Abstract
Cytosolic fluid dynamics have been implicated in cell motility1,2,3,4,5 because of the hydrodynamic forces they induce and because of their influence on transport of components of the actin machinery to the leading edge. To investigate the existence and the direction of fluid flow in rapidly moving cells, we introduced inert quantum dots into the lamellipodia of fish epithelial keratocytes and analysed their distribution and motion. Our results indicate that fluid flow is directed from the cell body towards the leading edge in the cell frame of reference, at about 40% of cell speed. We propose that this forward-directed flow is driven by increased hydrostatic pressure generated at the rear of the cell by myosin contraction, and show that inhibition of myosin II activity by blebbistatin reverses the direction of fluid flow and leads to a decrease in keratocyte speed. We present a physical model for fluid pressure and flow in moving cells that quantitatively accounts for our experimental data.
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References
Charras, G., Yarrow, J., Horton, M., Mahadevan, L. & Mitchison, T. Non-equilibration of hydrostatic pressure in blebbing cells. Nature 435, 365–369 (2005).
Oster, G. F. & Perelson, A. S. The physics of cell motility. J. Cell Sci. Suppl. 8, 35–54 (1987).
Rubinstein, B., Jacobson, K. & Mogilner, A. Multiscale two-dimensional modeling of a motile simple-shaped cell. Multiscale Modeling Simul. 3, 413–439 (2005).
Saadoun, S., Papadopoulos, M. C., Hara-Chikuma, M. & Verkman, A. S. Impairment of angiogenesis and cell migration by targeted aquaporin-1 gene disruption. Nature 434, 786–792 (2005).
Loitto, V. M., Karlsson, T. & Magnusson, K. E. Water flux in cell motility: expanding the mechanisms of membrane protrusion. Cell Motil. Cytoskel. 66, 237–247 (2009).
Loitto, V. M., Forslund, T., Sundqvist, T., Magnusson, K. E. & Gustafsson, M. Neutrophil leukocyte motility requires directed water influx. J. Leukoc. Biol. 71, 212–222 (2002).
Hu, J. & Verkman, A. S. Increased migration and metastatic potential of tumor cells expressing aquaporin water channels. FASEB J. 20, 1892–1894 (2006).
Zicha, D. et al. Rapid actin transport during cell protrusion. Science 300, 142–145 (2003).
Allen, R. D. & Allen, N. S. Cytoplasmic streaming in amoeboid movement. Ann. Rev. Biophys. Bioeng. 7, 469–495 (1978).
Abraham, V. C., Krishnamurthi, V., Taylor, D. L. & Lanni, F. The actin-based nanomachine at the leading edge of migrating cells. Biophys. J. 77, 1721–1732 (1999).
Svitkina, T. M., Verkhovsky, A. B., McQuade, K. M. & Borisy, G. G. Analysis of the actin-myosin II system in fish epidermal keratocytes: mechanism of cell body translocation. J. Cell Biol. 139, 397–415 (1997).
Euteneuer, U. & Schliwa, M. Persistent, directional motility of cells and cytoplasmic fragments in the absence of microtubules. Nature 310, 58–61 (1984).
Lee, J., Ishihara, A., Theriot, J. A. & Jacobson, K. Principles of locomotion for simple-shaped cells. Nature 362, 167–171 (1993).
Dubertret, B. et al. In vivo imaging of quantum dots encapsulated in phospholipid micelles. Science 298, 1759–1762 (2002).
Valentine, M. T. et al. Colloid surface chemistry critically affects multiple particle tracking measurements of biomaterials. Biophys. J. 86, 4004–4014 (2004).
Lacayo, C. I. et al. Emergence of large-scale cell morphology and movement from local actin filament growth dynamics. PLoS Biol. 5, e233 (2007).
Keren, K. et al. Mechanism of shape determination in motile cells. Nature 453, 475–480 (2008).
Qian, H., Sheetz, M. P. & Elson, E. L. Single particle tracking. Analysis of diffusion and flow in two-dimensional systems. Biophys. J. 60, 910–921 (1991).
Luby-Phelps, K. Cytoarchitecture and physical properties of cytoplasm: volume, viscosity, diffusion, intracellular surface area. Int. Rev. Cytol. 192, 189–221 (2000).
Theriot, J. A. & Mitchison, T. J. Actin microfilament dynamics in locomoting cells. Nature 352, 126–131 (1991).
Wilson, C. A. Large scale coordination of actin meshwork flow in rapidly moving cells. PhD Thesis, Stanford Univ. (2006).
Vallotton, P., Danuser, G., Bohnet, S., Meister, J.-J. & Verkhovsky, A. B. Tracking Retrograde Flow in Keratocytes: News from the Front. Mol. Biol. Cell 16, 1223–1231 (2005).
Olbrich, K., Rawicz, W., Needham, D. & Evans, E. Water permeability and mechanical strength of polyunsaturated lipid bilayers. Biophys. J. 79, 321–327 (2000).
Lee, J., Ishihara, A., Oxford, G., Johnson, B. & Jacobson, K. Regulation of cell movement is mediated by stretch-activated calcium channels. Nature 400, 382–386 (1999).
Straight, A. F. et al. Dissecting temporal and spatial control of cytokinesis with a myosin II Inhibitor. Science 299, 1743–1747 (2003).
Iwasaki, T. & Wang, Y. L. Cytoplasmic force gradient in migrating adhesive cells. Biophys. J. 94, L35–37 (2008).
Galbraith, C. G. & Sheetz, M. P. Keratocytes pull with similar forces on their dorsal and ventral surfaces. J. Cell Biol. 147, 1313–1324 (1999).
Oliver, T., Dembo, M. & Jacobson, K. Separation of propulsive and adhesive traction stresses in locomoting keratocytes. J. Cell Biol. 145, 589–604 (1999).
Prass, M., Jacobson, K., Mogilner, A. & Radmacher, M. Direct measurement of the lamellipodial protrusive force in a migrating cell. J. Cell Biol. 174, 767–772 (2006).
Anderson, K. & Cross, R. Contact dynamics during keratocyte motility. Curr. Biol. 10, 253–260 (2000).
Yam, P. T. et al. Actin–myosin network reorganization breaks symmetry at the cell rear to spontaneously initiate polarized cell motility. J. Cell Biol. 178, 1207–1221 (2007).
Kolega, J. Phototoxicity and photoinactivation of blebbistatin in UV and visible light. Biochem. Biophys. Res. Comm. 320, 1020–1025 (2004).
Sbalzarini, I. F. & Koumoutsakos, P. Feature point tracking and trajectory analysis for video imaging in cell biology. J. Struct. Biol. 151, 182–195 (2005).
Slepchenko, B. M., Schaff, J. C., Macara, I. & Loew, L. M. Quantitative cell biology with the Virtual Cell. Trends Cell Biol. 13, 570–576 (2003).
Acknowledgements
We thank Theresa Harper and Marcel Bruchez from Quantum Dot Corporation (Molecular Probes, Invitrogen) for providing the quantum dot probes; W.E. Moerner for advice on the single-particle tracking experiments; Paul Wiseman, Ben Hebert and Lia Gracey for their contributions to the initial phase of this project; Michael Saxton and Cyrus Wilson for useful discussions; and Boris Slepchenko for help with Virtual Cell. K.K. was supported by a Damon Runyon Postdoctoral Fellowship, a Horev fellowship from the Technion, an Allon Fellowship from the Israel Council for Higher Education, and by grants from the Morasha Program of the Israel Science Foundation, the Converging Technologies Program of The Israel Council for Higher Education, the Wolfson Foundation, and a European Research Council Starting Grant. A.M. was supported by grants from the National Institutes of Health and the National Science Foundation. J.A.T. was supported by grants from the National Institutes of Health, the American Heart Association and the Howard Hughes Medical Institute. P.T.Y. was supported by a Howard Hughes Medical Institute Predoctoral Fellowship and Stanford Graduate Fellowship.
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K.K. and J.A.T. conceived and designed the experiments; K.K. performed the experiments and analysed the data; A.M. and K.K. developed the model; P.T.Y. made the initial observation of enhancement of large probes at the leading edge; A.K. contributed to the single-particle tracking experiments; K.K., A.M., P.T.Y. and J.A.T. discussed the results and wrote the paper.
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Keren, K., Yam, P., Kinkhabwala, A. et al. Intracellular fluid flow in rapidly moving cells. Nat Cell Biol 11, 1219–1224 (2009). https://doi.org/10.1038/ncb1965
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DOI: https://doi.org/10.1038/ncb1965
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