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Basic mechanism of three-dimensional collagen fibre transport by fibroblasts

Nature Cell Biology volume 7, pages 157–164 (2005)Cite this article

Abstract

Collagen remodelling by fibroblasts has a crucial role in organizing tissue structures that are essential to motility during wound repair, development and regulation of cell growth. However, the mechanism of collagen fibre movement in three-dimensional (3D) matrices is not understood. Here, we show that fibroblast lamellipodia extend along held collagen fibres, bind, and retract them in a 'hand-over-hand' cycle, involving α2β1 integrin. Wild-type fibroblasts move collagen fibres three to four times farther per cycle than fibroblasts lacking myosin II-B (myosin II-B−/−). Similarly, myosin II-B−/− fibroblasts contract 3D collagen gels threefold less than controls. On two-dimensional (2D) substrates, however, rates of collagen bead and cell movement are not affected by loss of myosin II-B. Green fluorescent protein (GFP)-tagged myosin II-B, but not II-A, restores normal function in knockout cells and localizes to cell processes, whereas myosin II-A is more centrally located. Additionally, GFP–myosin II-B moves out to the periphery and back during hand-over-hand fibre movement, whereas on 2D collagen, myosin II-B is more centrally distributed. Thus, we suggest that cyclic myosin II-B assembly and contraction in lamellipodia power 3D fibre movements.

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Figure 1: Fibroblasts transport individual collagen fibres in 'hand-over-hand' events of extension and contraction.
Figure 2: NMHC II-B deletion causes defects in rearward transport of collagen fibres.
Figure 3: Loss of NMHC II-B causes a decrease in 3D collagen gel contraction.
Figure 4: NMHC II-B is enriched in regions of cell–collagen attachment.
Figure 5: NMHC II-B is present in 3D but not in 2D extending/retracting lamellipodia.

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Acknowledgements

We thank J. Holmes for his help with the 3D collagen gel experiments; M. Erne and M. Barr for their data analysis; O. Rossier, A. Kostić, H.-G. Döbereiner and G. Giannone for their careful review and comments on the manuscript; and K. De Vos and members of the Sheetz Laboratory for their stimulating conversations. This work is supported by an NIH grant to M.P.S.

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Authors and Affiliations

  1. Department of Biological Sciences, Columbia University, New York, 10027, NY, USA

    Adam S. Meshel & Michael P. Sheetz

  2. Laboratory of Molecular Cardiology, National Heart, Lung, and Blood Institute, NIH, Bethesada, 20892, MD, USA

    Qize Wei & Robert S. Adelstein

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  1. Adam S. Meshel
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  2. Qize Wei
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Correspondence to Michael P. Sheetz.

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Meshel, A., Wei, Q., Adelstein, R. et al. Basic mechanism of three-dimensional collagen fibre transport by fibroblasts. Nat Cell Biol 7, 157–164 (2005). https://doi.org/10.1038/ncb1216

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