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>手术刀，虹膜镊子手柄＃和11＃刀片，格雷夫斗牛犬钳钳，拉钩，螺丝刀，10μl注射器，26号斜口针（世界精密仪器公司，萨拉索塔，佛罗里达州，美国） </li><li>微电机的演习和立体的持有人，精髓立体定向喷油器（Stoelting，伍德戴尔，IL，USA） </li><li> 1.0毫米的圆形钻钻，1.0毫米的倒锥钻牙钻（Roboz手术仪器有限公司，公司，盖瑟斯堡，MD，USA） </li></ol></li><li>手术用品<ol class="lalpha"><li>安装螺丝0-80 X 3/32，2.4毫米轴的长度，21号导管4mm长的基座下方，套管假的（塑料，罗阿诺克，VA，USA） </li><li>李连杰义齿丙烯酸树脂和液体的（郎牙科制造有限公司，公司，威灵，IL，USA） </li><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>电推剪（奥斯特，普罗维登斯，RI，USA） </li></ol></li><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><li>盐酸丁丙诺啡（赫士睿公司的Lake Forest，IL，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>光纤照明器（TechniQuip公司，利弗莫尔，CA，USA） </li></ol></li><li>激光多普勒血流仪系统（的ADInstruments公司，科罗拉多斯普林斯，CO，USA） <ol class="lalpha"><li>圣andard铅笔探头</li><li>探头支架</li><li>血液流量计</li><li> PowerLab系统4/30与LabChart 7 </li></ol></li><li>梗死体积的测量<ol class="lalpha"><li>大鼠大脑基质（Zivic - 米勒实验室，公司，艾里逊公园，PA，USA） </li><li> 2,3,5 - 三苯四唑氯化物（Sigma-Aldrich公司，圣路易斯，密苏里州，美国），在PBS中稀释至0.05％ </li><li>平板式扫描仪（爱普生Perfection V30，爱普生美国公司，长滩，CA，USA） </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>麻醉诱导与4％异氟烷在100％O 2的混合气体的感应室中，直到不再大鼠提款后部爪夹送。 </li><li>一化粪池在此过程中，应保持技术，包括使用无菌手套，无菌外科手术器械，一个无菌手术铺巾11。 </li><li>电推剪剃光头顶。 </li><li>大鼠置于俯卧趴在一个温度控制的操作表面（热毯）的吸收垫的位置上。 </li><li>头被放置在立体定向装置放置气体麻醉面罩开始。 </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>润滑油眼药膏的双眼，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>头牢牢的立体的移动设备，手术区洁净交替2％洗必泰和盐水洗涤三次。 </li></ol> 2。手术步骤<ol><li>中线切口用手术刀，是由覆盖在颅骨的眼睛从最尾端的方面（鼻根）之间的耳（上项线）在皮肤上。 </li><li> 3斗牛犬夹横向的皮肤，然后缩回。 </li><li>从颅骨，使多个结构可以清楚地看出，用干棉签去除结缔组织。这些措施包括，冠状缝，前囟门和右侧头骨脊。使用棉签清除血液从手术领域。 </li><li>使用手术显微镜，前囟门的位置，立体机械手进行调整，直到锁定在1.0毫米的圆形钻钻上的前囟门。 </li><li>钻牙钻，然后移动到1.6毫米前侧面，前囟门和5.2毫米。 </li><li>车针，穿透颅骨上钻洞卡恩乌拉的位置（ 图1）。不断地用棉签清除过量的碎片和血液。 </li></ol>在这一点上，一个导向套管可以被插入（步骤7）或ET-1直接注射通过牙钻通孔，可以进行（直接前进到步骤16）。 <ol start="7"><li>接着，钻的孔的3个安装螺丝钻颅骨使用1.0毫米的倒锥钻牙钻（ 图1）通过部分的厚度。在每个约1-2毫米的冠状缝和矢状缝1-2毫米的横向前额骨上钻一个孔。在顶骨钻一个孔，约2-3毫米的冠状缝和2-3毫米，侧的矢状缝同侧导管牙钻洞的后方。三0-80×3/32安装螺丝被放置在这些牙钻洞，并在套管水泥控股将提供支持。螺丝应仅是先进的2或3圈，以便不损坏硬膜。 </lI&gt; <li>导管被放置在立体定位的套管夹持器，并位于前囟。 </li><li>立体机械手调整，直到锁定在导管前囟门。套管移动到牙钻孔位于1.6毫米前壁和5.2毫米的横向于前囟。 </li><li>最后，在导向套管被降低到牙钻孔4.5毫米腹侧囟与最后的前端位置。 </li><li>激光多普勒血流探测位置（可选） <ol class="lalpha"><li>为了使用LDF监测大脑的血流量，探头支架可以放置到位置加盖指南套管用牙科水泥。 </li><li>除了小楔形"选项卡，探头支架基座的底座齐平修剪。 </li><li>探针架座，然后放置后导向套管和公正的内侧与面向的标签的内侧（ 图1）的侧颅底脊。 </li><li>探针架座和g指南套管贴在一起，用牙科水泥。 </li></ol></li><li>然后，使用牙科用粘固剂的插管固定到位。水泥是在三个螺钉与所有的接触和围绕所述套管的整个基地。 </li><li>应完全干燥的水泥之前除去的套管夹持器。这大约需要5分钟。 </li></ol>在这些步骤之后，手术切口可以关闭，可拧入导向套管和套管虚设。另外，ET-1诱导MCAO可以对大鼠进行一段时间后，从插管植入手术的恢复。对于此方法，应接着执行步骤19，并在稍后的时间，可以执行步骤14-18。要执行同样的手术过程中引导导管植入和注射ET-1，第14步进行。 <ol start="14"><li>输液注射器装入与ET-1（在PBS中稀释至80μM），然后安装在立体定向喷射器。 </li><li> "立体定位的操纵器被调整直到针尖在导向套管的边缘被归零。 </li><li>针尖降低，通过导向插管17.2毫米腹侧的导向套管的边缘的位置的。如果不使用的导向套管，针尖归零在囟和降低通过牙钻孔的位置8.7毫米腹侧囟。 </li><li> 3微升80μMET1注入的速率（每分钟）的1微升。 </li><li>注射器为3分钟，在输注后是完整的，然后慢慢地去除留在原地。 </li><li>切口6.0尼龙缝线被拧入插管和插管虚设封闭。 </li><li> （ 即丁丙诺啡在0.05-0.1毫克/公斤），应使用适当的镇痛药的剂量，手术后的恢复期间，以减少疼痛和不适。 </li><li>大鼠从手术室被移除，并放置在温暖，干燥的恢复区，以防止低温，柔软的食物和水的自由，方便地访问。 </李&gt; </ol>

<ol>
	<li>Stroke--1989. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recommendations+on+stroke+prevention,+diagnosis,+and+therapy.+Report+of+the+WHO+Task+Force+on+Stroke+and+other+Cerebrovascular+Disorders.">Recommendations on stroke prevention, diagnosis, and therapy. Report of the WHO Task Force on Stroke and other Cerebrovascular Disorders.</a> Stroke. 20, 1407-1431 (1989).</li><li>Lloyd-Jones, D., 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=Heart+disease+and+stroke+statistics--2009+update:+a+report+from+the+American+Heart+Association+Statistics+Committee+and+Stroke+Statistics+Subcommittee.">Heart disease and stroke statistics--2009 update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee.</a> Circulation. 119, 480-486 (2009).</li><li>Mohr, J. P., Gautier, J. C., Hier, D., Stein, R. W., Barnett, H. J. M., Stein, B. M., Mohr, J. P., Yatsu, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Stroke: pathophysiology, diagnosis, and management. , 377-450 (1986).</li><li>Robinson, M. J., Macrae, I. M., Todd, M., Reid, J. L., McCulloch, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+local+cerebral+blood+flow+to+pathological+levels+by+endothelin-1+applied+to+the+middle+cerebral+artery+in+the+rat.">Reduction of local cerebral blood flow to pathological levels by endothelin-1 applied to the middle cerebral artery in the rat.</a> Neurosci. Lett. 118, 269-272 (1990).</li><li>Sharkey, J., Ritchie, I. M., Kelly, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perivascular+microapplication+of+endothelin-1:+a+new+model+of+focal+cerebral+ischaemia+in+the+rat.">Perivascular microapplication of endothelin-1: a new model of focal cerebral ischaemia in the rat.</a> J. Cereb. Blood Flow Metab. 13, 865-871 (1993).</li><li>O'Neill, M. J., Clemens, J. A. <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+cerebral+ischemia.">Rodent models of focal cerebral ischemia.</a> Curr. Protoc. Neurosci. Chapter 9 (Unit 9), (2001).</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>Braeuninger, S., Kleinschnitz, C. <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+cerebral+ischemia:+procedural+pitfalls+and+translational+problems.">Rodent models of focal cerebral ischemia: procedural pitfalls and translational problems.</a> Exp. Transl. Stroke Med. 1, 8 (2009).</li><li>Tomsick, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intravenous+thrombolysis+for+acute+ischemic+stroke.">Intravenous thrombolysis for acute ischemic stroke.</a> J. Vasc. Interv. Radiol. 15, 67-76 (2004).</li><li>Olsen, T. S., Lassen, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+dynamic+concept+of+middle+cerebral+artery+occlusion+and+cerebral+infarction+in+the+acute+state+based+on+interpreting+severe+hyperemia+as+a+sign+of+embolic+migration.">A dynamic concept of middle cerebral artery occlusion and cerebral infarction in the acute state based on interpreting severe hyperemia as a sign of embolic migration.</a> Stroke. 15, 458-468 (1984).</li><li>Pritchett-Corning, K. R., Luo, Y., Mulder, G. B., White, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+of+rodent+surgery+for+the+new+surgeon.">Principles of rodent surgery for the new surgeon.</a> J. Vis. Exp. (47), e2586 (2011).</li><li>Gonzalez, C. L., Kolb, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+different+models+of+stroke+on+behaviour+and+brain+morphology.">A comparison of different models of stroke on behaviour and brain morphology.</a> Eur. J. Neurosci. 18, 1950-1962 (2003).</li><li>Ansari, S., Azari, H., McConnell, D. J., Afzal, A., Mocco, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraluminal+middle+cerebral+artery+occlusion+(MCAO)+model+for+ischemic+stroke+with+laser+doppler+flowmetry+guidance+in+mice.">Intraluminal middle cerebral artery occlusion (MCAO) model for ischemic stroke with laser doppler flowmetry guidance in mice.</a> J. Vis. Exp. (51), e2879 (2011).</li><li>Sharkey, J., Butcher, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterisation+of+an+experimental+model+of+stroke+produced+by+intracerebral+microinjection+of+endothelin-1+adjacent+to+the+rat+middle+cerebral+artery.">Characterisation of an experimental model of stroke produced by intracerebral microinjection of endothelin-1 adjacent to the rat middle cerebral artery.</a> J. Neurosci. Methods. 60, 125-131 (1995).</li><li>Macrae, I. M., Robinson, M. J., Graham, D. I., Reid, J. L., McCulloch, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelin-1-induced+reductions+in+cerebral+blood+flow:+dose+dependency,+time+course,+and+neuropathological+consequences.">Endothelin-1-induced reductions in cerebral blood flow: dose dependency, time course, and neuropathological consequences.</a> J. Cereb. Blood Flow Metab. 13, 276-284 (1993).</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 (1-7), 1084-1096 (2011).</li><li>Fisher, M., 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=Update+of+the+stroke+therapy+academic+industry+roundtable+preclinical+recommendations.">Update of the stroke therapy academic industry roundtable preclinical recommendations.</a> Stroke. 40, 2244-2250 (2009).</li></ol>

没有利益冲突的声明。

ET-1诱导脑缺血是一个既定的模式，经常使用在多个品系的实验缺血性中风。许多变量，如大鼠株，动物年龄，体温，麻醉方法和操作者的专业知识，可导致梗死体积的变异性增加时，使用此模型5中，14。然而，一些研究者已经证明，这种模式的优势，包括相对非侵入性的方法，剂量反应的脑血流ET-1，和能力，以避免完全由麻醉注射ET-1在清醒大鼠14日，15。值得注意的是，有很多...

<table border="1"><tbody><tr><td></td><td></td><td></td><td><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>手术刀，虹膜镊子手柄＃和11＃刀片，格雷夫斗牛犬钳钳，拉钩，螺丝刀，10μl注射器，26号斜口针（世界精密仪器公司，萨拉索塔，佛罗里达州，美国） </li><li>微电机的演习和立体的持有人，精髓立体定向喷油器（Stoelting，伍德戴尔，IL，USA） </li><li> 1.0毫米的圆形钻钻，1.0毫米的倒锥钻牙钻（Roboz手术仪器有限公司，公司，盖瑟斯堡，MD，USA） </li></ol></li><li>手术用品<ol class="lalpha"><li>安装螺丝0-80 X 3/32，2.4毫米轴的长度，21号导管4mm长的基座下方，套管假的（塑料件，罗阿诺克，VA，USA） </li><li>李连杰义齿丙烯酸树脂和液体的（郎牙科制造有限公司，公司，威灵，IL，USA） </li><li> 3.0尼龙线（绿洲，Mettawa，IL，USA） </li><li>棉签，Puralube的眼膏（费舍尔科学性C，匹兹堡，PA，USA） </li><li>电推剪（奥斯特，普罗维登斯，RI，USA） </li></ol></li><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><li>盐酸丁丙诺啡（赫士睿公司的Lake Forest，IL，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>光纤照明器（TechniQuip公司，利弗莫尔，CA，USA） </li></ol></li><li>激光多普勒血流仪系统（的ADInstruments公司，科罗拉多斯普林斯，CO，USA） <ol class="lalpha"><li>标准铅笔探头</li><li>探头支架</li><li>血液流量计</li><li> PowerLab系统4/30与LabChart 7 </li></ol></li><li>梗死体积的测量<ol class="lalpha"><li>大鼠脑矩阵（Zivic - 米勒实验室，公司，艾里逊公园，PA，USA） </li><li> 2,3,5 - 三苯四唑氯化物（Sigma-Aldrich公司，圣路易斯，密苏里州，美国），在PBS中稀释至0.05％ </li><li>平板式扫描仪（爱普生Perfection V30，爱普生美国公司，长滩，CA，USA） </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></td></tr></tbody></table>

endothelin-1 induced middle cerebral artery occlusion model for ischemic stroke with laser doppler flowmetry guidance in rat

2009年1，2，耗资约70亿美元在美国，中风是导致残疾和死亡的第三大原因，在世界上的头号。已经开发了几种型号的脑缺血，中风模仿人类的​​条件。曾有人建议，高达80％的所有笔划的结果由缺血性脑损伤中的大脑中动脉（MCA）的第3区。在20世纪90年代早期，内皮素-1（ET-1）4，通过将它直接相邻的表面后，在MCA开颅用于诱导缺血。后来，这个模型进行了改进，采用立体定向注射ET-1的相邻MCA局灶性脑缺血。这种模式的主要优点包括能够迅速执行过程，控制动脉收缩，通过改变剂量的ET-1交付的能力，没有需要处理的颅外血管供应血液输送到大脑，并逐渐reperfusi的利率，更接近人类再灌注损伤的5-7。另一方面，ET-1的模型具有的缺点，其中包括需要开颅，以及更高的变异性的每搏输出量8。这种变化可以减少使用激光多普勒血流仪（LDF），以验证脑缺血时ET-1次静脉滴注。变异性中风的因素影响包括输液和一批ET-1的精度，使用了6个 。另一个重要因素是，虽然再灌注损伤是一种常见的发生在人类中风，阻断ET-1诱导脑缺血的持续时间可能不会紧密地模仿人类中风病人多有局部灌注在一个小时内闭塞9天， 10。此协议中详细描述的ET-1诱导的脑缺血模型大鼠缺血性中风的。在整个过程中，它也将提请注意的特殊考虑和潜在的缺点。

1。术后神经学评估动物苏醒后，可用于广泛的测试，以评估神经功能障碍，包括平衡，握力​​，爪子配售，姿势不对称和楼梯攀登。葵花籽任务的运动和感觉功能，具有显着的相关性梗死第7卷，12是一个总的评价。在这个任务中，老鼠是定时开放和消费5葵花籽。五个种子放置在一个角落里，一个空的，干燥，塑料保持架和操纵的种子所花费的时间被记录。?...

Watch this Scientific Journal Video about Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat at JoVE.com

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

已经开发了几种脑缺血的动物模型来模拟人的条件中风。该协议描述了内皮素-1（ET-1）诱导大鼠大脑中动脉闭塞（MCAO）模型大鼠缺血性中风。此外，重要的考虑因素，此模型的优点和缺点，进行了讨论。

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 ...

endothelin-1-induced-middle-cerebral-artery-occlusion-model-for

Research

JoVE Journal

Medicine

19.9K 次观看.  University of Florida. 已经开发了几种脑缺血的动物模型来模拟人的条件中风。该协议描述了内皮素-1（ET-1）诱导大鼠大脑中动脉闭塞（MCAO）模型大鼠缺血性中风。此外，重要的考虑因素，此模型的优点和缺点，进行了讨论。

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

Focal Cerebral Ischemia Model by Endovascular Suture Occlusion of the Middle Cerebral Artery in the Rat

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable clinical state; in the development of animal models of focal ischemia, however, achieving reproducibility of experimentally induced infarct volume is essential. The rat is a widely used animal model for stroke due to its relatively low animal husbandry costs and to the similarity of its cranial circulation to that of humans2,3. In humans, the middle cerebral artery (MCA) is most commonly affected in stroke syndromes and multiple methods of MCA occlusion (MCAO) have been described to mimic this clinical syndrome in animal models. Because recanalization commonly occurs following an acute stroke in the human, reperfusion after a period of occlusion has been included in many of these models. In this video, we demonstrate the transient endovascular suture MCAO model in the spontaneously hypertensive rat (SHR). A filament with a silicon tip coating is placed intraluminally at the MCA origin for 60 minutes, followed by reperfusion. Note that the optimal occlusion period may vary in other rat strains, such as Wistar or Sprague-Dawley. Several behavioral indicators of stroke in the rat are shown. Focal ischemia is confirmed using T2-weighted magnetic resonance images and by staining brain sections with 2,3,5-triphenyltetrazolium chloride (TTC) 24 hours after MCAO.

Surgical induction of ischemic brain damage in the rat is a widely used model for stroke research. Here we demonstrate the induction of focal cerebral ischemia by occlusion of the middle cerebral artery. Visualization of the resulting infarct by histological staining and magnetic resonance imaging is also shown.

焦距脑缺血模型大鼠大脑中动脉的血管内缝合闭塞

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable ...

科研

神经科学

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 ...

医学

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 ...

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

Ischemic stroke leads to vasogenic cerebral edema and subsequent primary brain injury, which is mediated through destruction of the blood-brain barrier (BBB). Rats with induced ischemic stroke were established and used as in vivo models to investigate the functional integrity of the BBB. Spectrophotometric detection of Evans blue (EB) in the brain samples with ischemic injury could provide reliable justification for the research and development of novel therapeutic modalities. This method generates reproducible results, and is applicable in any laboratory without a need for special equipment. Here, we present a visualized and technical guideline on the detection of the extravasation of EB following induction of ischemic stroke in rats.

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

The overall goal of this procedure is to provide a highly reproducible technique for in vivo assessment of the blood-brain barrier disruption in rat models of ischemic stroke.

一种体内评价大鼠缺血性中风模型中血脑屏障的破坏

Ischemic stroke leads to vasogenic cerebral edema and subsequent primary brain injury, which is mediated through destruction of the blood-brain barrier ...

Laser-Induced Brain Injury in the Motor Cortex of Rats

A common technique for inducing stroke in experimental rodent models involves the transient (often denoted as MCAO-t) or permanent (designated as MCAO-p) occlusion of the middle cerebral artery (MCA) using a catheter. This generally accepted technique, however, has some limitations, thereby limiting its extensive use. Stroke induction by this method is often characterized by high variability in the localization and size of the ischemic area, periodical occurrences of hemorrhage, and high death rates. Also, the successful completion of any of the transient or permanent procedures requires expertise and often lasts for about 30 minutes. In this protocol, a laser irradiation technique is presented that can serve as an alternative method for inducing and studying brain injury in rodent models.
When compared to rats in the control and MCAO groups, the brain injury by laser induction showed reduced variability in body temperature, infarct volume, brain edema, intracranial hemorrhage, and mortality. Furthermore, the use of a laser-induced injury caused damage to the brain tissues only in the motor cortex unlike in the MCAO experiments where destruction of both the motor cortex and striatal tissues is observed.
Findings from this investigation suggest that laser irradiation could serve as an alternative and effective technique for inducing brain injury in the motor cortex. The method also shortens the time for completing the procedure and does not require expert handlers.

The protocol presented here shows a technique to create a rodent model of brain injury. The method described here uses laser irradiation and targets motor cortex.

大鼠运动皮层中的激光诱发脑损伤

A common technique for inducing stroke in experimental rodent models involves the transient (often denoted as MCAO-t) or permanent (designated as MCAO-p) ...

A Murine Model of Ischemic Retinal Injury Induced by Transient Bilateral Common Carotid Artery Occlusion

Diverse vascular diseases such as diabetic retinopathy, occlusion of retinal veins or arteries and ocular ischemic syndrome can lead to retinal ischemia. To investigate pathological mechanisms of retinal ischemia, relevant experimental models need to be developed. Anatomically, a main retinal blood supplying vessel is the ophthalmic artery (OpA) and OpA originates from the internal carotid artery of the common carotid artery (CCA). Thus, disruption of CCA could effectively cause retinal ischemia. Here, we established a mouse model of retinal ischemia by transient bilateral common carotid artery occlusion (tBCCAO) to tie the right CCA with 6-0 silk sutures and to occlude the left CCA transiently for 2 seconds via a clamp, and showed that tBCCAO could induce acute retinal ischemia leading to retinal dysfunction. The current method reduces reliance on surgical instruments by only using surgical needles and a clamp, shortens occlusion time to minimize unexpected animal death, which is often seen in mouse models of middle cerebral artery occlusion, and maintains reproducibility of common retinal ischemic findings. The model can be utilized to investigate the pathophysiology of ischemic retinopathies in mice and further can be used for in vivo drug screening.

Here, we describe a mouse model of retinal ischemia by transient bilateral common carotid artery occlusion using simple sutures and a clamp. This model can be useful for understanding the pathological mechanisms of retinal ischemia caused by cardiovascular abnormalities.

由瞬态双边常见胡萝卜动脉闭塞引起的缺血性视网膜损伤的 Murine 模型

Diverse vascular diseases such as diabetic retinopathy, occlusion of retinal veins or arteries and ocular ischemic syndrome can lead to retinal ischemia. ...

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 ...

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

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此视频中的章节

焦距脑缺血模型大鼠大脑中动脉的血管内缝合闭塞

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

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

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

一种体内评价大鼠缺血性中风模型中血脑屏障的破坏

大鼠运动皮层中的激光诱发脑损伤

由瞬态双边常见胡萝卜动脉闭塞引起的缺血性视网膜损伤的 Murine 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

脑卒中短暂性中动脉闭塞模型