Name: permanent cerebral vessel occlusion via double ligature and transection
Uploaded: 2013-07-21T14:00:00
Description: Watch this Scientific Journal Video about Permanent Cerebral Vessel Occlusion via Double Ligature and Transection at JoVE.com

支持这项工作是由美国心脏协会的博士前奖学金788808-41910，NIH-NINDS NS-066001和NS-055832，该中心的听力研究NIH训练1T32DC010775，-01。

1。入门：需手术器械参见图4 <ol><li>牙钻（卡瓦牙科设备，型号：UMXL-TM），2位钻头，钻和3位</li><li> 〜30号注射针头</li><li>镊子，弯尖锯齿可选（可以是有益的，但​​不是必需的） </li><li>两个精尖镊子</li><li>钢丝钳</li><li>缝合线</li><li>微型剪刀</li></ol> 2。创建外科窗口<ol><li>麻醉：程序是在遵守与美国国立卫生研究院的指导方针，并已通过...

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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+critical+reevaluation+of+the+intraluminal+thread+model+of+focal+cerebral+ischemia:+evidence+of+inadvertent+premature+reperfusion+and+subarachnoid+hemorrhage+in+rats+by+laser-Doppler+flowmetry.">A critical reevaluation of the intraluminal thread model of focal cerebral ischemia: evidence of inadvertent premature reperfusion and subarachnoid hemorrhage in rats by laser-Doppler flowmetry.</a> Stroke. 29, 2162-2170 (1998).</li></ol>

此协议是为了诱导缺血啮齿动物皮质内，这样做实验对象用最少的外围影响。双闭塞和横断法允许的视觉确认该船只已被永久封闭，并没有过多的入侵或组织损伤，可以进行，成活率高。此闭塞协议可能被应用到任何皮质船只可通过开颅手术，以促使在一个特定的域皮质缺血。此外，这样的闭塞时，可以执行一个动物是在立体定位仪上，允许同时使用各种研究的技术，如功能性成像或电生理记录?...

为了有效地模仿人类缺血性中风的诱发缺血性条件，广泛采用的几种动物卒中模型，不同卷梗死。在光化学模型中，脑是通过完整的头骨照射的光敏物质（如孟加拉玫瑰）静脉注射后，用激光照射导致光化学凝血的照射血管堵塞，缺血周围组织6内， 7。 photothrombosis可以导致在非常小的，孤立的梗死区域，通常被用作一种手段，建模“小冲程”，或“微招。 更广泛采用的技术诱导缺血性中风，尤其是在大脑中动脉（MCA），所述管腔内，灯丝手术引入颈外动脉的前端前进，直到阻塞MCA的基础上的单丝模型8。 PRI的线栓闭塞玛丽的挑战是高死亡率（70％MCA闭塞时3小时，有关中风研究的时间点）9。其他问题的方法包括可能的蛛网膜下腔出血，不完全闭塞，可变梗死体积10,11。这种模式的结果梗死的广泛程度在皮层和皮层下12，和一个巨大的人中风的模型。 虽然微观和严重的中风模型是重要的，但人类中风通常是介于两者之间。在大型临床研究，中风梗塞范围的大小从28-80厘米3，转换为4.5-14％，局部缺血性同侧半球9。相比较而言，我们的大鼠PMCAO梗死面积范围从约9-35毫米3，构成3〜12％的同侧局部缺血性大脑半球。我们的PMCAO模型，因此，类似于人类缺血性中风梗塞体积百分比脑音量。 除了造型结构损伤人的条件相似的功能和行为缺陷的的中风，PMCAO结果。至少，中风导致运动障碍，对侧13-15中风损伤，损失或中断的运动和感觉功能16,17，诱发神经元的活动16,18损失或中断，脑血流量减少19的有效模式， 20，梗死21,22。因此，我们PMCAO的模型严重的MCA闭塞导致身体残疾，丧失功能在感觉皮层（及邻近皮层），神经元活动的中断，严重的大脑中动脉的血流量减少，梗死标志属性缺血性中风23 -25，因此作为人类中风的有效模式。 在程序上，PMCAO涉及小的开颅手术中，我们小心地取出颅骨和硬脑膜2×2毫米的“外科手术式窗口”，只是在初始段（M1）马华前马华前部和后部皮质分支（ 图1A和1B）的主要分歧。我们通过半曲线反向切割缝合针和线（6-0丝绸），通过软脑膜层的脑膜，低于MCA和上述皮层的表面上（见表必要的外科用品的特定的试剂和设备进行PMCAO ）。我们再配合双重结扎，收紧马华两节左右，横切两节之间的船只。发生的双重结扎横断通过M1恰好远侧豆纹支化的，例如，皮质分支MCA的影响，从而发生皮质梗塞（无皮质下损伤）26,27（ 图2）。虽然人类卒中往往涉及到皮质下梗死，造型在啮齿类动物中，需要增加的侵袭（阻塞脑血管皮质branchi之前吴先生需要访问通过在脖子上的颈动脉的动脉，必须在技术和梗死面积的变化增加额外闭塞）。这里所描述的模型，不能执行更近侧的早期分支MCA 通过一个简单的开颅手术是不可能的。虽然它可能是手术可能诱发皮层下通过PMCAO梗死，闭塞将需要一个非常侵入性的程序，因此并不理想。 闭塞效果，可以确认通过激光多普勒激光散斑成像12,24,25（ 图3），或组织学宰后（ 图2）。应该指出的是，以往的研究表明，感官刺激可以起到主要作用于心肌梗死的演变和结果赋予保护免受损害时2小时内PMCAO，造成给药后3小时PMCAO上增加中风损伤时24,25,28。我们已经证实，在5小时后PMCAO，刺激不再有效果的结果（未发表资料）。因此，感官刺激的主题应该被最小化后5小时，获得PMCAO梗死体积最小变异。因此，本集团运行“未处理”这种类型的控制，保持麻醉大鼠5小时后PMCAO的，在黑暗中，用最少的感官刺激，并明确表示无晶须的刺激。 还应当指出的是，偶尔在MCA的结构变化，包括过度支化，多个主段，或没有交通动脉可以发生在频率为10〜30％的雄性成年Sprague Dawley大鼠29,30。如果马华异常的观察，最好是不使用特定的主题添加动物如血管异常，会增加心肌梗死的变异。 此外，还有一些实用方面的o你的程序，使这个闭塞的方法，便于中风调查。首先，缝合线可能被放置周围的动脉，但没有拧紧以收集基线评估，然后结扎横断后缺血后评估，。以这种方式，有效地控制了手术的准备所需的闭塞，在主体之内。由于主体可能会保持静止或整个闭塞的一个立体框架内，就可以进行各学科的实验评估之前，期间和之后，闭塞而移动主题或干扰任何实验设备在使用25,28。此外，这一过程的结果在一个非常低的死亡率，即使在老化啮齿动物受试者21-24个月的婴儿（相当于老年人类）31，并且因此可以用于评估的影响，中风的治疗方法更精确地模拟最常见的中风患者的年龄段25,28。船只transectio的N还提供一些实用的目的。横断后出血的情况下确认该船只都结扎站点完全闭塞。此外，横断确保一个永久的血流中断。最后，切断确保闭塞血管的前端部的任何检测到的血流量必须来自备用源。 最后，虽然具体描述此技术MCA闭塞在这个手稿和视频，相同的双重结扎横断技术可能适用于任何可通过开颅脑血管。例如，我们的实验室，利用PMCAO配合一些额外的永久闭塞远端MCA分支，以阻止小学和侧支血流24的方式，类似的技术设计选择性诱导缺血内的初级躯体感觉皮层32。 总之，T他的方法适用于马华永久闭塞密切模拟人类缺血性中风的三个主要方面：最常见的位置（MCA），类型（缺血），并与人类中风的临床文献相关程度的损伤（心肌梗死）。此外，闭塞该方法可被应用到整个大脑的单个或多个闭塞站点，也可以进行具有高的存活率老年受试者。鉴于动态的，永久性的，相对无创性闭塞，这种技术代表评价新方法，为保护和治疗中风的临床前研究人员一个额外的工具。

<table border="1">
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 <td>Name of the equipment</td>
 <td>Company</td>
 <td>Catalogue number</td>
 <td>Comments (optional)</td>
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 <td>Extra Fine Graefe Forceps - 0.5 mm Tips Slight Curve (1)</td>
 <td>Fine Science Tools </td>
 <td>11151-10</td>
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 <td>Ceramic Coated Dumont #5 Forceps (2)</td>
 <td>Fine Science Tools </td>
 <td>11252-50</td>
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 <td>Extra Fine Bonn Scissors, straight (1)</td>
 <td>Fine Science Tools </td>
 <td>14084-08</td>
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 <td>Round 3/8 (16 mm) Suture Needles</td>
 <td>Fine Science Tools</td>
 <td>12050-02</td>
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 <td>6-0 Braided Silk Suture</td>
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 <td>NC9071061 
 Harvard Apparatus 
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 <td>30 gauge needle, &frac12;" length</td>
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</table>

permanent cerebral vessel occlusion via double ligature and transection

中风是导致死亡，残疾和全球社会经济损失。大多数中风导致血流中断（缺血）1。大脑中动脉（MCA）提供了一个伟大的大部分血液皮层2的侧面，是人类中风的最常见的部位，并在其领土内的缺血会导致广泛的功能障碍或死亡1,4,5。缺血性中风的幸存者经常遭受损失或中断的机动能力，感觉障碍，心肌梗死。在努力捕捉到这些中风的关键特性，从而制定有效的治疗方法，一个很大的重点是放在MCA缺血动物模型。 在这里，我们提出了一个永久阻断皮质表面血管的方法。我们将这种方法使用一个例子，相关的血管闭塞，模拟最常见的类型，位置和OUTCOM辰人力中风，永久性大脑中动脉闭塞（PMCAO）。在这个模型中，我们手术暴露马华在成年大鼠，通过双重结扎和切断船只随后闭塞。这PMCAO阻止皮质支近端马华MCA皮质境内，大部分的皮质，造成缺血。 ，闭塞此方法也可用于皮质血管闭塞更远侧的部分，以实现局灶性脑缺血，针对一个较小的皮层区域。的PMCAO的主要缺点是，有些侵入性外科手术作为一个小的开颅手术需要访问MCA，虽然这样的结果在最小的组织损伤。然而，这种模式的主要优点是：定义，血流量减少的程度是一致的，功能和神经功能缺损发生迅速闭塞部位，梗死面积是一致的，并且可以长期生存率高的长期慢性评估。

可以确认使用的激光散斑成像（LSI），其中包括血流成像技术成功闭塞的船只。 MCA主要的皮质分支的血流量下降到〜25％的基准以下的闭塞，根据在记录系统中的噪声和灵敏度的技术水平。见如图3段的皮质支马华的代表LSI形象前后的大脑中动脉闭塞。当所描述的闭塞技术应用到MCA M1段，阻止所有皮质马华分支机构，并防止〜5小时以下闭塞的感官刺激，结果是28.4±2.4毫米?...

Watch this Scientific Journal Video about Permanent Cerebral Vessel Occlusion via Double Ligature and Transection at JoVE.com

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

我们描述了一个高度可重复性的方法永久啮齿类动物的主要脑血管闭塞。这种技术可以完成与周损伤非常少，最小的失血，长期生存的一种高速率，和一致的梗死体积与人类的临床人口。

永久性脑血管闭塞<em&gt;通过</em&gt;双人结扎横断

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

Research

JoVE Journal

Medicine

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

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

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

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 ...

The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

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 ...

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

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 ...

2-Vessel Occlusion/Hypotension: A Rat Model of Global Brain Ischemia

2血管闭塞/低血压：全脑缺血模型大鼠

Cardiac arrest followed by resuscitation often results in dramatic brain damage caused by ischemia and subsequent reperfusion of the brain. Global brain ...

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

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

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

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 ...

Stereological and Flow Cytometry Characterization of Leukocyte Subpopulations in Models of Transient or Permanent Cerebral Ischemia

白细胞亚群在暂时或永久的脑缺血模型体视学和流式细胞仪鉴定

Microglia activation, as well as extravasation of haematogenous macrophages and neutrophils, is believed to play a pivotal role in brain injury after ...

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

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 ...