Mammalian growth cone turning assays identify distinct cell signalling mechanisms that underlie axon growth, guidance and regeneration

Andrew J. Murray, Andrew G. Peace, Steven J. Tucker, Derryck A. Shewan

Research output: Chapter in Book/Report/Conference proceedingConference contribution

5 Citations (Scopus)

Abstract

The cell signalling mechanisms underlying mammalian central nervous system axon growth and guidance change during development, such that axons that establish appropriate connectivity in the embryo fail to regenerate after injury to the adult nervous system. The growth cone turning assay has been used in Xenopus neurons to elucidate mechanisms of axon guidance during development. Here, we describe how we have adapted this assay for rat dorsal root ganglion neurons to study the influence of extracellular secreted factors causing growth cone attraction and repulsion. Additionally, we describe how this method can be combined with small interfering RNA and cDNA transfections to manipulate protein expression in growth cones, and fluorescence resonance energy transfer to monitor the activity of signalling pathways in live neurons. This assay provides the unique ability to manipulate and visualise the internal status of growth cone signalling whilst challenged with extracellular chemotropic signalling molecules, and can be used to develop strategies to promote axon regeneration in the mature mammalian central nervous system.
Original languageEnglish
Title of host publicationMethods in Molecular Biology
Subtitle of host publicationChapter 15 Neurotrophic factors
PublisherHumana Press
Pages167-178
Number of pages12
Volume846
ISBN (Electronic)978-1-61779-536-7
ISBN (Print)978-1-61779-535-0
DOIs
Publication statusPublished - 2012

Publication series

NameMethods in Molecular Biology: Methods and Protocols
PublisherHumana Press
Volume846
ISSN (Print)1064-3745

Fingerprint

Growth Cones
Regeneration
Growth
Neurons
Axons
Central Nervous System
Fluorescence Resonance Energy Transfer
Spinal Ganglia
Xenopus
Nervous System
Small Interfering RNA
Transfection
Embryonic Structures
Complementary DNA
Axon Guidance
Wounds and Injuries
Proteins

Keywords

  • axon guidance
  • regeneration
  • growth cone
  • turning assay
  • fluorescence resonance energy transfer
  • cell signalling
  • cAMP

Cite this

Murray, A. J., Peace, A. G., Tucker, S. J., & Shewan, D. A. (2012). Mammalian growth cone turning assays identify distinct cell signalling mechanisms that underlie axon growth, guidance and regeneration. In Methods in Molecular Biology: Chapter 15 Neurotrophic factors (Vol. 846, pp. 167-178). (Methods in Molecular Biology: Methods and Protocols; Vol. 846). Humana Press. https://doi.org/10.1007/978-1-61779-536-7_15

Mammalian growth cone turning assays identify distinct cell signalling mechanisms that underlie axon growth, guidance and regeneration. / Murray, Andrew J.; Peace, Andrew G.; Tucker, Steven J.; Shewan, Derryck A.

Methods in Molecular Biology: Chapter 15 Neurotrophic factors. Vol. 846 Humana Press, 2012. p. 167-178 (Methods in Molecular Biology: Methods and Protocols; Vol. 846).

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Murray, AJ, Peace, AG, Tucker, SJ & Shewan, DA 2012, Mammalian growth cone turning assays identify distinct cell signalling mechanisms that underlie axon growth, guidance and regeneration. in Methods in Molecular Biology: Chapter 15 Neurotrophic factors. vol. 846, Methods in Molecular Biology: Methods and Protocols, vol. 846, Humana Press, pp. 167-178. https://doi.org/10.1007/978-1-61779-536-7_15
Murray AJ, Peace AG, Tucker SJ, Shewan DA. Mammalian growth cone turning assays identify distinct cell signalling mechanisms that underlie axon growth, guidance and regeneration. In Methods in Molecular Biology: Chapter 15 Neurotrophic factors. Vol. 846. Humana Press. 2012. p. 167-178. (Methods in Molecular Biology: Methods and Protocols). https://doi.org/10.1007/978-1-61779-536-7_15
Murray, Andrew J. ; Peace, Andrew G. ; Tucker, Steven J. ; Shewan, Derryck A. / Mammalian growth cone turning assays identify distinct cell signalling mechanisms that underlie axon growth, guidance and regeneration. Methods in Molecular Biology: Chapter 15 Neurotrophic factors. Vol. 846 Humana Press, 2012. pp. 167-178 (Methods in Molecular Biology: Methods and Protocols).
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