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External forces control mitotic spindle positioning

Abstract

The response of cells to forces is essential for tissue morphogenesis and homeostasis. This response has been extensively investigated in interphase cells, but it remains unclear how forces affect dividing cells. We used a combination of micro-manipulation tools on human dividing cells to address the role of physical parameters of the micro-environment in controlling the cell division axis, a key element of tissue morphogenesis. We found that forces applied on the cell body direct spindle orientation during mitosis. We further show that external constraints induce a polarization of dynamic subcortical actin structures that correlate with spindle movements. We propose that cells divide according to cues provided by their mechanical micro-environment, aligning daughter cells with the external force field.

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Figure 1: Retraction fibre distribution dictates mitotic spindle orientation.
Figure 2: Retraction fibres exert strong forces on the mitotic cell body.
Figure 3: Stretching retraction fibres induces spindle rotation.
Figure 4: Adhesion geometry can bias dynamic subcortical actin structures in mitotic cells.
Figure 5: Quantification of subcortical actin polarization for different adhesion geometries.
Figure 6: Polarization of dynamic subcortical actin structures persists when astral microtubules are depolymerized and the spindle is misoriented.
Figure 7: Mitotic spindle rotation and movement strongly correlate with polarization of dynamic subcortical actin structures.
Figure 8: Subcortical actin structures exert pulling forces on the mitotic spindle.

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Acknowledgements

We are grateful to J. Colombelli for helpful discussions and preliminary experiments on retraction fibre cutting; L. Mahadevan for helpful discussions concerning mechanical cortex anisotropy; D. Gerlich (ETH, Zurich, Switzerland) and R. Tsien (UCSD, San Diego, USA) for the pIRES-puro3–MyrPalm–GFP plasmid; V. Doye (IJM, Paris, France) for the pIRES-neo–histone2B–mCherry plasmid; W. Bement (University of Wisconsin, Madison, USA) for the GFP–Utr-CH plasmid; R. Wedlich-Soldner (IMPRS, Martinsried, Germany) and G. Montagnac (Institut Curie, Paris, France) for the Lifeact–mCherry plasmid; M. Heuze (Institut Curie, Paris, France) and A. M. Lennon-Dumesnil (Institut Curie, Paris, France) for the Lifeact–mCherry lentivirus; V. Fraisier, the Nikon Imaging Center and the PICT–IBiSA of the Institut Curie for technical support in microscopy; J. Boulanger for image treatment using ndsafir; and Z. Maciarowski, C. Guérin and A. Viguier for FACS sorting of stable cell lines. We thank M. Thery and J. Aubertin for helpful discussions throughout the course of this work and A. W. Murray, E. Paluch, A. M. Lennon-Dumesnil, A. Taddei, M. Thery, S. Misery-Lenkei and members of the Piel laboratory for critical reading of the manuscript. This work was supported by the Centre National de la Recherche Scientifique, the Institut Curie and by ANR (ANR-06-PCVI-0010) and HFSP grants to M.P. J.F. was supported by pre-doctoral fellowships from Boehringer Ingelheim Fonds and the Association pour la Recherche sur le Cancer.

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J.F. designed, carried out and analysed most experiments and wrote the article, N.C. carried out most cell stretching experiments as well as experiments shown in Fig. 2f and Supplementary Fig. S3, T.B. carried out and analysed optical trap experiments (Fig. 2c–e), A.B. carried out some cell stretching experiments, M.C. carried out the experiments shown in Fig. 2f and Supplementary Fig. S3, A.A. developed the method to produce micropatterns on stretchable substrates, M.B. contributed ideas, discussion and supervised part of the work of J.F., C.S. contributed ideas and discussion and supervised the work on optical trap experiments, L.F. carried out laser ablation experiments (Fig. 1), D.C. set up the cell stretching device, supervised the work of A.B. and contributed ideas and discussion, and M.P. supervised the work, carried out experiments and wrote the paper.

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Correspondence to Matthieu Piel.

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Fink, J., Carpi, N., Betz, T. et al. External forces control mitotic spindle positioning. Nat Cell Biol 13, 771–778 (2011). https://doi.org/10.1038/ncb2269

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