Probing the CNS microvascular endothelium by immune-guided laser-capture microdissection coupled to quantitative RT-PCR.
Probing the CNS microvascular endothelium by immune-guided laser-capture microdissection coupled to quantitative RT-PCR.
复制标题
通过免疫引导激光捕获显微切割结合定量 RT-PCR 探测中枢神经系统微血管内皮。
DOI：
10.1007/978-1-61779-163-5_32
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Pachter,JoelS
中科院分区：
文献类型：
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作者：
Murugesan,Nivetha;Macdonald,Jennifer;Ge,Shujun;Pachter,JoelS
激光捕获显微解剖（LCM）允许检索不同群体的细胞从他们的紧密周围的邻居在原位。因此，LCM对于研究沿中枢神经系统（CNS）微血管内皮的基因表达非常有利，中枢神经系统（CNS）微血管内皮是一种表现出相当大的节段和区域异质性的组织。结合CNS微血管内皮细胞的免疫组织化学染色和血管周围星形细胞或平滑肌细胞的免疫荧光染色，免疫引导的LCM（免疫-LCM）可以与下游的qRT-PCR相结合，探测健康和疾病期间内皮沿CNS微血管树的表达变化。免疫- lcm /qRT-PCR已被用于突出中枢神经系统微血管各部分对血脑屏障（BBB）的贡献，并可用于检查病理过程中血脑屏障基因表达的变化。
Laser-capture microdissection (LCM) allows for retrieval of distinct populations of cells from their closely surrounding neighbors in situ. As such, LCM is highly advantageous for investigating gene expression along the central nervous system (CNS) microvascular endothelium, a tissue that shows both considerable segmental and regional heterogeneity. Combining immunohistochemical staining of CNS microvascular endothelial cells with immunofluorescent staining of perivascular astrocytes or smooth muscle cells, immune-guided LCM,immuno-LCM, may be coupled to downstream qRT-PCR to probe varied expression of the endothelium along the CNS microvascular tree during health and disease. Immuno-LCM/qRT-PCR has been used to highlight contributions of the respective segments of the CNS microvasculature to the blood–brain barrier (BBB), and can be employed to examine changes in BBB gene expression during pathology.