Journal
JOURNAL OF NEUROSCIENCE METHODS
Volume 143, Issue 2, Pages 95-106Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jneumeth.2004.09.023
Keywords
microarray; laser capture microdissection; dopamine neurons
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Funding
- NIDA NIH HHS [R41 DA016406] Funding Source: Medline
- NIEHS NIH HHS [P30 ES06639] Funding Source: Medline
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The cellular heterogeneity of brain tissue presents a challenge to gene expression profiling of specific neuronal cell types. The present study employed a fluorescent neural tracer to specifically label midbrain dopamine neurons and non-dopamine cortical neurons. The labeled cells were then used to visually guide harvesting of the cells by laser capture microdissection (LCM). RNA extracted from the two populations of harvested cells was then amplified, labeled and co-hybridized to high density cDNA microarrays for two-color differential expression profiling. Many of the genes most highly enriched in the dopamine neurons were found to be genes previously known to define the dopamine neuronal phenotype. However, results from the microarray were only partially validated by quantitative RT-PCR analysis. The results indicate that LCM harvesting of specific neuronal phenotypes can be effectively guided in a complex cellular environment by specific pre-labeling of the target cell populations and underlie the importance of independent validation of microarray results. (c) 2004 Elsevier B.V. All rights reserved.
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