Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical Lesion Model for Mouse Stroke Studies
Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical Lesion Model for Mouse Stroke Studies
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DOI：
10.3791/50370
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发表时间：
2013-06-01
影响因子：
1.2
通讯作者：
Tomasi, Simone
中科院分区：
文献类型：
--
作者：
Labat-gest, Vivien;Tomasi, Simone
光血栓卒中模型旨在通过对先前注射的光敏染料进行光激活，在给定的皮质区域内诱导缺血性损伤。光照后，染料被激活并产生单线态氧，破坏内皮细胞膜的成分，随后发生血小板聚集和血栓形成，最终决定局部血液流动的中断。这一方法最初由Rosenblum和El-Sabban于1977年提出，后来Watson于1985年在大鼠大脑中进行了改进，并为目前的模型奠定了基础。此外，转基因小鼠品系的增加也进一步提高了人们对光血栓模型的兴趣。简而言之，一种光敏染料(孟加拉玫瑰)被注射到腹膜内并进入血流。当被冷光源照射时，染料被激活，并诱导内皮损伤，导致血小板激活和血栓形成，导致局部血流中断。该光源可以应用在完整的头骨上，而不需要开颅手术，这允许以可重现和非侵入性的方式瞄准任何感兴趣的皮质区域。然后缝合小鼠，让它苏醒。用氯化三苯基四氮唑或甲酚紫染色可以快速完成对缺血损伤的评估。这项技术可以产生小范围和边界清楚的梗塞，这对于精确的细胞特征或功能研究是非常有利的。此外，它特别适合于研究转基因小鼠大脑可塑性背后的细胞和分子反应。
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.