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 journalArticlepeer-review

    160 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

    Keywords

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

    ASJC Scopus subject areas

    • Neuroscience(all)

    Fingerprint

    Dive into the research topics of 'Identification of process-localized mRNAs from cultured rodent hippocampal neurons'. Together they form a unique fingerprint.

    Cite this