期刊
JOURNAL OF NEUROSCIENCE METHODS
卷 208, 期 2, 页码 108-113出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jneumeth.2012.04.014
关键词
Astrocyte; Oligodendrocyte; Microglia; RNA; Laser capture microdissection; Post-mortem human brain
资金
- Multiple Sclerosis Society of Great Britain and Northern Ireland, registered charity [207495]
- Medical Research Council, UK
- Biogen Idec
- MRC [MR/J004308/1] Funding Source: UKRI
- Medical Research Council [MR/J004308/1] Funding Source: researchfish
Isolating individual populations of cells from post-mortem (PM) central nervous system (CNS) tissue for transcriptomic analysis will provide important insights into the pathogenesis of neurodegenerative diseases. To date, research on individual CNS cell populations has been hindered by the availability of suitable PM material, unreliable sample preparation and difficulties obtaining individual cell populations. In this paper we report how rapid immunohistochemistry combined with laser capture microdissection (immuno-LCM) enables the isolation of specific cell populations from PM CNS tissue, thereby enabling the RNA profile of these individual cell types to be investigated. Specifically, we detail methods for isolating enriched glial populations (astrocytes, oligodendrocytes and microglia) and confirm this cell enrichment by polymerase chain reaction (PCR). In addition, the study details the numbers of each glial population required to obtain 50 ng RNA, a suitable amount of starting material required to carry out microarray analysis that potentially may identify alterations of cell-specific genes and pathways associated with a range of neurodegenerative disorders. (C) 2012 Elsevier B.V. All rights reserved.
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