Isolating vessels from the mouse brain for gene expression analysis using laser capture microdissection
Isolating vessels from the mouse brain for gene expression analysis using laser capture microdissection
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DOI：
10.1016/s1385-299x(02)00147-2
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发表时间：
2002-06-01
期刊：
影响因子：
--
通讯作者：
Hunt, NH
中科院分区：
文献类型：
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作者：
Ball, HJ;McParland, B;Hunt, NH
对基因表达的研究通常检查从构成组织的不同细胞类型中提取的RNA池。我们开发了一种从脑中分离血管的方法，以了解脑型疟疾发病过程中血管的变化。血管是通过用碱性磷酸酶底物孵育小鼠大脑的切片来显示的。通过激光捕获显微解剖收集血管，并通过测量细胞特异性标志物的表达来监测其特异性。从捕获的血管中提取的RNA高度富含与内皮细胞和周细胞相关的基因的mRNA。吲哚胺2，3-双加氧酶mRNA的测定表明，有可能检测到由于疟疾感染而导致的基因表达的变化，这种变化专门发生在血管内。激光捕获显微切割可以用来研究血脑屏障上基因表达的变化。皇冠版权所有(C)2002由爱思唯尔科学公司出版。保留所有权利。
Studies of gene expression often examine a pool of RNA extracted from the diverse cell types making up a tissue. We have developed a method for isolating vessels from the brain in order to understand the changes occurring in the vessels during the pathogenesis of cerebral malaria. Vessels were visualised by incubating sections of mouse brain with a substrate for alkaline phosphatase. Vessels were collected by laser capture microdissection and the specificity was monitored by measuring the expression of cell-specific markers. RNA from the captured vessels was highly enriched in mRNA for genes associated with endothelial cells and pericytes. Measurement of indoleamine 2,3-dioxygenase mRNA indicated it was possible to detect changes in gene expression, due to malaria infection, occurring specifically within the vessels. Laser capture microdissection can be used to study changes in gene expression occurring at the blood-brain barrier. Crown copyright (C) 2002 Published by Elsevier Science B.V. All rights reserved.