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Methods for Investigating Cell Division Mechanisms in C. elegans

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Mitosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2415))

Abstract

The nematode Caenorhabditis elegans is a widely used model organism for the study of mitotic and meiotic cell division. These self-fertilizing worms are particularly advantageous for such studies because they rapidly reproduce (each worm lays ~250 eggs in only 3–4 days) and the cell division machinery is highly conserved between worms and humans. Worms are also genetically tractable and proteins can be readily depleted using RNA interference (RNAi), allowing for the characterization of protein function in vivo. To assess phenotypes, spindles can be directly visualized within the worm using fluorescent protein tags or embryos can be dissected out of the worm and immunostained. A combination of these techniques allows comprehensive characterization of a protein’s function in a relatively short time span. Here, we describe methods for each of these techniques: RNA interference through feeding, in utero live imaging, in utero fixed imaging, and immunofluorescence.

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Acknowledgements

The authors thank Wignall Lab members past and present for their contributions in developing these protocols, especially Gabriel Cavin-Meza, Amanda Davis-Roca, Carissa Heath, Jeremy Hollis, Hannah Horton, Tim Mullen, Christina Muscat, and Michael Tran. This work was supported by American Heart Association Predoctoral Fellowship 17PRE33440016 (to I.D.W.), NIH/NIGMS Molecular Biophysics Training Grant T32GM008382 (to I.D.W.), and NIH R01GM124354 (to S.M.W.). Microscopy was performed at the Biological Imaging Facility at Northwestern University, supported by the Chemistry for Life Processes Institute, the NU Office for Research, the Department of Molecular Biosciences, and the Rice Foundation.

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Authors and Affiliations

  1. Department of Molecular Biosciences, Northwestern University, Evanston, IL, USA

    Ian D. Wolff, Nikita S. Divekar & Sarah M. Wignall

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  1. Ian D. Wolff
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  2. Nikita S. Divekar
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  3. Sarah M. Wignall
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Correspondence to Sarah M. Wignall .

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Editors and Affiliations

  1. The Hormel Institute, University of Minnesota, Austin, MN, USA

    Edward H. Hinchcliffe

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Wolff, I.D., Divekar, N.S., Wignall, S.M. (2022). Methods for Investigating Cell Division Mechanisms in C. elegans. In: Hinchcliffe, E.H. (eds) Mitosis. Methods in Molecular Biology, vol 2415. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1904-9_2

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  • DOI: https://doi.org/10.1007/978-1-0716-1904-9_2

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1903-2

  • Online ISBN: 978-1-0716-1904-9

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