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  Global transcriptome analysis of genetically identified neurons in the adult cortex

Rossner, M. J., Hirrlinger, J., Wichert, S. P., Boehm, C., Newrzella, D., Hiemisch, H., et al. (2006). Global transcriptome analysis of genetically identified neurons in the adult cortex. Journal of Neuroscience, 26(39), 9956-9966. doi:10.1523/JNEUROSCI.0468-06.2006.

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 Creators:
Rossner, Moritz J.1, Author              
Hirrlinger, Johannes1, Author              
Wichert, Sven P.1, Author              
Boehm, Christine, Author
Newrzella, Dieter, Author
Hiemisch, Holger, Author
Eisenhardt, Gisela, Author
Stuenkel, Carolin1, Author              
Ahsen, Oliver von, Author
Nave, Klaus-Armin1, Author              
Affiliations:
1Neurogenetics, Max Planck Institute of Experimental Medicine, Max Planck Society, ou_2173664              

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Free keywords: cortex; cerebellum; microarrays; transcriptome; microdissection; GFP GENE-EXPRESSION ANALYSIS; CENTRAL-NERVOUS-SYSTEM; TRANSGENIC MICE; REGULATED GENES; BRAIN-FUNCTION; IN-SITU; MOUSE; MICROARRAYS; HYBRIDIZATION; GLYCOLYSIS
 Abstract: The enormous cellular complexity of the brain is a major obstacle for gene expression profiling of neurological disease models, because physiologically relevant changes of transcription in a specific neuronal subset are likely to be lost in the presence of other neurons and glia. We solved this problem in transgenic mice by labeling genetically defined cells with a nuclear variant of GFP. When combined with laser-directed microdissection, intact RNA from unfixed, freeze-dried sections can be isolated, which is a prerequisite for high-quality global transcriptome analysis. Here, we compared gene expression profiles between pyramidal motor neurons and pyramidal somatosensory neurons captured from layer V of the adult neocortex. One striking feature of motor neurons is the elevated expression of ribosomal genes and genes involved in ATP synthesis. This suggests a molecular adaptation of the upper motor neurons to longer axonal projections and higher electrical activity. These molecular signatures were not detected when cortical layers and microareas were analyzed in toto. Additionally, we used microarrays to determine the global mRNA expression profiles of microdissected Purkinje cells and cellularly complex cerebellar cortex microregions. In summary, our analysis shows that cellularly complex targets lead to averaged gene expression profiles that lack substantial amounts of cell type-specific information. Thus, cell type-restricted sampling strategies are mandatory in the CNS. The combined use of a genetic label with laser-microdissection offers an unbiased approach to map patterns of gene expression onto practically any cell type of the brain.

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Language(s): eng - English
 Dates: 2006-09
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 292154
ISI: 000240825700015
ISI: 000240825700015
DOI: 10.1523/JNEUROSCI.0468-06.2006
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Title: Journal of Neuroscience
  Alternative Title : J. Neurosci.
Source Genre: Journal
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Pages: - Volume / Issue: 26 (39) Sequence Number: - Start / End Page: 9956 - 9966 Identifier: ISSN: 0270-6474