通过免疫引导激光捕获显微切割结合定量逆转录聚合酶链反应探测中枢神经系统微血管内皮细胞。

Probing the CNS microvascular endothelium by immune-guided laser-capture microdissection coupled to quantitative RT-PCR.

作者信息

Murugesan Nivetha, Macdonald Jennifer, Ge Shujun, Pachter Joel S

机构信息

Blood-Brain Barrier Laboratory, Department of Cell Biology, University of Connecticut Health Center, Farmington, CT, USA.

出版信息

Methods Mol Biol. 2011;755:385-94. doi: 10.1007/978-1-61779-163-5_32.

DOI:10.1007/978-1-61779-163-5_32

PMID:21761321

Abstract

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.

摘要

激光捕获显微切割（LCM）能够在原位从其紧密相邻的细胞中分离出不同的细胞群体。因此，LCM对于研究中枢神经系统（CNS）微血管内皮细胞的基因表达具有高度优势，该组织表现出显著的节段性和区域异质性。将CNS微血管内皮细胞的免疫组织化学染色与血管周围星形胶质细胞或平滑肌细胞的免疫荧光染色相结合，免疫引导激光捕获显微切割（免疫LCM）可与下游定量逆转录聚合酶链反应（qRT-PCR）联用，以探究健康和疾病状态下CNS微血管树沿线内皮细胞的不同表达。免疫LCM/qRT-PCR已被用于突出CNS微血管各节段对血脑屏障（BBB）的贡献，并可用于检查病理过程中BBB基因表达的变化。