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Myofibril contraction and crosslinking drive nuclear movement to the periphery of skeletal muscle

Nature Cell Biology volume 19, pages 11891201 (2017) | Download Citation

Abstract

Nuclear movements are important for multiple cellular functions, and are driven by polarized forces generated by motor proteins and the cytoskeleton. During skeletal myofibre formation or regeneration, nuclei move from the centre to the periphery of the myofibre for proper muscle function. Centrally located nuclei are also found in different muscle disorders. Using theoretical and experimental approaches, we demonstrate that nuclear movement to the periphery of myofibres is mediated by centripetal forces around the nucleus. These forces arise from myofibril contraction and crosslinking that ‘zip’ around the nucleus in combination with tight regulation of nuclear stiffness by lamin A/C. In addition, an Arp2/3 complex containing Arpc5L together with γ-actin is required to organize desmin to crosslink myofibrils for nuclear movement. Our work reveals that centripetal forces exerted by myofibrils squeeze the nucleus to the periphery of myofibres.

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Acknowledgements

We thank M.-H. Verlhac, M. Dias, J. Pinto, G. Gundersen and S. Tapscott for comments on the manuscript. We thank the Gomes Laboratory for discussions. This work was supported by the European Research Council (E.R.G.), EMBO installation (E.R.G.), the myograd PhD programme (W.R.), AIM France (W.R., B.C., E.R.G.), LISBOA-01-0145-FEDER-007391 co-funded by FEDER through POR Lisboa 2020—Programa Operacional Regional de Lisboa, do PORTUGAL 2020 (E.R.G.), and Fundação para a Ciência e a Tecnologia (E.R.G.). M.W. and J.V.G.A. were supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001209), the UK Medical Research Council (FC001209) and the Wellcome Trust (FC001209), as well as by postdoctoral fellowships from FRQS (Fonds de recherche du Québec—Santé), EMBO and the Canadian Institutes of Health Research (CIHR) to J.V.G.A.

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Affiliations

  1. Sorbonne Universités, UPMC Univ Paris 06, INSERM UMRS974, CNRS FRE3617, Center for Research in Myology, GH Pitié-Salpêtrière, 47 Boulevard de l’Hôpital, 75013 Paris, France

    • William Roman
    • , Bruno Cadot
    •  & Edgar R. Gomes
  2. Centre de Référence de Pathologie Neuromusculaire Paris-Est, Institut de Myologie, GHU La Pitié-Salpêtrière, Assistance Publique-Hôpitaux de Paris, 75013 Paris, France

    • William Roman
    • , Bruno Cadot
    •  & Edgar R. Gomes
  3. Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Avenida Professor Egas Moniz, 1649-028 Lisboa, Portugal

    • William Roman
    • , João P. Martins
    • , Filomena A. Carvalho
    • , Nuno C. Santos
    •  & Edgar R. Gomes
  4. Laboratoire de Physique Théorique de la Matière Condensée, CNRS UMR 7600, Université Pierre et Marie Curie, 75013 Paris, France

    • Raphael Voituriez
  5. Laboratoire Jean Perrin, CNRS FRE 3231, Université Pierre et Marie Curie, 75013 Paris, France

    • Raphael Voituriez
  6. Cellular Signalling and Cytoskeletal Function Lab, The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK

    • Jasmine V. G. Abella
    •  & Michael Way

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Contributions

W.R. carried out experiments and analysed data; J.M. performed lamin- and desmin-related experiments; F.A.C. and N.C.S. carried out AFM experiments. W.R., B.C. and E.R.G. conceived and designed experiments; J.V.G.A. and M.W. provided Arp2/3-related unpublished tools; W.R., R.V. and B.C. designed and executed the physical model; W.R. and E.R.G. wrote the manuscript with assistance from other authors; all authors participated in the critical review and revision of the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Edgar R. Gomes.

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Videos

  1. 1.

    Centrally located nucleus surrounded by myofibrils.

    Surface three-dimensional rendering of a time-lapse movie (also presented in Fig S3C) depicting a centrally located nucleus surrounded by myofibrils bundles while moving longitudinally in a 4-day myofiber expressing YFP-α-actinin (green, myofibrils) and H2B-iRFP (red, nucleus).

  2. 2.

    Nuclear movement to the periphery.

    Time-lapse movie of a 5-day myofiber depicting peripheral movement of a nucleus (red, H2B-iRFP) through myofibrils (green, α-YFP-actinin) (also presented in Fig 3A). Top left: top view of surface three-dimensional rendering. Middle left: side view of surface three-dimensional rendering. Bottom left: 2D view from the top of the central plane. Top right: nucleus alone viewed from 90° rotation surface three-dimensional rendering. Bottom right: nucleus with transparent myofibers from 90° rotation surface three-dimensional rendering.

  3. 3.

    Nuclear wrinkles.

    Surface three-dimensional rendering of the same nucleus (red, H2B-iRFP) with more (wrinkled- right) or less (unwrinkled- left) wrinkles due to variations in myofibril tension rotating 360° along its longitudinal axis (also presented in Supplementary Fig. 5A).

  4. 4.

    Optogenetically controlled myofiber contraction.

    Time-lapse movie of 3.5-day myofibers transfected with ChR2-GFP (left) and untransfected (right) (also presented in Supplementary Fig. 5B). Blue light is shone at the rhythm of the ‘eye of the tiger’ by Survivors to show controllability of contraction by light. We advise appropriate volume during video playback.

  5. 5.

    Nuclear movement to the periphery induced by myofiber contraction.

    Time-lapse movie of a 3.5-day myofiber transfected with ChR2-GFP (green) and H2B-iRFP (red) and untransfected (also presented in Fig. 4f). Top: ChR2-GFP (green) and H2B-iRFP (red) channels. Bottom: H2B-iRFP (red) and bright-field (grey) channels. Blue light is shone 6 pulses per hour to induce contraction. 00:00 refers to 3.5 days.

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DOI

https://doi.org/10.1038/ncb3605

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