这项工作是支持由美国心脏协会大中华区东南亚联盟（09GRNT2060421），美国医学协会，并从大学的佛罗里达州的临床与转化科学研究所的资助。亚当麦加是一个NIH / NINDS，NRSA博士前研究员（F30 NS-060335）。高血压（T32 HL-083810）从佛罗里达州的多学科培训计划的大学，博士前奖学金罗伯特·Regenhardt收到支持。

机构动物管理和使用委员会（IACUC）在佛罗里达大学和批准该协议是符合"指南的照顾和使用的实验动物"（第八版，美国国家科学院，2011年）。 物料<ol><li>动物:8周龄，雄性，只SD大鼠（查尔斯河农场，威​​尔明顿，MA，USA），体重250〜300克，手术时间。 </li><li>麻醉<ol class="lalpha"><li>吸入麻醉系统（VetEquip公司，普莱森顿，CA，USA） </li><li>异氟醚麻醉剂（巴克斯特制药，迪尔菲尔德，IL，USA） </li></ol></li><li>立体定位系统（大卫KOPF工具，图洪加，CA，USA） <ol class="lalpha"><li>小动物立体定位系统</li><li>非破裂大鼠耳棒</li><li>气体麻醉大鼠头支架</li></ol></li><li>温度调节<ol class="lalpha"><li> BAT-12探针温度计（世界精密仪器公司，萨拉索塔，佛罗里达州，美国） </li><li> T / PUMP，TP600热毯（Gaymar工业公司，果树园公园，NY，USA） </li></ol></li><li>手术器械<ol class="lalpha"><li>梅岑鲍姆剪刀，虹膜钳，斗牛犬钳拉钩，10μl注射器，26号斜口针，Bovie烧灼试剂盒（世界精密仪器公司，萨拉索塔，佛罗里达州，美国） </li><li>微电机钻（Stoelting，伍德戴尔，IL，USA） </li><li> 0.8毫米的圆形钻牙钻（Roboz手术仪器有限公司，公司，盖瑟斯堡，MD，USA） </li><li> STORZ，波恩缝合钳（博士伦公司，罗切斯特，NY，USA） </li></ol></li><li>手术用品<ol class="lalpha"><li> 3.0尼龙线（绿洲，Mettawa，IL，USA） </li><li>棉拭子，Puralube的眼膏（默飞世尔科技，匹兹堡，PA，USA） </li><li>手术点和条（Xomed公司，杰克逊维尔，佛罗里达州，美国美敦力公司） </li><li>电推剪（奥斯特，普罗维登斯，RI，USA） </li></ol></lI&gt; <li>化学制品<ol class="lalpha"><li>内皮素-1（美国多肽，桑尼维尔，CA，USA） </li><li>洗必泰2％（Agrilabs，圣若瑟，MO，USA） </li></ol></li><li>可视化设备<ol class="lalpha"><li>手术显微镜（塞勒仪器及制造，圣路易斯，MO，USA） </li><li>索尼HDR-SR12（索尼，千代田区，东京，日本） </li><li>光纤照明器（TechniQuip公司，利弗莫尔，CA，USA） </li></ol></li><li>测量血管直径<ol class="lalpha"><li> VLC媒体播放器（巴黎，法国） </li><li> Image J软件（ImageJ的1.42q软件，美国国立卫生研究院，贝塞斯达，MA，USA） </li></ol></li></ol> 1。术前步骤<ol><li>手术前，老鼠被安置在一个12:12的光/暗周期，可免费使用水和啮齿动物州城。 </li><li>麻醉诱导用4％的异氟醚在100％O 2的混合气体的感应室中。 </li><li>乌鸦<html> n的头被剃电推剪。 </li><li>大鼠置于俯卧趴在一个温度控制的操作表面（隔热层）和头部被放置在与放置气体麻醉面罩的立体定向装置开始位置上的吸收垫。 </li><li>接着，耳棒被插入并拧紧。 </li><li>在手术过程中麻醉维持在100％O 2的混合气体，用2％异氟烷。 </li><li>眼科润滑软膏的双眼，眼皮被关闭，以防止眼睛干燥，在手术过程中。 </li><li>探针插入直肠温度为37±0.5℃下，以保持恒定的动物核心温度</li><li>与头牢固地保持在立体定位设备的麻醉大鼠的手术区是洁净其交替的洗必泰和盐水洗涤三次。 </li></ol> 2。前颅窗准备ontent颅创建一个窗口之前，应准备大鼠的任何其他所需的硬件植入实验，并应接受必要的外科手术。对于这个协议，我们先前已经植入了导套管内皮素-1（ET- 1）诱导脑缺血的配套出版物，题为"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。" 3。创建颅窗放置的导向套管或设备所需的实验后，颅窗口创建直接可视化在行程过程中的中脑动脉的近端部分。 <ol><li>首先，剪刀是用来覆盖开始向内侧颞肌和工作横向切开皮肤。 </li><li>颞肌使用电一分为二，然后缩回，用3.0尼龙线visua<html>利泽的颞骨鳞状部分。 </li><li>约3-4平方毫米的鳞状部分颞骨尾鳍的轨道和卓越的颧突的基础上绘制的，因为它反映的颞骨。 </li><li>使用的钻头，逐步削减的轮廓从颞骨的骨片。应小心避免钻头施加太大的压力，因为它有可能损坏硬脑膜或大脑皮质。 </li><li>经常用无菌生理盐水进行冲洗，提高外科领域的可视化和防止经济过热的头骨。 </li><li>在一个松散的角落，开始的一块的temoporal骨被小心取出用细鼠齿钳，同时确保不与硬脑膜撕裂任何船只。 </li><li>硬脑膜原封不动和碎片用无菌盐水冲洗掉。 </li></ol> 4。记录大脑中动脉收缩帐篷"&gt;为了演示如何捕获实时图像中，一个正在经历ET-1诱导脑缺血大鼠用于本协议。 <ol><li>血管活性化合物的应用程序之前，应该记录的基准视频至少1分钟。对于ET-1诱导缺血，基线记录一旦针已进入脑实质降低，但是之前的ET-1的注射。 </li><li>开始注射的注射器泵，并记录为1小时，或直至所需的端点。针在视频录制期间，留在原地为了防止干扰焦平面。 </li><li>根据批准的协议，此过程应该是深度麻醉大鼠和安乐死。 </li></ol> 5。图像分析可确定血管直径的可视化MCA的任何部分。作为一个例子，我们将使用的MCA的一个分支，测量血管的直径在时间点前后ET-1注射。性病会从视频帧被捕获在1分钟的时间间隔，使用VLC媒体播放器（VideoLAN的）。 <ol><li>安装VLC播放器并打开。 </li><li>通过选择工具，然后选择Preferences，将设置更改为"全部"下的"显示设置"。 </li><li> "视频"菜单"中展开左侧边栏和"输出模块"，然后扩大。 "场景过滤器"带来现场视频滤镜菜单中选择。 </li><li> "场景"中键入文件名前缀的框。 </li><li>目录路径前缀定义。这是保存静止帧。 </li><li>被选择的记录比基于静止帧捕获所需的速率。如果被抓获的视频每秒29帧，然后在"1749"（29帧/秒×60秒/分钟）应的方框中输入保存一个静止帧，每1分钟。所有的更改，然后保存。 </li><li>在VLC播放器，然后打开所需的视频自动保存静帧，每1分钟。 </li><li>然后，使用这些测量血管直径图像。首先，ImageJ软件（NIH）被打开。 </li><li>然后，你将测量的静帧，打开ImageJ的。 </li><li>选择"分析"菜单，"设置测量"，然后打开所有的箱子都没有选中。 </li><li>接着，将直线工具被选中。 </li><li>键盘快捷键的"Ctrl +"是用来在需要放大，一条线被置于垂直于容器路径测量血管直径。 </li><li>最后，船的长度是通过选择"分析"菜单，然后"措施"（Ctrl + M），以获得船只长度。 </li><li>这个过程被重复至少3次，取平均值为测量每个测量容器，在每个静止帧。 </li><li>被标准化为在每个时间点的血管直径，使基线血管直径的比较可以使用多个大鼠。要做到这一点，使用公式当前直径/基准直径×100％来计算的每艘船舶％的基准直径。 </li></ol>

<ol>
	<li>Levasseur, J. E., Wei, E. P., Raper, A. J., Kontos, A. A., Patterson, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detailed+description+of+a+cranial+window+technique+for+acute+and+chronic+experiments.">Detailed description of a cranial window technique for acute and chronic experiments.</a> Stroke. 6, 308-317 (1975).</li><li>Baumbach, G. L., Dobrin, P. B., Hart, M. N., Heistad, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanics+of+cerebral+arterioles+in+hypertensive+rats.">Mechanics of cerebral arterioles in hypertensive rats.</a> Circ. Res. 62, 56-64 (1988).</li><li>Regrigny, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+melatonin+on+rat+pial+arteriolar+diameter+in+vivo.">Effects of melatonin on rat pial arteriolar diameter in vivo.</a> Br. J. Pharmacol. 127, 1666-1670 (1999).</li><li>Mecca, A. P., O'Connor, T. E., Katovich, M. J., Sumners, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Candesartan+pretreatment+is+cerebroprotective+in+a+rat+model+of+endothelin-1-induced+middle+cerebral+artery+occlusion.">Candesartan pretreatment is cerebroprotective in a rat model of endothelin-1-induced middle cerebral artery occlusion.</a> Exp. Physiol. 94, 937-946 (2009).</li><li>Mecca, A. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebroprotection+by+angiotensin-(1-7)+in+endothelin-1-induced+ischaemic+stroke.">Cerebroprotection by angiotensin-(1-7) in endothelin-1-induced ischaemic stroke.</a> Exp. Physiol. 96, 1084-1096 (2011).</li></ol>

没有利益冲突的声明。

总之，这种颅窗制备技术是非常通用的，因为它可以被改变，以满足许多实验与细微的修改4，5。例如，我们已经成功地监测脑血流量在特定的MCA分支，使用激光多普勒血流仪集中在一个的颅骨窗口（麦加AP 2009和2011年）通过观察大脑中动脉。此外，与硬脑膜阴刻类似的制剂可以使用血管活性化合物的局部给药，以创建一个在体内血管反应槽3。有几个因素应予以考虑。制备?...

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td>吸入麻醉系统</td><td> VetEquip公司，美国加利福尼亚州Pleasanton， </td><td> 901806 </td><td></td></tr><tr><td>异氟醚麻醉</td><td>巴克斯特制药，迪尔菲尔德，IL，USA </td><td> 1001936060 </td><td></td></tr><tr><td>小动物立体定位系统</td><td>大卫KOPF仪器，图洪加，CA，USA </td><td> 900 </td><td></td></tr><tr><td>非破裂耳酒吧，大鼠</td><td>大卫KOPF仪器，图洪加，CA，USA </td><td> 957 </td><td></td></tr><tr><td>气体麻醉大鼠头支架</td><td>大卫KOPF仪器，图洪加，CA，USA </td><td> 1929年</td><td></td></tr><tr><td> BAT-12探针温度计</td><td>世界精密仪器公司，美国佛罗里达州，萨拉索塔， </td><td> BAT-12 </td><tD&gt; </td></tr><tr><td> T /泵，热毯</td><td> Gaymar工业公司，果树园公园，NY，USA </td><td> T / PUMP，TP600 </td><td></td></tr><tr><td>梅岑鲍姆剪刀</td><td>世界精密仪器公司，美国佛罗里达州，萨拉索塔， </td><td> 501254 </td><td></td></tr><tr><td>虹膜镊子</td><td>世界精密仪器公司，美国佛罗里达州，萨拉索塔， </td><td> 15915 </td><td></td></tr><tr><td>斗牛犬钳拉钩</td><td>世界精密仪器公司，美国佛罗里达州，萨拉索塔， </td><td> 14119-G </td><td></td></tr><tr><td> 10μl注射器26 gaugue </td><td>世界精密仪器公司，美国佛罗里达州，萨拉索塔， </td><td> SGE010RNS </td><td></td></tr><tr><td> Bovie，高温烧灼套件</td><td>世界精密仪器公司，美国佛罗里达州，萨拉索塔， </td><td> 500392 </td><td></td></tr><tr><td>鼠齿钳0.12 </td><td> Stotz </td><td> E1811 </td><td></td></tr><tr><td>微电机钻</td><td> Stoelting，伍德戴尔，IL，USA </td><td> 51449 </td><td></td></tr><tr><td> 0.8毫米的圆形钻牙钻</td><td> Roboz手术器械有限责任公司，美国马里兰州盖瑟斯堡， </td><td> RS-6280C-1 </td><td></td></tr><tr><td> STORZ波恩缝合钳</td><td>博士伦公司，罗切斯特，NY，USA </td><td></td><td></td></tr><tr><td>尼龙线，尺寸3.0 </td><td>绿洲，Mettawa，IL，USA </td><td> MV-663 </td><td></td></tr><tr><td>棉签</td><td>默飞世尔科技，匹兹堡，PA，USA </td><td> 22-029-488 </td><td></td></tr><tr><td> Puralube眼膏</td><td>默飞世尔科技，匹兹堡，PA，USA </td><td> NC0138063 </td><td></td></tr><tr><td>电推剪</td><td>奥斯特，普罗维登斯，RI，USA </td><td> 78005-301 </td><td></td></tr><tr><td> ET-1的80μM浓度在PBS中稀释至</td><td>美国多肽，桑尼维尔，CA，USA </td><td> 88-1-10A </td><td></td></tr><tr><td>洗必泰，2％; </td&gt; <td> Agrilabs，圣若瑟，密苏里州，美国</td><td> 1040，牧师6-06，NAC编号:10580322 </td><td></td></tr><tr><td>手术显微镜</td><td>塞勒仪器制造，圣路易斯，MO，USA </td><td>演化XR6 </td><td></td></tr><tr><td>索尼Handycam </td><td>索尼，日本东京都港区， </td><td> HDR-SR12 </td><td></td></tr><tr><td>光纤照明</td><td> TechniQuip公司，利弗莫尔，CA，USA </td><td> FO1-150 </td><td></td></tr><tr><td> VLC媒体播放器</td><td> （巴黎，法国） </td><td></td><td></td></tr><tr><td> Image J软件</td><td>美国国立健康，贝塞斯达，MA，USA </td><td></td><td></td></tr></tbody></table>

utilizing a cranial window to visualize the middle cerebral artery during endothelin-1 induced middle cerebral artery occlusion

颅窗口的创建是一个方法，可以直接可视化结构的大脑皮质表面1-3。这种技术可以覆盖的大鼠大脑在许多地方进行，但最容易地进行通过创建一个去骨瓣在容易获得额叶或顶叶骨头。最经常的是，我们已经使用了这种技术，组合与内皮素-1的大脑中动脉闭塞模型的缺血性中风的大脑中动脉的血管直径的变化所发生的脑实质与内皮素-1注入到相邻的近端MCA量化4，5。为了形象化在内皮素-1诱导的缺血的MCA的近端部分中，我们使用一种技术，以通过颞骨大鼠颅骨外侧面（ 图1）上建立一个颅窗。脑动脉可与硬脑膜完整或与硬脑膜切开retra的的可视化反恐执行局。最常见的是，我们离开硬脑膜完整过程的可视化，因为内皮素1诱导脑缺血涉及到脑实质内的血管收缩肽交付。这将绕过切开硬脑膜直接在可视化血管药物输送的需要。该协议将介绍如何创建一个颅窗的可视化脑动脉一个逐步的时尚，以及如何避免许多关于这种方法的潜在缺陷。

静止图像捕获的视频显示取自脑动脉直径的变化后ET-1注射可以容易地理解，使用此颅窗技术（ 图2）。 ET-1注射在几分钟之内，该船将开始收缩。最终，船只将是很难想象的脑组织会变得苍白。大约20分钟后，ET-1的影响将减少，血管开始扩张，逐渐恢复到基线直径约45分钟后。除了发生比较明显的血管收缩，ET-1后管理，皮质表面变得苍白。如果需要的话，它可以在容器直径计算的绝对...

Watch this Scientific Journal Video about Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion at JoVE.com

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

本文介绍了一种方法，可视化的大鼠大脑中动脉通过颅窗颞部颅骨切除术，以大脑中动脉近端部分（<strong&gt;图1</strong&gt;）。这种多用途的方法可以结合与药物输送的各种技术来测量脑动脉反应性<em在体内</em&gt;。

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 ...

utilizing-cranial-window-to-visualize-middle-cerebral-artery-during

Universidad de Cartagena UDEC

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JoVE Journal

Medicine

14.9K 次观看.  University of Florida. 本文介绍了一种方法，可视化的大鼠大脑中动脉通过颅窗颞部颅骨切除术，以大脑中动脉近端部分（图1）。这种多用途的方法可以结合与药物输送的各种技术来测量脑动脉反应性

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

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 ...

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

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 States in 20091,2. The intraluminal middle cerebral artery occlusion (MCAO) model was developed by Koizumi4 in 1986 to simulate this 
impactful human pathology in the rat. A modification of the MCAO method was later presented by Longa3. Both techniques have been widely used to identify 
molecular mechanisms of brain injury resulting from ischemic stroke and potential therapeutic modalities5. This relatively noninvasive method in rats has been 
extended to use in mice to take advantage of transgenic and knockout strains6,7. To model focal cerebral ischemia, an intraluminal suture is advanced via 
the internal carotid artery to occlude the base of the MCA. Retracting the suture after a specified period of time mimics spontaneous reperfusion, but the 
suture can also be permanently retained. This video will be demonstrating the two major approaches for performing intraluminal MCAO procedure in mice in a 
stepwise fashion, as well as providing insights for potential drawbacks and pitfalls. The ischemic brain tissue will subsequently be stained by 
2,3,5-triphenyltetrazolium chloride (TTC) to evaluate the extent of cerebral infarction8.

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

The intraluminal middle cerebral artery occlusion (MCAO) model is the most frequent used model among experimental ischemic stroke models. Here we will demonstrate the entire model in detail with the guide of Laser Doppler flowmetry, and its representative results.

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 ...

The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

Focal cerebral ischemia is among the most common type of stroke seen in patients. Due to the clinical significance there has been a prolonged effort to develop suitable animal models to study the events that unfold during ischemic insult. These techniques include transient or permanent, focal or global ischemia models using many different animal models, with the most common being rodents.
The permanent MCA ligation method which is also referred as pMCAo in the literature is used extensively as a focal ischemia model in rodents 1-6. This method was originally described for rats by Tamura et al. in 1981 7. In this protocol a craniotomy was used to access the MCA and the proximal regions were occluded by electrocoagulation. The infarcts involve mostly cortical and sometimes striatal regions depending on the location of the occlusion. This technique is now well established and used in many laboratories 8-13. Early use of this technique led to the definition and description of &#x201C;infarct core&#x201D; and &#x201C;penumbra&#x201D; 14-16, and it is often used to evaluate potential neuroprotective compounds 10, 12, 13, 17. Although the initial studies were performed in rats, permanent MCA ligation has been used successfully in mice with slight modifications 18-20 .
This model yields reproducible infarcts and increased post-survival rates. Approximately 80% of the ischemic strokes in humans happen in the MCA area 21 and thus this is a very relevant model for stroke studies. Currently, there is a paucity of effective treatments available to stroke patients, and thus there is a need for good models to test potential pharmacological compounds and evaluate physiological outcomes. This method can also be used for studying intracellular hypoxia response mechanisms in vivo. 
Here, we present the MCA ligation surgery in a C57/BL6 mouse. We describe the pre-surgical preparation, MCA ligation surgery and 2,3,5 Triphenyltetrazolium chloride (TTC) staining for quantification of infarct volumes.

Middle cerebral artery (MCA) ligation is a technique to study focal cerebral ischemia in animal models. In this method, the middle cerebral artery is exposed by craniotomy and ligated by cauterization. This method gives highly reproducible infarct volumes and increased post-operative survival rates compared to other methods available.

在小鼠永久性大脑中动脉结扎术中的应用

Focal cerebral ischemia is among the most common type of stroke seen in patients. Due to the clinical significance there has been a prolonged effort to ...

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 ...

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 ...

Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, obesity and diabetes. Accordingly, when evaluating therapies for stroke in animals, it is important to select a model with excellent face validity. Ischemic stroke accounts for 80% of all strokes, and the majority of these occur in the territory of the middle cerebral artery (MCA), often inducing infarcts that affect the sensorimotor cortex, causing persistent plegia or paresis on the contralateral side of the body. We demonstrate in this video a method for producing ischemic stroke in elderly rats, which causes sustained sensorimotor disability and substantial cortical infarcts. Specifically, we induce permanent distal middle cerebral artery occlusion (MCAO) in elderly female rats by using diathermy forceps to occlude a short segment of this artery. The carotid artery on the ipsilateral side to the lesion was then permanently occluded and the contralateral carotid artery was transiently occluded for 60 min. We measure the infarct size using structural T2-weighted magnetic resonance imaging (MRI) at 24 hr and 8 weeks after stroke. In this study, the mean infarct volume was 4.5% &#177; 2.0% (standard deviation) of the ipsilateral hemisphere at 24 hr (corrected for brain swelling using Gerriet&#8217;s equation, n = 5). This model is feasible and clinically relevant as it permits the induction of sustained sensorimotor deficits, which is important for the elucidation of pathophysiological mechanisms and novel treatments.

Here we present a protocol to produce permanent distal middle cerebral artery occlusion in elderly female rats with simultaneous occlusion of the carotid arteries to generate large cortical infarcts and sustained deficits. We show confirmation of the lesion size using structural MRI at 24 hr and 8 weeks after stroke.

执行永久远端大脑中与老年大鼠颈总动脉闭塞研究皮层缺血具有持续残疾

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, ...

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.