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

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Cilia

Part of the book series: Methods in Molecular Biology ((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.

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Acknowledgments

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

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Authors and Affiliations

  1. Human Genetics Institute of New Jersey, Rutgers, The State University of New Jersey, 145 Bevier Rd., Piscataway, NJ, 08854, USA

    Robert O’Hagan Ph.D. & Maureen M. Barr

Authors
  1. Robert O’Hagan Ph.D.
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  2. Maureen M. Barr
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Corresponding author

Correspondence to Robert O’Hagan Ph.D. .

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Editors and Affiliations

  1. Albert Einstein College of Medicine, Yeshiva University, Bronx, New York, USA

    Peter Satir

  2. Department of Biology, University of Copenhagen, Copenhagen, Denmark

    Søren Tvorup Christensen

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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: Satir, P., Christensen, S. (eds) Cilia. Methods in Molecular Biology, vol 1454. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3789-9_8

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  • DOI: https://doi.org/10.1007/978-1-4939-3789-9_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3787-5

  • Online ISBN: 978-1-4939-3789-9

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