A new method for determining blood-brain barrier integrity based on intracardiac perfusion of an Evans Blue-Hoechst cocktail
A new method for determining blood-brain barrier integrity based on intracardiac perfusion of an Evans Blue-Hoechst cocktail
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DOI：
10.1016/j.jneumeth.2008.06.025
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发表时间：
2008-09-15
影响因子：
3
通讯作者：
Pelegri, Carme
中科院分区：
文献类型：
--
作者：
del Valle, Jaume;Camins, Antoni;Pelegri, Carme
描述了一种确定血脑屏障（BBB）改变的大脑区域的新方法。在该方法中，小鼠心内灌注伊文思蓝 (EB) 和添加在标准甲醛固定溶液中的 Hoechst 示踪剂。在动物大脑中诱导的局部冷冻损伤产生脑部水肿且血脑屏障受损后，对这种鸡尾酒方法进行了测试。然后将结果与心内灌注前静脉内和腹膜内施用示踪剂进行比较。使用鸡尾酒法时，红色 EB 荧光定位冷冻损伤的大脑区域，而 Hoechst 示踪剂则对同一区域的细胞核进行染色。在没有功能性血脑屏障的脉络丛和室周器官中也可以观察到 EB 和 Hoechst 荧光。与使用该示踪剂的传统方法相比，该混合物在 BBB 破坏区域中提供了更强烈的 EB 染色。当以混合物形式施用时，赫斯特示踪剂也更有用，因为当静脉内施用时，它会染色所有脑核。鸡尾酒法允许对脑切片进行免疫染色，使研究人员能够表征和分析存在 BBB 紊乱的区域的结构和细胞变化。因此，免疫组织化学已用于确定出现在病灶周围边缘的强 EB 荧光细胞的性质，这些细胞在此被鉴定为神经元细胞。 (C) 2008 Elsevier B.V. 保留所有权利。
A new method for determining brain regions with blood-brain barrier (BBB) alterations is described. In this method, mice were perfused intracardially with Evans Blue (EB) and Hoechst tracers added in a standard formaldehyde fixative solution. This cocktail method was tested after a localized cryolesion induced in the brain had produced an edematous brain region with disrupted BBB in the animals. The results were then compared with the intravenous and intraperitoneal administration of the tracers prior to intracardiac perfusion. When using the cocktail method, red EB fluorescence locates the cryoinjured brain region while the Hoechst tracer stains the nuclei in that same region. EB and Hoechst fluorescence can also be observed in the choroid plexus and circumventricular organs, where there is no functional BBB. The cocktail gives more intense EB staining in zones of disrupted BBB than that given by traditional methods which use this tracer. The Hoechst tracer is also more useful when administered in the cocktail, since when administrated intravenously it stains all the brain nuclei. The cocktail method permits the immunostaining of brain sections, enabling researchers to characterize and analyze structural and cellular changes in regions where BBB disturbances are present. Thus, immunohistochemistry has been used here to determine the nature of intense EB fluorescent cells that appear in the perilesional rim, which were identified here as neuronal cells. (C) 2008 Elsevier B.V. All rights reserved.