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Profiling Axonal mRNA Transport

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RNA Detection and Visualization

Part of the book series: Methods in Molecular Biology ((MIMB,volume 714))

Abstract

The importance of mRNA localization and localized protein synthesis to spatially modulate protein levels in distinct subcellular domains has increasingly been recognized in recent years. Axonal and dendritic processes of neurons represent separate functional domains of the cell that have shown the capacity to autonomously respond to extracellular stimuli through localized protein synthesis. With the vast distance often separating distal axons and dendrites from the neuronal cell body, these processes have provided an appealing and useful model system to study the mechanisms that drive mRNA localization and regulate localized mRNA translation. Here, we discuss the methodologies that have been used to isolate neuronal processes to purity, and provide an in-depth method for using a modified Boyden chamber to isolate axons from adult dorsal root ganglion neurons for analyses of axonal mRNA content. We further show how this method can be utilized to identify specific mRNAs whose transport into axons is altered in response to extracellular stimuli, providing a means to begin to understand how axonal protein synthesis contributes to the proper function of the neuron.

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Acknowledgements

The methods presented here were developed using funds from the National Institutes of Health (R01-NS041596 and R01-NS049041 to JLT and K99-NR010797 to DEW).

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Authors and Affiliations

  1. Department of Biology, Drexel University, Philadelphia, PA, USA

    Jeffery L. Twiss

Authors
  1. Dianna E. Willis
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  2. Jeffery L. Twiss
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Correspondence to Jeffery L. Twiss .

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Editors and Affiliations

  1. , Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, 76100, Israel

    Jeffrey E. Gerst

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Willis, D.E., Twiss, J.L. (2011). Profiling Axonal mRNA Transport. In: Gerst, J. (eds) RNA Detection and Visualization. Methods in Molecular Biology, vol 714. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-005-8_21

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  • DOI: https://doi.org/10.1007/978-1-61779-005-8_21

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-004-1

  • Online ISBN: 978-1-61779-005-8

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