This study was supported by the NIH grant NS074559 (to C. K.). We thank all collaborators who contributed to our research articles that the present methodology report is based upon.

该协议是由埃默里大学的机构动物护理和使用委员会（IACUC）批准，并遵循健康指导全国学院的护理和使用实验动物。 1.安装<ol><li>准备手术创面上变暖垫与热泵连接，在37℃下在手术前至少15分钟。放置使用3毫升注射器针筒上的手术床上的颈部辊。准备麻醉气体，在医用空气2％异氟醚。 </li><li>准备蒸压钳，剪刀，针微型持有，止血，棉签和缝线。准备组织胶和眼膏。 </li><li>建立缺氧系统和温度控制器，加热灯泡和直肠探头。准备缺氧的气体与7.5％O 2 92.5％N 2平衡2％异氟醚。 </li><li>前一小时的手术，将小鼠通过皮下注射缓释美洛昔康（4.0毫克/千克）的analgesized。 </li></ol> &lt;P类="j​​ove_title"&gt; 2。短暂性脑缺氧缺血（图1B） <ol><li>麻醉10-13周龄雄性C57BL / 6小鼠称重22至30克的麻醉诱导室用3％异氟烷，直到动物没有反应到脚挤压，然后使用电子剃须刀卸下右颈部的头发。 </li><li>放置老鼠在手术床上以2升/分钟的流速，用2％异氟烷连接在医用空气。安全前肢伸出脖子上沿辊使用医用胶带两侧。 </li><li>清洁手术部位的切口优碘其次是酒精，然后棉签。 </li><li>在解剖显微镜，使用直镊子和剪刀微型约0.2厘米的皮肤中线做横向将0.5cm右颈部切口。 </li><li>使用一对细锯齿钳子拉开筋膜和组织以暴露右颈总动脉（RCCA）。小心地用一双细镊子流畅的迷走神经分离RCCA。 </li><li>在线结2预切割的5-0丝线缝合（释放的）上的RCCA，然后缝合用4-0尼龙单丝缝合（ 图1C）的皮肤。 </li><li>适用于双眼眼药膏，以防止干燥。 </li><li>小鼠迅速转移到低氧系统，把鼻子和嘴中的面罩，用2％异氟烷在7.5％ 的 O 2在0.5-1，流量率/分钟进行30分钟。 <ol><li>在缺氧，使用温度控制器，加热灯控制肛温在37.5±0.5°C。监测呼吸率在80-120次/ min。的体温高于37℃，维持在缺氧时重要的是要创建一致脑梗塞。低呼吸频率通常在20分钟缺氧发生。取下面罩，允许正常供气若跌破呼吸频率下降到40。这需要1-2分钟，不计算入30分钟缺氧时间。 </li></ol></li><li>缺氧后，乘车小鼠到外科床和从RCCA释放这两个缝线。收使用组织胶伤口，然后返回小鼠的笼中。排除动物如果同时满足两个活节缺氧后意外释放。 </li><li>监测小鼠为5-10分钟，以从缺氧和麻醉中恢复。将湿的食物，在笼子里，并返回到动物护理设施。 注:THI动物展示后在24小时轻度到重度盘旋的行为是相关脑违规行为。大多数动物癫痫症状THI后的24小时时间点之前死亡。 </li></ol> 3.激光散斑对比度成像注意:虽然这不是THI模式的基本过程，二维激光散斑对比度成像系统16可以被用于在或者短暂缺氧缺血后表征脑血流量（CBF）的变化。到ST后立即记录CBF的变化THI下，记录EP 2.6。另外，以THI侮辱后的比较CBF的恢复，这些程序可以按照步骤2.10进行。 <ol><li>将麻醉小鼠俯卧位，并与外露，但在未开封的头骨进行头皮1厘米长的正中切口。 </li><li>监测CBF在两个大脑半球下根据制造商的协议的血流成像器并启动CCAO手术（步骤2.6）后，立即记录脑血流量。持续50分钟。 </li><li>显示CBF图像与任意单位在16色调色板和实时分析使用MoorFLPI软件按照制造商的说明（ 图2）的选择区域。 </li><li>记录CBF图像后，关闭与组织胶头皮和动物回到笼子里。 </li></ol> 4. tPA的管理<ol><li>注入动物尾部静脉与溶剂或10mg / kg的重组的tPA（220-300＆＃956;微升1毫克/米的tPA）在0.5，1或tCCAo加缺氧后4小时（ 图4）。 </li></ol> 5.脑损伤检测几种不同的选项注意:为了收集脑组织样本，安乐死的小鼠THI后1,4或24小时。 <ol><li>通过体内 2,3,5-三苯基氯化执行梗塞体积的定量（TTC）方法，在24小时THI侮辱为先前的描述后，17 <ol><li>腹膜内注射的动物用1.4M的甘露醇溶液（±0.1毫升/克体重）穿心灌注前30分钟。穿心灌流小鼠用PBS，接着用10毫升2-％TTC的。 </li><li> 10分钟后除去动物手术器械的大脑并放入4％多聚甲醛固定过夜，并部分成1mm厚度与vibratome。 </li><li>数码显微镜和定量捕捉了四大系列切片脑幻灯片IFY梗死体积为梗塞区（白色区域中的右侧），以使用ImageJ的软件的损坏，对侧半球的面积之比。 </li></ol></li><li>另外，通过免疫荧光THI在损伤后1小时进行血栓形成。 <ol><li>冻结华侨城化合物和部分使用恒温器的大脑在12微米厚的固定大脑。 </li><li>孵育兔抗血纤蛋白原抗体大脑滑动（1:100），按照由山羊抗兔Alexa的FLURO 488染料（1:200），观察荧光在荧光显微镜上。 </li></ol></li><li>或者，通过尾静脉注射100微升2％伊文思蓝染料在THI损伤后4小时进行血管阻塞。 <ol><li>安乐死的小鼠，迅速地砍掉头埃文斯蓝注射后去除大脑到4％多聚甲醛。注意:这需要5-10分钟的伊文思蓝流通与两个fore-的蓝色和后肢。 </li><li> Secti上固定的大脑，在100微米厚使用滑动切片机，并使用680 nm发射滤波器上的荧光显微镜观察荧光。 </li></ol></li></ol>

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中风是不断增长的意义随着人口老龄化社会的任何一个主要的健康问题。从全球来看，中风是死亡的第二大原因与2010年估计590万死亡事件，相当于所有的死亡病例18 11.1％。脑卒中也是伤残调整生命年第三主要原因（伤残调整寿命）在2010年失去了全球，从第五位，1990年上升19这些流行病学数据突出的急性（缺血性）中风的更有效的治疗需要。然而，尽管大量研究在临床前的神经保...

溶栓再通的急性缺血性中风在临床实践中1最有效的疗法。然而，大多数的临床前神经保护研究的在瞬态机械梗阻模型（线栓大脑中动脉闭塞），产生在去除血管闭塞的脑血流的快速恢复，并显示很少或几乎没有益处由tPA的溶栓进行。它已被提出，中风模型的可疑的选择作出了贡献，至少部分地难度在翻译神经保护治疗的病人2,3。因此，有越来越多的要求使用tPA的响应血栓栓塞性中风模型在临床前研究，但这种模式也有技术问题（见讨论）4-7。在这里，我们描述了基于单边短暂缺氧缺血（THI）侮辱，其反应静脉注射tPA的治疗8一种新的血栓性中风模型。 基于该列文过程（单侧颈总动脉，随后通过暴露于短暂缺氧在一个腔室的永久的结扎后），这是发明了用于与成年大鼠实验在1960年9被开发的THI中风模型。原始的列文过程褪成朦胧因为它只产生可变的脑损伤，但同样的侮辱在啮齿动物的幼崽引起神经病理学一致，当它被重新引入由罗伯特·万努奇和他的同事们为新生儿缺氧缺血性脑病（HIE）1981年10模型。近年来，一些研究者通过在低氧室11调节温度重新调整了莱文-万努奇模型成年小鼠。它是合理的，不一致的脑部病变原莱文过程中可能出现的在缺氧室波动体温成年老鼠的。为了检验这一假设，我们修改了莱文的程序通过给予低氧气体通过面罩，同时保持啮齿类核心温度在37℃下在手术台12上 。正如预期的那样，严格的体温控制大大提高了高新所致的脑病变可重复性。该HI损伤也触发凝血，自噬，和灰度和白质损伤13。其他研究者也采用了HI模型，研究脑卒中后炎症反应14。 在HI中风模型的独特之处在于它紧随魏尔啸的黑社会的血栓形成，其中包括血流量，内皮损伤（ 如由于HI-诱导的氧化应激），和高凝状态（HI-诱导的血小板活化）的瘀（ 图1A）15。正因为如此，对HI模型可以捕获有关真实世界缺血性中风的一些病理生理机制。有了这个念头在脑海中，我们进一步完善了HI模型与联合国的可逆性结扎ilateral颈总动脉（因此创建一个短暂的HI侮辱），并测试其反应的tPA溶栓或无依达拉奉。依达拉奉是已经批准在日本范围内发生9小时24治疗缺血性中风自由基清除剂。我们的实验表明，简短的30分钟短暂的HI引发血栓性脑梗死，并结合tPA的，依达拉奉治疗赋予协同效益8。在这里，我们描述了详细的外科手术和THI模型的方法上的考虑，它可以被用来优化急性缺血性中风的再灌注治疗。

<table><tbody><tr><td>adult male mice</td><td>Charles River</td><td>C57BL/6&amp;nbsp;</td><td>10-13 weeks old (22-30 g)</td></tr><tr><td>Mobile Laboratory Animal Anesthesia System</td><td>VetEquip</td><td>901807</td><td>anesthesia</td></tr><tr><td>Medical air (Compressed) air tank</td><td>Airgas</td><td>UN1002</td><td>anesthesia</td></tr><tr><td>Isoflurane</td><td>Piramal Healthcare</td><td>NDC 66794-013-25</td><td>anesthesia</td></tr><tr><td>Multi-Station Lab Animal AnesthesiaSystem</td><td>Surgivet</td><td>V703501</td><td>hypoxia system</td></tr><tr><td>7.5% O&lt;sub&gt;2&lt;/sub&gt; balanced by 92.5% N&lt;sub&gt;2&lt;/sub&gt; tank</td><td>Airgas</td><td>UN1956</td><td>hypoxia system</td></tr><tr><td>Temperature Controller with heating lamp&amp;nbsp;</td><td>Cole Parmer&amp;nbsp;</td><td>EW-89000-10</td><td>temperature controllers</td></tr><tr><td>Rectal probe</td><td>Cole Parmer&amp;nbsp;</td><td>NCI-00141PG</td><td>temperature controllers</td></tr><tr><td>Dissecting microscope&amp;nbsp;</td><td>Olympus&amp;nbsp;</td><td>SZ40</td><td>surgical setup</td></tr><tr><td>Heat pump with warming pad</td><td>Gaymar&amp;nbsp;</td><td>TP700</td><td>surgical setup</td></tr><tr><td>Fine curved forceps (serrated)</td><td>FST</td><td>11370-31</td><td>surgical instrument</td></tr><tr><td>Fine curved forceps (smooth)</td><td>FST</td><td>11373-12</td><td>surgical instrument</td></tr><tr><td>micro scissors</td><td>FST</td><td>15000-03</td><td>surgical instrument</td></tr><tr><td>micro needle holders</td><td>FST</td><td>12060-01</td><td>surgical instrument</td></tr><tr><td>Halsted-Mosquito hemostats</td><td>FST</td><td>13008-12</td><td>surgical instrument</td></tr><tr><td>5-0 silk suture&amp;nbsp;</td><td>Harvard Apparatus</td><td>624143</td><td>surgical supplies</td></tr><tr><td>4-0 Nylon monofilament suture</td><td>LOOK</td><td>766B</td><td>surgical supplies</td></tr><tr><td>Tissue glue</td><td>Abbott Laboratories</td><td>NC9855218</td><td>surgical supplies</td></tr><tr><td>Puralube Vet ointment</td><td>Fisher</td><td>NC0138063&amp;nbsp;</td><td>eye dryness prevention&amp;nbsp;</td></tr><tr><td>MoorFLPI-2 blood flow imager</td><td>Moor</td><td>780-nm laser source</td><td>Laser Speckle Contrast Imaging</td></tr><tr><td>Mannitol</td><td>Sigma</td><td>M4125</td><td>&lt;i&gt;in vivo&lt;/i&gt; TTC</td></tr><tr><td>2,3,5-triphenyltetrazolium chloride (TTC)&amp;nbsp;</td><td>Sigma</td><td>T8877</td><td>&lt;i&gt;in vivo&lt;/i&gt; TTC</td></tr><tr><td>Vibratome</td><td>Stoelting</td><td>51425</td><td>brain section for &lt;i&gt;in vivo&lt;/i&gt; TTC&amp;nbsp;</td></tr><tr><td>Digital microscope</td><td>Dino-Lite</td><td>AM2111</td><td>whole-brain imaging</td></tr><tr><td>O.C.T compound</td><td>Sakura Finetek</td><td>4583</td><td></td></tr><tr><td>goat anti-rabbit Alexa Fluro 488</td><td>Invitrogen</td><td>A11008</td><td>Immunohistochemistry</td></tr><tr><td>Cryostat</td><td>Vibratome</td><td>ultrapro 5000</td><td>brain section for IHC</td></tr><tr><td>Evans blue</td><td>Sigma</td><td>E2129</td><td>Detecting vascular perfusion</td></tr><tr><td>Microtome</td><td>Electron Microscopy Sciences</td><td>5000</td><td>brain section for histology</td></tr><tr><td>Avertin (2, 2, 2-Tribromoethanol)</td><td>Sigma</td><td>T48402</td><td>euthanasia</td></tr><tr><td>Fluorescent microscope</td><td>Olympus</td><td>DP73</td><td></td></tr><tr><td>Meloxicam SR</td><td>ZooPharm</td><td></td><td>NSAID analgesia</td></tr></tbody></table>

a thrombotic stroke model based on transient cerebral hypoxia-ischemia

中风的研究已经经历了很多挫折，翻译神经保护疗法进入临床实践。与此相反，在现实世界的治疗剂（tPA溶栓）很少产生机械闭塞基于实验模型的好处，其中占主导地位的临床前中风研究。这板凳和床头之间的分裂意味着需要采用tPA的响应模型在临床前研究中风。为此目的，一个简单的和tPA的反应性血栓性中风模型被发明并在此描述。该模型包括通过在成年小鼠面罩30分钟的单侧颈总动脉和输送7.5％的氧气的瞬态闭塞，同时保持动物的直肠温度在37.5±0.5℃。虽然单侧颈动脉或缺氧的可逆结扎每个抑制脑血流量瞬时只，既侮辱的组合引起再灌注持续赤字，纤维蛋白和血小板沉积，以及大型INFARCT在大脑中动脉提供的领土。重要的是，尾静脉注射重组的tPA在0.5，1，或4小时后THI（10毫克/千克）提供的死亡率和梗死面积的时间依赖性降低。这种新的中风模型简单，并且可以跨实验室标准化比较实验结果。此外，它诱导血栓形成而不骨瓣或引入预形成栓子。鉴于这些独特的优点，在THI模型是一个有益的补充临床前研究中风的剧目。

二维激光散斑对比度成像（LSCI）16，使用由30分钟短暂单侧颈动脉阻塞（tCCAO），30分钟暴露于低氧（7.5％氧气），和30分钟的单方面比较脑血流量（CBF）的改变颈动脉缺氧条件下（THI）结扎。此实验揭示tCCAO在常氧抑制CBF对颈动脉结扎半球〜基线值，迅速 ​​恢复到85％以上的颈动脉阻塞（R 图2A）的释放后的50％。暴露于全身缺氧单独降低了CBF到基线值，返回到常氧气氛（ <str...

Watch this Scientific Journal Video about A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia at JoVE.com

A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia

Thromboembolic stroke models are vital tools for optimizing the recanalization therapy. Here we report a murine thrombotic stroke model based on transient cerebral hypoxic-ischemic (tHI) insult, which triggers thrombosis and infarction, and responds favorably to tissue plasminogen activator (tPA)-mediated fibrinolysis in a therapeutic window similar to those in stroke patients.

基于短暂性脑缺氧缺血性血栓性中风模型

Stroke research has endured many setbacks in translating neuroprotective therapies into clinical practice. In contrast, the real-world therapy (tPA ...

a-thrombotic-stroke-model-based-on-transient-cerebral-hypoxia-ischemia

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

14.7K 次观看.  The Emory University School of Medicine. Thromboembolic stroke models are vital tools for optimizing the recanalization therapy. Here we report a murine thrombotic stroke model based on transient cerebral hypoxic-ischemic (tHI) insult, which triggers thrombosis and infarction, and responds favorably to tissue plasminogen activator (tPA)-mediated fibrinolysis in a therapeutic window similar to those in stroke patients. 

视频: A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia

The Hypoxic Ischemic Encephalopathy Model of Perinatal Ischemia

Hypoxic-Ischemic Encephalopathy (HIE) is the consequence of systemic asphyxia occurring at birth. Twenty five percent of neonates with HIE develop severe and permanent neuropsychological sequelae, including mental retardation, cerebral palsy, and epilepsy. The outcomes of HIE are devastating and permanent, making it critical to identify and develop therapeutic strategies to reduce brain injury in newborns with HIE. To that end, the neonatal rat model for hypoxic-ischemic brain injury has been developed to model this human condition. The HIE model was first validated by Vannucci et al 1 and has since been extensively used to identify mechanisms of brain injury resulting from perinatal hypoxia-ischemia 2 and to test potential therapeutic interventions 3,4. The HIE model is a two step process and involves the ligation of the left common carotid artery followed by exposure to a hypoxic environment. Cerebral blood flow (CBF) in the hemisphere ipsilateral to the ligated carotid artery does not decrease because of the collateral blood flow via the circle of Willis; however with lower oxygen tension, the CBF in the ipsilateral hemisphere decreases significantly and results in unilateral ischemic injury. The use of 2,3,5-triphenyltetrazolium chloride (TTC) to stain and identify ischemic brain tissue was originally developed for adult models of rodent cerebral ischemia 5, and is used to evaluate the extent of cerebral infarctin at early time points up to 72 hours after the ischemic event 6. In this video, we demonstrate the hypoxic-ischemic injury model in postnatal rat brain and the evaluation of the infarct size using TTC staining.

The postnatal rat model for hypoxic-ischemic brain injury is a well-established model of human neonatal hypoxic ischemic encephalopathy (HIE). In this article, we describe the model of HIE in post-natal rat pups.

围产期缺血缺氧缺血性脑病模型

Hypoxic-Ischemic Encephalopathy (HIE) is the consequence of systemic asphyxia occurring at birth. Twenty five percent of neonates with HIE develop severe ...

科研

生物学

Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical Lesion Model for Mouse Stroke Studies

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected light-sensitive dye. Following illumination, the dye is activated and produces singlet oxygen that damages components of endothelial cell membranes, with subsequent platelet aggregation and thrombi formation, which eventually determines the interruption of local blood flow. This approach, initially proposed by Rosenblum and El-Sabban in 1977, was later improved by Watson in 1985 in rat brain and set the basis of the current model. Also, the increased availability of transgenic mouse lines further contributed to raise the interest on the photothrombosis model. Briefly, a photosensitive dye (Rose Bengal) is injected intraperitoneally and enters the blood stream. When illuminated by a cold light source, the dye becomes activated and induces endothelial damage with platelet activation and thrombosis, resulting in local blood flow interruption. The light source can be applied on the intact skull with no need of craniotomy, which allows targeting of any cortical area of interest in a reproducible and non-invasive way. The mouse is then sutured and allowed to wake up. The evaluation of ischemic damage can be quickly accomplished by triphenyl-tetrazolium chloride or cresyl violet staining. This technique produces infarction of small size and well-delimited boundaries, which is highly advantageous for precise cell characterization or functional studies. Furthermore, it is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

Photothrombosis is a quick, minimally-invasive technique for inducing small and well-delimited infarction in areas of interest in highly reproducible manner. It is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

光化学缺血：一种微创和可重复性的光化学皮质病变模型小鼠中风研究

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected ...

医学

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 ...

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 ...

A Fibrin-Enriched and tPA-Sensitive Photothrombotic Stroke Model

An ideal thromboembolic stroke model requires certain properties, including relatively simple surgical procedures with low mortality, a consistent infarction size and location, precipitation of platelet:fibrin intermixed blood clots similar to those in patients, and an adequate sensitivity to fibrinolytic treatment. The rose bengal (RB) dye-based photothrombotic stroke model meets the first two requirements but is highly refractory to tPA-mediated lytic treatment, presumably due to its platelet-rich, but fibrin-poor clot composition. We reason that combination of RB dye (50 mg/kg) and a sub-thrombotic dose of thrombin (80 U/kg) for photoactivation aimed at the proximal branch of middle cerebral artery (MCA) may produce fibrin-enriched and tPA-sensitive clots. Indeed, the thrombin and RB (T+RB)-combined photothrombosis model triggered mixed platelet:fibrin blood clots, as shown by immunostaining and immunoblots, and maintained consistent infarct sizes and locations plus low mortality. Moreover, intravenous injection of tPA (Alteplase, 10 mg/kg) within 2 h post-photoactivation significantly decreased the infarct size in T+RB photothrombosis. Thus, the thrombin-enhanced photothrombotic stroke model may be a useful experimental model to test novel thrombolytic therapies.

Traditional photothrombotic stroke (PTS) models mainly induce dense platelet aggregates of a high resistance to tissue plasminogen activator (tPA)-lytic treatment. Here a modified murine PTS model is introduced by co-injecting thrombin and photosensitive dye for photoactivation. The thrombin-enhanced PTS model produces mixed platelet:fibrin clots and is highly sensitive to tPA-thrombolysis.

纤维蛋白富集和 tPA 敏感的光血栓形成卒中模型

An ideal thromboembolic stroke model requires certain properties, including relatively simple surgical procedures with low mortality, a consistent ...

神经科学

Modeling Stroke in Mice: Focal Cortical Lesions by Photothrombosis

Stroke is a leading cause of death and acquired adult disability in developed countries. Despite extensive investigation for novel therapeutic strategies, there remain limited therapeutic options for stroke patients. Therefore, more research is needed for pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity, and regeneration. Given the inability of in vitro models to reproduce the complexity of the brain, experimental stroke models are essential for the analysis and subsequent evaluation of novel drug targets for these mechanisms. In addition, detailed standardized models for all procedures are urgently needed to overcome the so-called replication crisis. As an effort within the ImmunoStroke research consortium, a standardized photothrombotic mouse model using an intraperitoneal injection of Rose Bengal and the illumination of the intact skull with a 561 nm laser is described. This model allows the performance of stroke in mice with allocation to any cortical region of the brain without invasive surgery; thus, enabling the study of stroke in various areas of the brain. In this video, the surgical methods of stroke induction in the photothrombotic model along with&#160;histological analysis are demonstrated.

Described here is the photothrombotic stroke model, where a stroke is produced through the intact skull by inducing permanent microvascular occlusion using laser illumination after administration of a photosensitive dye.

小鼠模型中风:光血栓形成的焦皮质病变

Stroke is a leading cause of death and acquired adult disability in developed countries. Despite extensive investigation for novel therapeutic strategies, ...

Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options are limited to a small proportion of stroke patients within the first hours after stroke. Novel therapeutic strategies are being extensively investigated, especially to prolong the therapeutic time window. These current investigations include the study of important pathophysiological pathways after stroke, such as post-stroke inflammation, angiogenesis, neuronal plasticity, and regeneration. Over the last decade, there has been increasing concern about the poor reproducibility of experimental results and scientific findings among independent research groups. To overcome the so-called &#8220;replication crisis&#8221;, detailed standardized models for all procedures are urgently needed. As an effort within the &#8220;ImmunoStroke&#8221; research consortium (https://immunostroke.de/), a standardized mouse model of transient middle cerebral artery occlusion (MCAo) is proposed. This model allows the complete restoration of the blood flow upon removal of the filament, simulating the therapeutic or spontaneous clot lysis that occurs in a large proportion of human strokes. The surgical procedure of this &#8220;filament&#8221; stroke model and tools for its functional analysis are demonstrated in the accompanying video.

Different models of middle cerebral artery occlusion (MCAo) are used in experimental stroke research. Here, an experimental stroke model of transient MCAo via the external carotid artery (ECA) is described, which aims to mimic human stroke, in which the cerebrovascular thrombus is removed due to spontaneous clot lysis or therapy.

模拟小鼠中风:通过颈外动脉的短暂性脑中动脉闭塞

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options ...

大脑中动脉闭塞的短暂性脑缺血模型

Transient Middle Cerebral Artery Occlusion Model of Stroke

Stroke stands as a major cause of death or chronic disability globally. Nevertheless, existing optimal treatments are limited to reperfusion therapies during the acute phase of ischemic stroke. To gain insights into stroke physiopathology and develop innovative therapeutic approaches, in vivo rodent models of stroke play a fundamental role. The availability of genetically modified animals has particularly propelled the use of mice as experimental stroke models.
In stroke patients, occlusion of the middle cerebral artery (MCA) is a common occurrence. Consequently, the most prevalent experimental model involves intraluminal occlusion of the MCA, a minimally invasive technique that doesn't require craniectomy. This procedure involves inserting a monofilament through the external carotid artery (ECA) and advancing it through the internal carotid artery (ICA) until it reaches the branching point of the MCA. After a 45 min arterial occlusion, the monofilament is removed to allow reperfusion. Throughout the process, cerebral blood flow is monitored to confirm the reduction during occlusion and subsequent recovery upon reperfusion. Neurological and tissue outcomes are evaluated using behavioral tests and magnetic resonance imaging (MRI) studies.

作者聚焦：通过大脑中动脉闭塞模型评估缺血性卒中损伤 

This protocol describes the model of transient focal cerebral ischemia in mice through intraluminal occlusion of the middle cerebral artery. Additionally, examples of outcome assessment are shown using magnetic resonance imaging and behavioral tests.

脑卒中短暂性中动脉闭塞模型

Stroke stands as a major cause of death or chronic disability globally. Nevertheless, existing optimal treatments are limited to reperfusion therapies ...

基于短暂性脑缺氧缺血性血栓性中风模型

摘要

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围产期缺血缺氧缺血性脑病模型

光化学缺血：一种微创和可重复性的光化学皮质病变模型小鼠中风研究

造型中风小鼠:在远端大脑中动脉永久性混凝

在P10大鼠新生儿中风的短暂大脑中动脉栓塞模型

纤维蛋白富集和 tPA 敏感的光血栓形成卒中模型

小鼠模型中风:光血栓形成的焦皮质病变

模拟小鼠中风:通过颈外动脉的短暂性脑中动脉闭塞

脑卒中短暂性中动脉闭塞模型

脑卒中短暂性中动脉闭塞模型

脑卒中短暂性中动脉闭塞模型