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Analysis of neuronal gene expression with laser capture microdissection
Author(s) -
Vincent Valerie A.M.,
DeVoss Jason J.,
Ryan Heather S.,
Murphy Greer M.
Publication year - 2002
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.10329
Subject(s) - laser capture microdissection , gene expression , microdissection , biology , gene , hippocampal formation , cell type , cell , reverse transcriptase , polymerase chain reaction , microbiology and biotechnology , neuroscience , genetics
The brain is a heterogeneous tissue in which the numbers of neurons, glia, and other cell types vary among anatomic regions. Gene expression studies performed on brain homogenates yield results reflecting mRNA abundance in a mixture of cell types. Therefore, a method for quantifying gene expression in individual cell populations would be useful. Laser capture microdissection (LCM) is a new technique for obtaining pure populations of cells from heterogeneous tissues. Most studies thus far have used LCM to detect DNA sequences. We developed a method to quantify gene expression in hippocampal neurons from mouse brain using LCM and real‐time reverse transcriptase‐polymerase chain reaction (RT‐PCR). This method was optimized to permit histochemical or immunocytochemical visualization of nerve cells during LCM while minimizing RNA degradation. As an example, gene expression was quantified in hippocampal neurons from the Tg2576 mouse model for Alzheimer's disease. © 2002 Wiley‐Liss, Inc.