Identification of process-localized mRNAs from cultured rodent hippocampal neurons

Michael M. Poon, Sang-Hyun Choi, Christina A.M. Jamieson, Daniel H. Geschwind, Kelsey C. Martin

Research output: Contribution to journalArticle

145 Citations (Scopus)

Abstract

The regulated translation of localized mRNAs in neurons provides a mechanism for spatially restricting gene expression in a synapse-specific manner. To identify the population of mRNAs present in distal neuronal processes of rodent hippocampal neurons, we grew neurons on polycarbonate filters etched with 3 μm pores. Although the neuronal cell bodies remained on the top surface of the filters, dendrites, axons, and glial processes penetrated through the pores to grow along the bottom surface of the membrane where they could be mechanically separated from cell bodies. Quantitative PCR and immunochemical analyses of the process preparation revealed that it was remarkably free of somatic contamination. Microarray analysis of RNA isolated from the processes identified over 100 potentially localized mRNAs. In situ hybridization studies of 19 of these transcripts confirmed that all 19 were present in dendrites, validating the utility of this approach for identifying dendritically localized transcripts. Many of the identified mRNAs encoded components of the translational machinery and several were associated with the RNA-binding protein Staufen. These findings indicate that there is a rich repertoire of mRNAs whose translation can be locally regulated and support the emerging idea that local protein synthesis serves to boost the translational capacity of synapses.

Original languageEnglish
Pages (from-to)13390-13399
Number of pages10
JournalJournal of Neuroscience
Volume26
Issue number51
DOIs
Publication statusPublished - 2006 Dec 20

Fingerprint

Rodentia
polycarbonate
Protein Biosynthesis
Dendrites
Neurons
Synapses
Messenger RNA
RNA-Binding Proteins
Microarray Analysis
Neuroglia
In Situ Hybridization
Axons
RNA
Gene Expression
Polymerase Chain Reaction
Membranes
Population
Proteins
Cell Body

Keywords

  • Dendrite
  • Hippocampus
  • in situ hybridization
  • Microarray
  • mRNA localization
  • Protein synthesis
  • Synaptic plasticity

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Identification of process-localized mRNAs from cultured rodent hippocampal neurons. / Poon, Michael M.; Choi, Sang-Hyun; Jamieson, Christina A.M.; Geschwind, Daniel H.; Martin, Kelsey C.

In: Journal of Neuroscience, Vol. 26, No. 51, 20.12.2006, p. 13390-13399.

Research output: Contribution to journalArticle

Poon, Michael M. ; Choi, Sang-Hyun ; Jamieson, Christina A.M. ; Geschwind, Daniel H. ; Martin, Kelsey C. / Identification of process-localized mRNAs from cultured rodent hippocampal neurons. In: Journal of Neuroscience. 2006 ; Vol. 26, No. 51. pp. 13390-13399.
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