Name: photothrombotic ischemia: a minimally invasive and reproducible photochemical cortical lesion model for mouse stroke studies
Uploaded: 2013-06-09T14:00:00
Description: Watch this Scientific Journal Video about Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical Lesion Model for Mouse Stroke Studies at JoVE.com

我们感谢安娜丽莎滑稽的，有见地的建议和意见，Maurizio Grassano的滨海Boido和埃尔米拉Pajaj拍摄。这项工作是由FP7-MC-214003-2（居里夫人首次培训网络AXREGEN）和COMPAGNIA迪圣保罗，gliarep项目。

1。手术前<ol><li>称量玫瑰红的1.5毫升管中，溶解在无菌生理盐水溶液中，直到达到最终浓度为15毫克/毫升。通过0.2微米的过滤器过滤灭菌，并将其存储在黑暗中，在室温下长达两个月。 </li><li>所有的手术器械消毒灭菌。应消毒手术区不到一小时才开始手术。 </li><li>记录小鼠体重调整剂量注入孟加拉玫瑰红。女CD1小鼠12周龄本协议注入10微升/克动物体重。孟加拉玫瑰红需要的量，以产生所需的皮...

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	<li>Lopez, A. D., Mathers, C. D., Ezzati, M., Jamison, D. T., Murray, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+and+regional+burden+of+disease+and+risk+factors.">Global and regional burden of disease and risk factors.</a> Lancet. 367, 1747-1757 (2001).</li><li>Mathers, C. D., Boerma, T., Ma Fat, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+and+regional+causes+of+death.">Global and regional causes of death.</a> Br. Med. Bull. 92, 7-32 (2009).</li><li>Rosenblum, W. 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Neurol. 17, 497-504 (1985).</li><li>Bergeron, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inducing+photochemical+cortical+lesions+in+rat+brain.">Inducing photochemical cortical lesions in rat brain.</a> Curr. Protoc. Neurosci. Chapter 9, Unit 9 16 (2003).</li><li>Lee, J. K., 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=Photochemically+induced+cerebral+ischemia+in+a+mouse+model.">Photochemically induced cerebral ischemia in a mouse model.</a> Surg. Neurol. 67, 620-625 (2007).</li><li>Dietrich, W. D., Watson, B. D., Busto, R., Ginsberg, M. D., Bethea, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photochemically+induced+cerebral+infarction.+I.+Early+microvascular+alterations.">Photochemically induced cerebral infarction. I. Early microvascular alterations.</a> Acta Neuropathol. 72, 315-325 (1987).</li><li>Schroeter, M., Jander, S., Stoll, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+induction+of+focal+cerebral+ischemia+in+mice+by+photothrombosis+of+cortical+microvessels:+characterization+of+inflammatory+responses.">Non-invasive induction of focal cerebral ischemia in mice by photothrombosis of cortical microvessels: characterization of inflammatory responses.</a> J. Neurosci. 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Korean Neurosurg. Soc. 46, 479-483 (2009).</li><li>Silva, V. M., Corson, N., Elder, A., Oberdorster, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+rat+ear+vein+model+for+investigating+in+vivo+thrombogenicity+of+ultrafine+particles+(UFP).">The rat ear vein model for investigating in vivo thrombogenicity of ultrafine particles (UFP).</a> Toxicol. Sci. 85, 983-989 (2005).</li><li>Watson, B. D., Prado, R., Dietrich, W. D., Ginsberg, M. D., Green, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photochemically+induced+spinal+cord+injury+in+the+rat.">Photochemically induced spinal cord injury in the rat.</a> Brain Res. 367, 296-300 (1986).</li><li>Van Reempts, J., Van Deuren, B., Van de Ven, M., Cornelissen, F., Borgers, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flunarizine+reduces+cerebral+infarct+size+after+photochemically+induced+thrombosis+in+spontaneously+hypertensive+rats.">Flunarizine reduces cerebral infarct size after photochemically induced thrombosis in spontaneously hypertensive rats.</a> Stroke. 18, 1113-1119 (1987).</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>Kleinschnitz, C., 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=Blocking+of+platelets+or+intrinsic+coagulation+pathway-driven+thrombosis+does+not+prevent+cerebral+infarctions+induced+by+photothrombosis.">Blocking of platelets or intrinsic coagulation pathway-driven thrombosis does not prevent cerebral infarctions induced by photothrombosis.</a> Stroke. 39, 1262-1268 (2008).</li><li>Porritt, M. J., 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=Photothrombosis-induced+infarction+of+the+mouse+cerebral+cortex+is+not+affected+by+the+Nrf2-activator+sulforaphane.">Photothrombosis-induced infarction of the mouse cerebral cortex is not affected by the Nrf2-activator sulforaphane.</a> PLoS One. 7, e41090 (2012).</li><li>Baskin, Y. K., Dietrich, W. D., Green, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+effective+behavioral+tasks+for+evaluating+sensorimotor+dysfunction+following+traumatic+brain+injury+in+mice.">Two effective behavioral tasks for evaluating sensorimotor dysfunction following traumatic brain injury in mice.</a> J. Neurosci Methods. 129, 87-93 (2003).</li><li>Markgraf, C. G., 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=Comparative+histopathologic+consequences+of+photothrombotic+occlusion+of+the+distal+middle+cerebral+artery+in+Sprague-Dawley+and+Wistar+rats.">Comparative histopathologic consequences of photothrombotic occlusion of the distal middle cerebral artery in Sprague-Dawley and Wistar rats.</a> Stroke. 24, 286-292 (1993).</li><li>Wester, P., Watson, B. D., Prado, R., Dietrich, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+photothrombotic+'ring'+model+of+rat+stroke-in-evolution+displaying+putative+penumbral+inversion.">A photothrombotic 'ring' model of rat stroke-in-evolution displaying putative penumbral inversion.</a> Stroke. 26, 444-450 (1995).</li><li>Hu, X., Wester, P., Brannstrom, T., Watson, B. D., Gu, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progressive+and+reproducible+focal+cortical+ischemia+with+or+without+late+spontaneous+reperfusion+generated+by+a+ring-shaped,+laser-driven+photothrombotic+lesion+in+rats.">Progressive and reproducible focal cortical ischemia with or without late spontaneous reperfusion generated by a ring-shaped, laser-driven photothrombotic lesion in rats.</a> Brain Res. Brain Res. Protoc. 7, 76-85 (2001).</li><li>Maxwell, K. A., Dyck, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+reproducible+focal+ischemic+lesions+in+neonatal+mice+by+photothrombosis.">Induction of reproducible focal ischemic lesions in neonatal mice by photothrombosis.</a> Dev. Neurosci. 27, 121-126 (2005).</li><li>Kuroiwa, T., 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=Development+of+a+rat+model+of+photothrombotic+ischemia+and+infarction+within+the+caudoputamen.">Development of a rat model of photothrombotic ischemia and infarction within the caudoputamen.</a> Stroke. 40, 248-253 (2009).</li></ol>

修改和替换由于其在562 nm处的吸收峰从过滤的氙弧灯，绿光激光器最初被选择照射在感光玫瑰红。虽然仍然使用激光介导激发recently5，它可以取代冷光源灯泡，也能确保染料激发10,15。冷光源光纤更易于操作，更便宜比激光源。然而，应该注意的是激光器被普遍用来针对单个表面小动脉开颅手术后在体内的特定血管凝血10。 交付敏感?...

在21世纪的开始，缺血性中风是一种破坏性疾病，长期残疾和全球死亡原因的第二位，其中中风占约5,7万人死亡，在2004年2代表第二个原因。尽管被放在许多努力，但仍然没有有效的治疗方法，可改善卒中后功能恢复。卒中动物模型被广泛用于中风研究的领域中，因为它们允许缺血性损伤的病理生理学机制的建模和测试不同的神经保护策略在体内的疗效。这些模型大多通过中断（暂时或永久）大脑中动脉内的血流量，而其他车型的开发，研究在特定区域，通常是电动机和体感皮层的病变体积小，旨在引起广泛梗死。然而，有几个因素可能会导致产生交流在实验中风的研究，其中包括所使用的小鼠品系ERTAIN变异程度，在研究中的动物的年龄和性别，并最重要的是，该技术通过诱导局部缺血损伤。对于后一点，手术的持续时间和侵袭性的（ 即开颅手术的需要），以及需要操作者能够可靠地诱导缺血性病变的外科手术技术人员的关键决定因素，一个成功的体内中风研究的相关。 最初提出的概念photothrombosis布鲁姆和El-萨班在1977年3和其应用在大鼠脑沃森等人成为著名的。于1985年4技术大幅度提高，并设置当前模型3的基础- 6。光化学方法的目的是通过光激活的光敏感染料先前交付进入血液系统，WH诱发皮质梗死ICH导致局部血管血栓形成暴露在光天化日之下的地区。当循环染料在适当的波长的照明冷光源，它释放出能量的氧分子，从而能够产生大量的反应性高的单重态氧产品。这些氧中间体诱导血管内皮细胞的膜脂质过氧化，导致血小板粘附和聚集，并最终确定局部脑血流量中断7的血栓的形成。 photothrombosis是一个非规范性脑缺血模型，不吸留或打破只有一个动脉因为它通常发生在人类卒中，但诱导病变的比较浅的容器，这会导致在暴露在光线下的区域中选择性的血流中断。由于这个原因，这种方法可能适合皮层可塑性的细胞和分子的研究。这种技术的主要优点驻留在其简单的执行。此外，photothrombosis可以容易地进行在每只动物约40分钟，包括21分钟的等待时间（3分钟麻醉; 1分钟到剃的头皮;动物放置在立体定位仪上3至5分钟，2分钟，擦洗头皮消毒液，做一个切口，清洁的头骨; 2到4分钟，将冷光源光纤，1分钟，注入孟加拉玫瑰红溶液，5分钟等待腹腔扩散，15分钟的照明; 5分钟，清洗伤口，缝合动物）。此外，没有手术专业知识是必要执行此技术通过简单的照明完整的头骨，引起病变。不同于古典动脉闭塞，这个方法决定选择性地照射区域内的软脑膜和脑实质微血管闭塞，没有侧支血管留在目标区域的氧气供应，降低病变之间的变异。 其特殊的性质，尽管光化学损伤股脑中风发生的重要机制。类似于人类卒中的动脉闭塞，在照射区域7的血小板聚集和凝块的形成决定的血流中断。同样，这种模式也分享必不可少的炎症反应，如在大脑中动脉闭塞8。然而，因为以及delimitated的边界，半影区，它对应于一个区域的部分保存下来的代谢，是非常减少或不存在的后光化学病变。这种边界清晰，可以方便的在缺血性或完整的皮质区细胞反应研究。 photothrombosis小鼠模型是特别适合于在各种转基因动物的中风研究。事实上，经典机型不适合所有菌株和长时间的研究在C57BL / 6小鼠品系死亡率高的比例，可能会导致偏见9。

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 <td>MATERIAL NAME</td>
 <td>COMPANY </td>
 <td>CATALOGUE NUMBER</td>
 <td></td>
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 <td></td>
 <td></td>
 <td></td>
 <td>Solutions and chemicals</td>
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 <td>Rose Bengal </td>
 <td>Sigma, Italy </td>
 <td>330000 </td>
 <td></td>
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 <td>Isoflurane Vet </td>
 <td>Merial </td>
 <td> 103120022 </td>
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 <td>Betadine </td>
 <td>Asta Medica </td>
 <td></td>
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 <td>Paraformaldehyde </td>
 <td>Sigma-Aldrich </td>
 <td> 158127 </td>
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 <td></td>
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 <td>Surgical material and equipment</td>
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 <td>Fluosorber Filter </td>
 <td>Havard apparatus </td>
 <td> 340415 </td>
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 <td>150W fiber optic illuminator</td>
 <td>Photonic</td>
 <td>PL3000</td>
 <td></td>
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 <td>Temperature Controller for Plate TCAT-2DF </td>
 <td>Havard apparatus</td>
 <td>727561 </td>
 <td></td>
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 <td>Stereotaxic Instrument </td>
 <td>Stoelting</td>
 <td> 51950 </td>
 <td></td>
 </tr>
 <tr>
 <td>Operating microscope </td>
 <td>Takagi </td>
 <td> OM8 </td>
 <td></td>
 </tr>
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 <td>Heating pad </td>
 <td></td>
 <td></td>
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 <td>Oxygen and nitrogen gas</td>
 <td></td>
 <td></td>
 <td></td>
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 <td>Surgery Tools</td>
 <td>World precision instrument </td>
 <td></td>
 <td>Optic fiber taps and mask are custom-made </td>
 </tr>
 </tbody>
</table>

photothrombotic ischemia: a minimally invasive and reproducible photochemical cortical lesion model for mouse stroke studies

光化学中风模型的目的是通过在一个给定的皮层区的先前注入的光敏感的染料的光激活诱导的局部缺血损伤。照明，染料被激活，并产生单重态氧，损害元件的内皮细胞的膜，随后的血小板聚集和血栓形成，最终确定了局部血流量的中断。布鲁姆和El-萨班于1977年，最初提出这种方法，后来在1985年提高沃森在大鼠脑并设置当前模型的基础。此外，供应量增加的转基因小鼠线进一步提供的提高利息上photothrombosis模型。简单地说，光敏染料（玫瑰红）腹膜内注射，并进入血流。当冷光源照明，染料被激活，诱导血管内皮损伤，血小板活化和血栓形成，造成局部血流中断。光源可应用于完整的头骨，无需开颅手术，可以针对任何皮质区的利益在一个可重复的和非侵入性的方式。鼠标，然后缝合，并允许醒来。局部缺血损伤的评价可以迅速通过三苯基四唑鎓氯化物或甲酚紫染色。该技术生产出体积小，以及分隔的边界，这是非常有利于精确的电池特性或功能的研究梗死。此外，它是特别适合用于研究细胞和分子在转基因小鼠中脑的可塑性响应相关。

此协议将产生皮质病变，皮质解剖后，已是可见的，肉眼（ 图1A-1C）。光化学病变发展浅表和深部皮质层组织是半透明到足以允许孟加拉玫瑰红的光激活的。脑梗塞的程度的测量，可快速地执行通过组织染色与三苯基氯化四氮唑（TTC）在4％多聚甲醛（PFA）固定后对新鲜组织或甲酚紫。 TTC孵育之前，新鲜解剖的脑定位在脑切片机，划分成1毫米厚的片（ 图1C）。 TTC标记为?...

Watch this Scientific Journal Video about Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical Lesion Model for Mouse Stroke Studies at JoVE.com

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

的Photothrombosis是一种快速，微创技术领域的兴趣，高度可重复的方式诱导小以及分隔梗死。它特别适用于为研究细胞和分子在转基因小鼠中脑的可塑性响应相关。

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

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Obtaining in vivo human brain tissue volumetrics from MRI is often complicated by various technical and biological issues. These challenges are ...

Minimally Invasive Thumb-sized Pterional Craniotomy for Surgical Clip Ligation of Unruptured Anterior Circulation Aneurysms

微创拇指大小翼点入开颅未破裂前循环动脉瘤的手术夹子结扎

Less invasive surgical approaches for intracranial aneurysm clipping may reduce length of hospital stay, surgical morbidity, treatment cost, and improve ...

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots

封装产热前脂肪细胞移植到脂肪组织仓库

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular ...

A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia

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 ...

Reproducible Motor Deficit Following Aortic Occlusion in a Rat Model Of Spinal Cord Ischemia

主动脉阻塞后脊髓缺血大鼠模型中的可再生电机缺陷

Spinal cord ischemia is a fatal complication following thoracoabdominal aortic aneurysm surgery. Researchers can investigate the strategies for preventing ...

A Minimally Invasive Model to Analyze Endochondral Fracture Healing in Mice Under Standardized Biomechanical Conditions

标准化生物力学条件下小鼠软骨骨折愈合的微创模型分析

Bone healing models are necessary to analyze the complex mechanisms of fracture healing to improve clinical fracture treatment. During the last decade, an ...

Electromagnetic Navigation Transthoracic Nodule Localization for Minimally Invasive Thoracic Surgery

电磁导航经胸结节定位用于微创胸外科手术

The increased use of chest computed tomography (CT) has led to an increased detection of pulmonary nodules requiring diagnostic evaluation and/or ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

重建显微外科缺血再灌注损伤研究的临床前大鼠模型

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Evaluation of Capnography Sampling Line Compatibility and Accuracy when Used with a Portable Capnography Monitor

与便携式刻度学监视器一起使用时，评估卡光采样线兼容性和准确性

Capnography is commonly used to monitor patient&#8217;s ventilatory status. While sidestream capnography has been shown to provide a reliable assessment ...