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Cilia pp 107-122 | Cite as

Kymographic Analysis of Transport in an Individual Neuronal Sensory Cilium in Caenorhabditis elegans

  • Robert O’HaganEmail author
  • Maureen M. Barr
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1454)

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.

Key words

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

Notes

Acknowledgments

The authors were supported by NJCSCR Grant CSCR15IRG014 (R.O.) and NIH Grants DK059418 and DK074746 (M.B.).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Human Genetics Institute of New Jersey, RutgersThe State University of New JerseyPiscatawayUSA

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