这项工作是由美国国立卫生研究院，加州大学旧金山分校REAC奖授予NS057195和肯特州立大学的创业基金，以WH周的支持。

<html> 1。缺血方法该协议被批准机构动物护理和使用委员会，加州大学旧金山分校和美国肯特州立大学，并遵守国家机构使用实验动物的健康指引。 <ol></ol><ol><li>切成20毫米段5-0单丝缝合（哈佛器械，Holliston，MA）。回合每个细分市场的一角，暖气附近cauterizer（布伦特里科学公司，布伦特里，马）。测量的提示使用千分尺（应用图像公司，罗彻斯特，纽约州）的直径。我们使用缝合最后的技巧鼠标与0.21-0.22毫米直径机身重量为25-30克。 </li><li>所有的手术工具消毒高压灭菌（最低121℃，15 Psi范围，为15分钟，）。使用70％乙醇消毒手术表及相关设备。 </li><li>麻醉一个8-12周龄小鼠（25-30克）用5％的异氟醚（Aerrane，巴克斯特，迪尔菲尔德，IL）70％，30％O 2 / N 2 O的（VetEquip，公司使用的V - 10麻醉系统，普莱森顿，CA）。麻醉诱导后，降低异氟醚的水平，并维持在1.5％。 </li><li>鼠标放置在加热垫的仰卧姿势。插入直肠探头，监控并保持身体温度之间36.5-37.5℃使用的TR - 200 homeothermic温度系统（精细的科学工具公司，福斯特市，CA）。 </li><li>剃除电动快船（布伦特里科学）的皮肤暴露的腹侧颈部的皮毛。使用三个应用70％乙醇消毒手术部位。 </li><li>在一个立体解剖显微镜（日本尼康公司，），1厘米长的颈部正中切口。使用拉钩（布伦特里科学）暴露手术野，并确定右侧颈总动脉（CCA），颈外动脉（ECA）和颈内动脉（ICA）。仔细解剖动脉周围神经及筋膜。 </li><li>进一步远端解剖非洲经委会和凝聚非洲经委会和甲状腺上动脉（STA）的分支，使用双极电凝（霍华德仪器公司，塔斯卡卢萨，铝）。在凝结段切非洲经委会和STA。 </li><li>松散地挂在非洲经委会残端两个8-0丝缝合。应用到非洲经委会和ICA的CCA分叉血管钳（精细科学的工具）。 </li><li>设为Vannas式弹簧剪刀（精细科学的工具），在非洲经委会残端年底的一个小切口。尖圆5-0单丝缝合的长度测量和记录。插入切口缝合，并提前钳。收紧两个丝线缝合管腔刚好够安全还保留在住宅单丝缝合的流动性。 </li><li>取出钳从分叉。轻轻提前进入ICA单丝缝合了9-10毫米以外的CCA分叉处闭塞马华起源的距离，从非洲经委会流明。手术时间约30-45分钟。 </li><li>缝合颈部切口和鼠标放置在一个35℃护理框从麻醉中恢复，并返回它的笼子。通常需要5-10分钟的小鼠从麻醉中恢复。要执行的瞬态缺血，研究人员可以重新麻醉鼠标，并撤回到非洲经委会残端缝合后一段时间，通常在0.5-2小时</li><li>缺血诱导的二十四小时后，用5％的异氟醚麻醉的鼠标和安乐死颈椎脱位。杀头的鼠标和收集的大脑。 2毫米的大脑矩阵冰（布伦特里科学）分为四个片，脑冠状切片。孵育在2％的大脑切片2,3,5 - 三苯基氯化四氮唑（TTC）（Sigma - Aldrich公司）在1X PBS在室温为20分钟，以确定梗死的大小和程度。修复10％的脑切片中性缓冲福尔马林溶液（Sigma - Aldrich公司）在4 ° C，直到成像。在朱庇特协议955（ <a href="http://www.jove.com/index/Details.stp?ID=955">http://www.jove.com/index/Details.stp?ID=955</a> ）9梗死的程度是可以量化的。 </li></ol> 2。代表性的成果<ol></ol>在纹状体和背外侧皮层缺血产生的梗塞。纹状体是大脑皮层缺血更为敏感。在缺血30分钟会产生仅在纹状体梗塞，而超过一个小时的缺血损伤纹状体和皮层。永久性缺血24 h后，总梗塞百分比〜40 ±半球和手术后的死亡率5％〜10％。我们排除进一步研究老鼠如果在手术过程中发生出血过多，手术时间超过90分钟，小鼠从麻醉在15分钟内未能收回，或出血是在大脑切片或Willis环的基地在尸检发现。 jove_content“&gt; <img src="/files/ftp_upload/2761/2761fig1.jpg" alt="图1" /> TTC染色脑切片图1。代表性的图像（冠状1-4级）永久性缺血24小时后。在活组织TTC酶减少脱氢酶1,3,5 triphenylformazan（TPF），这是红色的，而在坏死区，它仍然是白色的，由于缺乏这种酶的活性。因此，梗死面积，可以认定其白色由于缺乏转换的TTC TPF。注：TTC是有点热和光不稳定，以便从尽可能多的热和光保护染色切片。

<ol>
	<li>Howard, G., Howard, V. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Distribution of stroke: hetrogeneity of stroke by age, and sex. , 3-12 (2004).</li><li>Thom, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+disease+and+stroke+statistics--2006+update:+a+report+from+the+American+Heart+Association+Statistics+Committee+and+Stroke+Statistics+Subcommittee.">Heart disease and stroke statistics--2006 update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee.</a> Circulation. 113, e85-e151 (2006).</li><li>Mohr, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Middle cerebral artery disease. , 123-166 (2004).</li><li>Longa, E. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reversible+middle+cerebral+artery+occlusion+without+craniectomy+in+rats.">Reversible middle cerebral artery occlusion without craniectomy in rats.</a> Stroke. 20, 84-91 (1989).</li><li>Chou, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neutrophil+protein+kinase+Cdelta+as+a+mediator+of+stroke-reperfusion+injury.">Neutrophil protein kinase Cdelta as a mediator of stroke-reperfusion injury.</a> J. Clin. Invest. 114, 49-56 (2004).</li><li>Hudgins, W. R., Garcia, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+electrocautery,+atmospheric+exposure,+and+surgical+retraction+on+the+permeability+of+the+blood-brain-barrier.">The effect of electrocautery, atmospheric exposure, and surgical retraction on the permeability of the blood-brain-barrier.</a> Stroke. 1, 375-380 (1970).</li><li>Carmichael, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+of+focal+stroke:+size,+mechanism,+and+purpose.">Rodent models of focal stroke: size, mechanism, and purpose.</a> NeuroRx. 2, 396-409 (2005).</li><li>Durukan, A., Tatlisumak, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+ischemic+stroke:+overview+of+major+experimental+rodent+models,+pathophysiology,+and+therapy+of+focal+cerebral+ischemia.">Acute ischemic stroke: overview of major experimental rodent models, pathophysiology, and therapy of focal cerebral ischemia.</a> Pharmacol. Biochem. Behav. 87, 179-197 (2007).</li><li>Taniguchi, H., Andreasson, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hypoxic-ischemic+encephalopathy+model+of+perinatal+ischemia.">The hypoxic-ischemic encephalopathy model of perinatal ischemia.</a> J Vis Exp. , (2008).</li></ol>

我们没有透露任何潜在的利益冲突。

在小鼠缺血是常用来模拟人类的脑缺血。中风研究小鼠的使用越来越频繁，由于转基因和基因敲除株的可用性。有几个重要的细节要注意在协议： <ol><li>重要的是在手术过程中保持体温鼠标，和之前完全从麻醉中复苏。体温已梗死的程度上的影响;低温跌幅和热疗增加梗死面积。 </li><li>在揭露和孤立CCA和非洲经委会的，避免破坏附近的迷走神经及气管，这可能会增加梗死面积，减少的可行性。 </li><li>切勿将超过10毫米的单丝缝合通过分叉。太远插入一个缝合穿孔导致脑出血，大脑前动脉。超出了分叉点约9-10毫米先进的缝合时，可以感觉到阻力。如果发生这种情况，研究人员应停止前进的缝合和确认距离。 </li></ol>

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		<div>
		<table border="1">
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		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>Isoflurane</td>
<td>Chemical	</td>
<td>Aerrane, Baxter</td>
<td>95045-588</td>
<td>&nbsp;</td>
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<td>V-10 Anesthesia system</td>
<td>Equipment</td>
<td>VetEquip</td>
<td>901807</td>
<td>&nbsp;</td>
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<td>TR-200 Temp Controller</td>
<td>Equipment</td>
<td>Fine Science Tools</td>
<td>21060</td>
<td>&nbsp;</td>
<tr>
<td>Electric clipper</td>
<td>Equipment</td>
<td>Braintree</td>
<td>CLP-9931</td>
<td>&nbsp;</td>
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<td>Dissecting microscope</td>
<td>Equipment</td>
<td>Nikon</td>
<td>SMZ745T</td>
<td>&nbsp;</td>
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<td>Retractor system</td>
<td>Equipment</td>
<td>Braintree</td>
<td>ACD-014</td>
<td>&nbsp;</td>
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<td>Bipolar coagulator</td>
<td>Equipment</td>
<td>Howard Instrument</td>
<td>64000</td>
<td>&nbsp;</td>
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<td>Silk suture</td>
<td>Material</td>
<td>Harvard Apparatus</td>
<td>510479</td>
<td>&nbsp;</td>
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<td>Monofilament suture</td>
<td>Material</td>
<td>Harvard Apparatus</td>
<td>723351</td>
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<td>Vascular clamp</td>
<td>Equipment</td>
<td>Fine Science Tools</td>
<td>00396-01</td>
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<td>Vannas scissor</td>
<td>Equipment</td>
<td>Fine Science Tools</td>
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<td>10% neutral buffer formalin</td>
<td>Chemical</td>
<td>Sigma-Aldrich</td>
<td>HT5011</td>
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mouse model of middle cerebral artery occlusion

中风是最常见的致命的神经系统疾病在美国的1 。大多数的中风导致脑血管堵塞（缺血性中风）2（88％）。由于在大脑中动脉（MCA）3境内发生的最缺血性中风（〜80％），已开发的许多动物中风 ​​车型都集中在此动脉。腔内单丝大脑中动脉闭塞（MCAO）模型涉及到颈外动脉手术长丝插入线程到颈内动脉（ICA），直到提示阻塞的MCA的起源，在停止血流量和随后的脑梗死MCA 领土 4 。该技术可用于模型永久或暂时闭塞5的。如果删除后一定的时间间隔（30分钟，1小时或2小时）缝合，是实现再灌注（瞬态缺血）;如果灯丝是留在原地（24小时）的程序是适合作为一个永久性缺血模型。这种技术并不需要去骨瓣，神经外科手术过程来删除一个头骨的一部分，这可能会影响颅内压和温度 6 。它已成为最常用的方法来模仿永久性和短暂局灶性脑缺血大鼠和小鼠7,8。为了评估脑梗死的严重程度，我们与2,3,5 -三苯基氯化四氮唑（TTC）染色脑切片，以确定脑组织缺血9。在这段视频中，我们表现出缺血的方法和TTC染色测定梗死面积。

Watch this Scientific Journal Video about Mouse Model of Middle Cerebral Artery Occlusion at JoVE.com

Mouse Model of Middle Cerebral Artery Occlusion

在视频中，我们证明了生产中使用腔内单丝的成年小鼠的大脑中动脉闭塞的方法。我们还展示了如何评价脑梗死的严重程度，由2,3,5 - 三苯基氯化四氮唑（TTC）染色。

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 ...

mouse-model-of-middle-cerebral-artery-occlusion

Research

JoVE Journal

Medicine

77.3K 次观看.  Ernest Gallo Clinic and Research Center, University of California, San Francisco. 在视频中，我们证明了生产中使用腔内单丝的成年小鼠的大脑中动脉闭塞的方法。我们还展示了如何评价脑梗死的严重程度，由2,3,5 - 三苯基氯化四氮唑（TTC）染色。

视频: Mouse Model of 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

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 ...

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 ...

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 ...

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week protocol for repetitive hypoxic preconditioning (RHP) induces long-term protection against central nervous system (CNS) injury in a mouse model of focal ischemic stroke. RHP consists of 9 stochastic exposures to hypoxia that vary in both duration (2 or 4 hr) and intensity (8% and 11% O2). RHP reduces infarct volumes, blood-brain barrier (BBB) disruption, and the post-stroke inflammatory response for weeks following the last exposure to hypoxia, suggesting a long-term induction of an endogenous CNS-protective phenotype. The methodology for the dual quantification of infarct volume and BBB disruption is effective in assessing neurovascular protection in mice with RHP or other putative neuroprotectants. Adult male Swiss Webster mice were preconditioned by RHP or duration-equivalent exposures to 21% O2 (i.e. room air). A 60 min transient middle cerebral artery occlusion (tMCAo) was induced 2 weeks following the last hypoxic exposure. Both the occlusion and reperfusion were confirmed by transcranial laser Doppler flowmetry. Twenty-two hr after reperfusion, Evans Blue (EB) was intravenously administered through a tail vein injection. 2 hr later, animals were sacrificed by isoflurane overdose and brain sections were stained with 2,3,5- triphenyltetrazolium chloride (TTC). Infarcts volumes were then quantified. Next, EB was extracted from the tissue over 48 hr to determine BBB disruption after tMCAo. In summary, RHP is a simple protocol that can be replicated, with minimal cost, to induce long-term endogenous neurovascular protection from stroke injury in mice, with the translational potential for other CNS-based and systemic pro-inflammatory disease states.

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

神经血管保护定量继在小鼠重复缺氧预处理和瞬态大脑中动脉闭塞

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week ...

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 ...

Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature and mature brain. Stroke occurs commonly in the perinatal period in humans, and transient ischemia-reperfusion is the most common form of stroke in neonates. The reperfusion phase is a critical component of injury progression, which occurs over a period of days to weeks, and of the endogenous response to injury. This postnatal day 10 (p10) rat model of transient middle cerebral artery occlusion (tMCAO) creates a unilateral, non-hemorrhagic focal ischemia-reperfusion injury that can be utilized to study the mechanisms of focal injury and repair in the full-term-equivalent brain. The injury pattern that is produced by tMCAO is consistent and highly reproducible and can be confirmed with MRI or histological analyses. The severity of injury can be manipulated through changes in occlusion time and other methods that will be discussed.

Neonatal stroke is a significant cause of early brain injury requiring a translational model with consistent focal injury patterns and high reproducibility in order to enable study. This study describes the detailed surgical procedure for creating a non-hemorrhagic, unilateral focal ischemia-reperfusion injury in full-term-equivalent rodents.

在P10大鼠新生儿中风的短暂大脑中动脉栓塞模型

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature ...