Letter | Published:

Spatiotemporal dynamics of RhoA activity in migrating cells

Nature volume 440, pages 10691072 (20 April 2006) | Download Citation

Subjects

Abstract

Rho family GTPases regulate the actin and adhesion dynamics that control cell migration. Current models postulate that Rac promotes membrane protrusion at the leading edge and that RhoA regulates contractility in the cell body1,2. However, there is evidence that RhoA also regulates membrane protrusion3,4. Here we use a fluorescent biosensor, based on a novel design preserving reversible membrane interactions, to visualize the spatiotemporal dynamics of RhoA activity during cell migration. In randomly migrating cells, RhoA activity is concentrated in a sharp band directly at the edge of protrusions. It is observed sporadically in retracting tails, and is low in the cell body. RhoA activity is also associated with peripheral ruffles and pinocytic vesicles, but not with dorsal ruffles induced by platelet-derived growth factor (PDGF). In contrast to randomly migrating cells, PDGF-induced membrane protrusions have low RhoA activity, potentially because PDGF strongly activates Rac, which has previously been shown to antagonize RhoA activity5,6. Our data therefore show that different extracellular cues induce distinct patterns of RhoA signalling during membrane protrusion.

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Acknowledgements

We are grateful to G. Bokoch, P. Sun, M. Schwartz, R. Tsien, C. Der and E. Sahai for reagents, and to F. Shen for help with image analysis. This work was supported by grants from the Swiss National Science Foundation, Roche Research Foundation, Novartis and Philip Morris to O.P., and from the National Institutes of Health to K.M.H. and R.L.K.

Author information

Author notes

    • Olivier Pertz
    • , Louis Hodgson
    • , Richard L. Klemke
    •  & Klaus M. Hahn

    †Present addresses: University of California at San Diego, Department of Pathology and Moores Cancer Center, Basic Science Building, Room 1040, 9500 Gilman Drive, MC 0612, La Jolla, California 92093, USA (O.P., R.L.K.); University of North Carolina at Chapel Hill, Department of Pharmacology and Lineberger Cancer Center, Chapel Hill, North Carolina 27599, USA (L.H., K.M.H.)

Affiliations

  1. University of North Carolina at Chapel Hill, Department of Pharmacology and Lineberger Cancer Center, Chapel Hill, North Carolina 27599, USA

    • Olivier Pertz
    • , Louis Hodgson
    •  & Klaus M. Hahn
  2. Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA

    • Olivier Pertz
    • , Louis Hodgson
    • , Richard L. Klemke
    •  & Klaus M. Hahn

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Competing interests

K.M.H. is a paid consultant of Panomics Corporation, which markets reagents for high-content microscopy screening.

Corresponding authors

Correspondence to Olivier Pertz or Klaus M. Hahn.

Supplementary information

PDF files

  1. 1.

    Supplementary Notes

    This file contains Supplementary Methods and Supplementary Figures 1–6.

Videos

  1. 1.

    Supplementary Movie 1

    Phase contrast timelapse movie comparing MEFs with or without biosensor expression (induced versus non-induced).

  2. 2.

    Supplementary Movie 2

    Timelapse movie showing YFP and ratio images of a MEF fibroblast randomly migrating on fibronectin (as in Fig. 2a).

  3. 3.

    Supplementary Movie 3

    Timelapse movie showing ratio images of a MEF fibroblast randomly migrating on fibronectin (as in Fig. 2b).

  4. 4.

    Supplementary Movie 4

    Timelapse movie of a peripheral ruffle.

  5. 5.

    Supplementary Movie 5

    Timelapse movie of PDGF-stimulated pinocytosis.

  6. 6.

    Supplementary Movie 6

    Timelapse movie showing robust PDGF stimulation of protrusions in the MEF cells, as shown in Fig.4a.

  7. 7.

    Supplementary Movie 7

    Timelapse movie showing PDGF induction of membrane protrusions in MEF cells expressing T19N DN biosensor.

  8. 8.

    Supplementary Movie 8

    Timelapse movie of a MEF migrating out of a wounded monolayer, then stimulated with PDGF.

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DOI

https://doi.org/10.1038/nature04665

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