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Fibronectin organization under and near cells

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Abstract

Polymerization of soluble fibronectin molecules results in fibres that are visible as networks using fluorescently labelled fibronectin protomers or by antibody labelling. Displacement of fibres composed of modified protomers in living cells provides information regarding matrix structure, organization, and movement. A static analysis of fibronectin structures and patterns of organization provide insight into their reorganization during adhesion and motility. Confocal microscopy and atomic force microscopy (AFM) reveal fibronectin-containing networks aligned in arrays perpendicular to the retracting cell edge and in apparently disordered networks of fibres under the cell. The change in patterns suggests a reorganization of fibronectin from disordered arrays used for adhesion into ordered arrays during movement of the cell. Comparison of confocal images with corresponding AFM images confirms that the fibres left on the surface as the cell moves away do contain fibronectin. The orientation of these fibres relative to the tail (uropod) and the receding edges of the cell leads us to propose that cells generate a force on the fibres that exceeds the adhesion force of the fibres to the surface causing them to pull fibronectin fibres into straight arrays. However, when the fibres are parallel to the direction of pull, the fibres remain attached to the surface. The data supports the hypothesis that disorganized, linear fibres are the product of Fn polymerization induced by the cell beneath it and serve to adhere the cell to the substrate as the cell spreads, whereas arrays of fibres found outside the cell are formed as existing fibrils and reorganize during cell motility.

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Abbreviations

Fn:

Fibronectin

ECM:

Extracellular matrix

AFM:

Atomic Force Microscopy

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Acknowledgements

The authors would like to thank Dr. Niki Boyd for fruitful discussions, the Petersen and Norton group members, and the Sandig lab in the Department of Anatomy and Cell Biology for their technical assistance. We would also like to acknowledge financial support from NSERC for strategic grants and CFI for infrastructure grants to NOP and PRN as well as OGSST and NSERC for postgraduate scholarships for KLD.

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Author notes

  1. Kathy L. De Jong

    Present address: Xerox Research Centre Canada, Mississauga, ON, Canada, L5K 2L1

Authors and Affiliations

  1. Department of Chemistry, University of Western Ontario, London, ON, Canada, N6A 5B7

    Kathy L. De Jong, Heather C. MacLeod, Peter R. Norton & Nils O. Petersen

  2. National Institute of Nanotechnology, 11421 Saskatchewan Drive, Edmonton, AB, Canada, T6G 2M9

    Nils O. Petersen

Authors
  1. Kathy L. De Jong
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  2. Heather C. MacLeod
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Correspondence to Nils O. Petersen.

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De Jong, K.L., MacLeod, H.C., Norton, P.R. et al. Fibronectin organization under and near cells. Eur Biophys J 35, 695–708 (2006). https://doi.org/10.1007/s00249-006-0081-7

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  • DOI: https://doi.org/10.1007/s00249-006-0081-7

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