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Cytoskeleton Dynamics pp 297–325Cite as

Using the Four-Cell C. elegans Embryo to Study Contractile Ring Dynamics During Cytokinesis

Part of the Methods in Molecular Biology book series (MIMB,volume 2101)

Abstract

Cytokinesis is the process that completes cell division by partitioning the contents of the mother cell between the two daughter cells. It involves the highly regulated assembly and constriction of an actomyosin contractile ring, whose function is to pinch the mother cell in two. Research on the contractile ring has particularly focused on the signaling mechanisms that dictate when and where the ring is formed. In vivo studies of ring constriction are however scarce and its mechanistic understanding is therefore limited. Here we present several experimental approaches for monitoring ring constriction in vivo, using the four-cell C. elegans embryo as model. These approaches allow for the ring to be perturbed only after it forms and include the combination of live imaging with acute drug treatments, temperature-sensitive mutants and rapid temperature shifts, as well as laser microsurgery. In addition, we explain how to combine these with RNAi-mediated depletion of specific components of the cytokinetic machinery.

Key words

  • C. elegans
  • Cytokinesis
  • Contractile ring
  • Embryonic cell divisions
  • Acute drug treatments
  • Temperature-sensitive mutations
  • RNAi-mediated depletion
  • Laser microsurgery

Fung Yi Chan and Ana Marta Silva are first co-authors.

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  • DOI: 10.1007/978-1-0716-0219-5_18
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Acknowledgments

Research in our group receives funding from the European Research Council under the European Union’s Horizon 2020 research and innovation programme to AXC (grant agreement 640553—ACTOMYO) and from Norte-01-0145-FEDER-000029—Advancing Cancer Research: from basic knowledge to application, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (FEDER). AXC has a Principal Investigator position from FCT (CEECIND/01967/2017). FYC and AMS hold FCT junior researcher positions DL 57/2016/CP1355/CT0013 and DL 57/2016/CP1355/CT0017, respectively. The authors declare no competing financial interests.

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Authors and Affiliations

  1. Instituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, Porto, Portugal

    Fung Yi Chan, Ana Marta Silva & Ana Xavier Carvalho

  2. Instituto de Biologia Molecular e Celular (IBMC), Universidade do Porto, Porto, Portugal

    Fung Yi Chan, Ana Marta Silva & Ana Xavier Carvalho

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  1. Fung Yi Chan
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  2. Ana Marta Silva
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  3. Ana Xavier Carvalho
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Correspondence to Ana Xavier Carvalho .

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  1. i3S University of Porto, Porto, Portugal

    Helder Maiato

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Chan, F.Y., Silva, A.M., Carvalho, A.X. (2020). Using the Four-Cell C. elegans Embryo to Study Contractile Ring Dynamics During Cytokinesis. In: Maiato, H. (eds) Cytoskeleton Dynamics. Methods in Molecular Biology, vol 2101. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0219-5_18

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  • DOI: https://doi.org/10.1007/978-1-0716-0219-5_18

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