The use of fluorescence resonance energy transfer (FRET) to measure axon growth and guidance-related intracellular signalling in live dorsal root ganglia neuronal growth cones

Steven John Tucker

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The measurement of signalling by traditional methods in primary neuronal cultures is often limited by cell numbers within the culture and restricted division among these cells. Further limitations are seen with modern fluorescent imaging techniques on account of difficulties with transfection of these cell types. Here, we describe successful transfection of dorsal root ganglion (DRG) primary neuronal cultures with cDNA encoded fluorescence resonance energy transfer (FRET) probes for various signalling moieties, and subsequent measurement of FRET as an index of signalling within these cells. Furthermore, these measurements were made within live neuronal growth cones, which are thin, fragile, and dynamic structures central to axonal growth, repair, and regeneration. This provides novel, physiological insight into the signalling processes driving these axonal behaviors.
Original languageEnglish
Title of host publicationAxon Growth and Regeneration
Subtitle of host publicationMethods and Protocols
EditorsAndrew J Murray
PublisherHumana Press
Pages29-40
Number of pages12
ISBN (Electronic)978-1-4939-0777-9
ISBN (Print)978-1-4939-0776-2
DOIs
Publication statusPublished - 2014

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
Volume1162
ISSN (Electronic)1064-3745

Keywords

  • fluorescence resonance energy transfer
  • growth cones
  • intracellular signalling
  • DRG neurons
  • neuronal signalling
  • axon growth
  • axon guidance
  • live cell imaging

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    Tucker, S. J. (2014). The use of fluorescence resonance energy transfer (FRET) to measure axon growth and guidance-related intracellular signalling in live dorsal root ganglia neuronal growth cones. In A. J. Murray (Ed.), Axon Growth and Regeneration: Methods and Protocols (pp. 29-40). (Methods in Molecular Biology; Vol. 1162). Humana Press. https://doi.org/10.1007/978-1-4939-0777-9