Measuring rates of intraflagellar transport along Caenorhabditis elegans sensory cilia using fluorescence microscopy

Ingrid Brust-Mascher, Guangshuo Ou, Jonathan M. Scholey

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

Abstract

Intraflagellar transport (IFT), the kinesin-2 and IFT-dynein-dependent bidirectional movement of multisubunit protein complexes called IFT-particles and associated cargo molecules along ciliary axonemes, is thought to be essential for the assembly and maintenance of virtually all eukaryotic cilia and flagella. Transport assays that allow measurements of the rates of movement of specific, fluorescently tagged, functional components of the IFT machinery, including motors, IFT particle subunits, and putative cargo, were first developed in Caenorhabditis elegans sensory cilia, and they have proved to be an important and valuable tool for dissecting the molecular mechanisms of IFT. We describe how these transport assays are performed in our laboratory and summarize the information that has been obtained by using them concerning the mechanisms of action and regulation of the motors that drive IFT, the composition and organization of the IFT-particles, and the identification of IFT-dynein subunits and ciliary tubulin isotypes as likely cargo proteins of kinesin-2-driven anterograde IFT.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
Pages285-304
Number of pages20
Volume524
DOIs
StatePublished - 2013

Publication series

NameMethods in Enzymology
Volume524
ISSN (Print)00766879
ISSN (Electronic)15577988

Keywords

  • Cilium assembly
  • Fluorescence microscopy
  • Fluorescence recovery after photobleaching
  • In vivo motility assays
  • Intraflagellar transport assays
  • Kinesin-2
  • Mathematical modeling
  • Tubulin

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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  • Cite this

    Brust-Mascher, I., Ou, G., & Scholey, J. M. (2013). Measuring rates of intraflagellar transport along Caenorhabditis elegans sensory cilia using fluorescence microscopy. In Methods in Enzymology (Vol. 524, pp. 285-304). (Methods in Enzymology; Vol. 524). https://doi.org/10.1016/B978-0-12-397945-2.00016-0