Article | Published:

A soft cortex is essential for asymmetric spindle positioning in mouse oocytes

Nature Cell Biology volume 15, pages 958966 (2013) | Download Citation

Abstract

At mitosis onset, cortical tension increases and cells round up, ensuring correct spindle morphogenesis and orientation. Thus, cortical tension sets up the geometric requirements of cell division. On the contrary, cortical tension decreases during meiotic divisions in mouse oocytes, a puzzling observation because oocytes are round cells, stable in shape, that actively position their spindles. We investigated the pathway leading to reduction in cortical tension and its significance for spindle positioning. We document a previously uncharacterized Arp2/3-dependent thickening of the cortical F-actin essential for first meiotic spindle migration to the cortex. Using micropipette aspiration, we show that cortical tension decreases during meiosis I, resulting from myosin-II exclusion from the cortex, and that cortical F-actin thickening promotes cortical plasticity. These events soften and relax the cortex. They are triggered by the Mos–MAPK pathway and coordinated temporally. Artificial cortex stiffening and theoretical modelling demonstrate that a soft cortex is essential for meiotic spindle positioning.

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Acknowledgements

We thank T. Pollard (Yale University, USA) for providing the CK666 and F. Perez (Curie Institute, France) for providing SF9-expressing plasmids. We thank J. Teillon for helping set up the photoactivation experiments. We thank C. Klein for his help with the analysis of the photoactivation data. We also thank A. Roux, M. Piel, A. Echard, A. Gautreau, E. Derivery and R. Li for helpful discussions. This work was supported by grants from the Ligue Nationale Contre le Cancer (EL2009-EL2012/LNCC/MHV) and from the Agence Nationale pour la Recherche (ANR-08-BLAN-0136-01 to MHV and ANR-08-BLAN-0012-12 to CS). A. Chaigne is a recipient of a fellowship from the Ecole Normale Supérieure (ENS) Paris. C. Campillo acknowledges financial support from the Association pour la Recherche contre le Cancer (ARC). N. S. Gov wishes to thank the Mayent-Rothschild Foundation for the Visiting Professor grant at the Curie Institute.

Author information

Author notes

    • Clément Campillo
    •  & Pierre Nassoy

    Present addresses: Institut Lumière Matière, UMR5306 CNRS/UCBL, Villeurbanne 69622, France (C.C.); LP2N, Institut d’Optique, CNRS UMR 5298 & Université de Bordeaux 1, F-33405 Talence, France (P.N.)

    • Nir S. Gov
    •  & Raphaël Voituriez

    These authors contributed equally to this work

Affiliations

  1. CIRB, Collège de France, and CNRS-UMR7241 and INSERM-U1050, Equipe Labellisée Ligue Contre le Cancer, Paris F-75005, France

    • Agathe Chaigne
    • , Jessica Azoury
    • , Claudia Umaña-Diaz
    • , Maria Almonacid
    • , Isabelle Queguiner
    • , Marie-Hélène Verlhac
    •  & Marie-Emilie Terret
  2. Memolife Laboratory of Excellence and Paris Science Lettre, Paris F-75005, France

    • Agathe Chaigne
    • , Jessica Azoury
    • , Claudia Umaña-Diaz
    • , Maria Almonacid
    • , Isabelle Queguiner
    • , Marie-Hélène Verlhac
    •  & Marie-Emilie Terret
  3. Institut Curie, Centre de Recherche, Laboratoire Physico-Chimie, Paris F-75248, France

    • Clément Campillo
    • , Pierre Nassoy
    •  & Cécile Sykes
  4. Department of Chemical Physics, Weizmann Institute of Science, Rehovot 76100, Israel

    • Nir S. Gov
  5. UMR7600-CNRS/UPMC, 4 Place Jussieu, Paris F-75005, France

    • Raphaël Voituriez
  6. CNRS-UMR168, Paris F-75248, France

    • Pierre Nassoy
    •  & Cécile Sykes
  7. UPMC, 4 Place Jussieu, Paris F-75248, France

    • Pierre Nassoy
    •  & Cécile Sykes

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Contributions

A.C., M-H.V. and M-E.T. designed the experiments, interpreted the results and wrote the manuscript. A.C., C.U-D. and M.E.T. carried out most of the experiments. J.A. performed the preliminary observations that started the project. M.A. assisted in the photoactivation experiments. I.Q. assisted in genotyping the animals. A.C. and C.C. carried out the micropipette experiments. C.C., P.N. and C.S. designed the micropipette experiments and interpreted the results. N.S.G. and R.V. designed the physical model. M.E.T. and M.H.V. conceived and supervised the project.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Marie-Hélène Verlhac or Marie-Emilie Terret.

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Videos

  1. 1.

    Time-lapse movie showing a WT oocyte expressing GFP–UtrCH (grey) and histone–RFP (red).

    Frames were 1 h apart. The movie starts at NEBD and stops at NEBD+7 h. One Z plane is shown.

  2. 2.

    Time-lapse movie showing a WT oocyte expressing GFP–UtrCH.

    Frames were 5 s apart. The movie starts at NEBD+7 h and lasts 5 min. One Z plane is shown.

  3. 3.

    Pseudocolored time-lapse movies showing WT NEBD+6 h oocytes expressing PA–GFP–UtrCH (upper cells) and PA–GFP (lower cells) photoactivated in the subcortex (cells on the left) or in the cytoplasm (cells on the right).

    Black, lowest intensity; white, highest intensity. Frames were 100 milliseconds apart. The movies start at NEBD+6 h and last 25 seconds. One Z plane is shown.

  4. 4.

    13-second movie showing micropipette aspiration of a WT NEBD+1h mouse oocyte.

    Note that the plasma membrane does not detach from the cytoplasm, as cytoplasmic particles can be seen flowing into the micropipette.

  5. 5.

    31-second movies showing the relaxation after suction of a NEBD+6 h mos-/- oocyte (left cell, elastic behaviour) and a NEBD+6 h WT oocyte (right cell, plastic behaviour).

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DOI

https://doi.org/10.1038/ncb2799

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