenticulostriatal arteries, the so-called &#8220;coagulation model&#8221;. The resulting infarct in this model is located mainly in the cortex; the relative infarct volume in relation to brain size corresponds to the majority of human strokes. Moreover, the model fulfills the above-mentioned criteria of reproducibility and low mortality. In this video we demonstrate the surgical methods of stroke induction in the &#8220;coagulation model&#8221; and report histological and functional analysis tools.

Various murine models of middle cerebral artery occlusion (MCAO) are widely used in experimental brain research. Here, we demonstrate the model of transcranial permanent distal MCAO which produces consistent cortical infarction of a size corresponding to damage imposed by the majority of human ischemic strokes.

造型中风小鼠：在远端大脑中动脉永久性混凝

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical ...

Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic stroke. However, the use of tPA is limited by its narrow therapeutic window and by increased risk of hemorrhagic transformation. There is an urgent need to develop suitable stroke models to study new thrombolytic agents and strategies for treatment of ischemic stroke. At present, two major types of ischemic stroke models have been developed in rats and mice: intraluminal suture MCAO and embolic MCAO. Although MCAO models via the intraluminal suture technique have been widely used in mechanism-driven stroke research, these suture models do not mimic the clinical situation and are not suitable for thrombolytic studies. Among these models, the embolic MCAO model closely mimics human ischemic stroke and is suitable for preclinical investigation of thrombolytic therapy. This embolic model was first developed in rats by Overgaard et al.1 in 1992 and further characterized by Zhang et al. in 19972. Although embolic MCAO has gained increasing attention, there are technical problems faced by many laboratories. To meet increasing needs for thrombolytic research, we present a highly reproducible model of embolic MCAO in the rat, which can develop a predictable infarct volume within the MCA territory. In brief, a modified PE-50 tube is gently advanced from the external carotid artery (ECA) into the lumen of the internal carotid artery (ICA) until the tip of the catheter reaches the origin of the MCA. Through the catheter, a single homologous blood clot is placed at the origin of the MCA. To identify the success of MCA occlusion, regional cerebral blood flow was monitored, neurological deficits and infarct volumes were measured. The techniques presented in this paper should help investigators to overcome technical problems for establishing this model for stroke research.

Embolic Middle Cerebral Artery Occlusion MCAO for Ischemic Stroke with Homologous Blood Clots in Rats

In this paper, we demonstrate a standard method for producing an embolic middle cerebral artery occlusion with homologous blood clots (fibrin-rich) in adult rat. This model closely mimics human ischemic stroke and is suitable for preclinical study of thrombolytic therapy for ischemic stroke.

栓塞性脑梗塞（缺血）的大鼠缺血性中风与血同源凝块

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic ...

Surgical Approach for Middle Cerebral Artery Occlusion and Reperfusion Induced Stroke in Mice

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ischemic in origin and occur in the territory of the middle cerebral artery (MCA). Currently the only Food and Drug Administration (FDA) approved drug for the treatment of this devastating disease is tissue plasminogen activator (tPA). However, tPA has a small therapeutic window for administration (3 - 6 hr), and is only effective in 4% of the patients who actually receive it. Current research focuses on understanding the pathophysiology of stroke in order to find potential therapeutic targets. Thus, reliable models are crucial, and the MCA occlusion (MCAo) model (also termed the intraluminal filament or suture model) is deemed to be the most clinically relevant surgical model of ischemic stroke, and is fairly non-invasive and easily reproducible. Typically the MCAo model is used with rodents, especially with mice due to all the genetic variations available for this species. Here we describe (and present in the video) how to successfully perform the MCAo model (with reperfusion) in mice to generate reliable and reproducible data.

In order to understand the pathophysiology of stroke, it is important to use reliable models. This paper will describe one of the most frequently used stroke models in mice, termed the middle cerebral artery occlusion (MCAo) model (also termed the intraluminal filament or suture model) with reperfusion.

对大脑中动脉闭塞再灌注损伤诱发中风小鼠的手术方法

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ...

Induction of Ischemic Stroke and Ischemia-reperfusion in Mice Using the Middle Artery Occlusion Technique and Visualization of Infarct Area

Cerebrovascular disease is highly prevalent in the global population and encompasses several types of conditions, including stroke. To study the impact of stroke on tissue injury and to evaluate the effectiveness of therapeutic interventions, several experimental models in a variety of species were developed. They include complete global cerebral ischemia, incomplete global ischemia, focal cerebral ischemia, and multifocal cerebral ischemia. The model described in this protocol is based on the middle cerebral artery occlusion (MCAO) and is related to the focal ischemia category. This technique produces consistent focal ischemia in a strictly defined region of the hemisphere and is less invasive than other methods. The procedure described is performed on mice, given the availability of several genetic variants and the high number of tests standardized for mice to aid in the behavioral and neurodeficit evaluation.

We describe a mouse model of stroke induced by the occlusion of the middle cerebral artery using a silicone coated suture. The protocol can be applied to induce permanent occlusion or a temporary ischemia, followed by reperfusion.

缺血性中风和缺血再灌注小鼠的诱导使用梗塞区的中心动脉闭塞技术和可视化

Cerebrovascular disease is highly prevalent in the global population and encompasses several types of conditions, including stroke. To study the impact of ...