Kymographic analysis of transport in an individual neuronal sensory cilium in caenorhabditis elegans

Robert O’Hagan, Maureen M. Barr

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

Abstract

Intraflagellar Transport (IFT) is driven by molecular motors that travel upon microtubule-based ciliary axonemes. In the single-celled alga Chlamydomonas reinhardtii, movement of a single anterograde IFT motor, heterotrimeric kinesin-II, is required to generate two identical motile flagella. The function of this canonical anterograde IFT motor is conserved among all eukaryotes, yet multicellular organisms can generate cilia of diverse structures and functions, ranging from simple threadlike non-motile primary cilia to the elaborate cilia that make up rod and cone photoreceptors in the retina. An emerging theme is that additional molecular motors modulate the canonical IFT machinery to give rise to differing ciliary morphologies. Therefore, a complete understanding of the trafficking of ciliary receptors, as well as the biogenesis, maintenance, specialization, and function of cilia, requires the characterization of motor molecules. Here, we describe in detail our method for measuring the motility of proteins in cilia or dendrites of C. elegans male-specific CEM ciliated sensory neurons using time-lapse microscopy and kymography of green fluorescent protein (GFP)-tagged motors, receptors, and cargos. We describe, as a specific example, OSM- 3::GFP puncta moving in cilia, but also include (Fig. 1) with settings that have worked well for us measuring movement of heterotrimeric kinesin-II, IFT particles, and the polycystin TRP channel PKD-2.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages107-122
Number of pages16
DOIs
StatePublished - 1 Jan 2016

Publication series

NameMethods in Molecular Biology
Volume1454
ISSN (Print)1064-3745

Keywords

  • Caenorhabditis elegans
  • Cell biology
  • In vivo
  • KLP-6
  • Kinesin-2
  • Kinesin-3
  • Kymograph
  • Male
  • OSM-3
  • PKD- 2
  • Polycystins
  • Sensory non-motile cilia

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    O’Hagan, R., & Barr, M. M. (2016). Kymographic analysis of transport in an individual neuronal sensory cilium in caenorhabditis elegans. In Methods in Molecular Biology (pp. 107-122). (Methods in Molecular Biology; Vol. 1454). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3789-9_8