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2D and 3D Quantitative Analysis of Cell Motility and Cytoskeletal Dynamics

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Cytoskeleton Methods and Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 586))

Summary

2D- and 3D-Dynamic Image Analysis Systems (2D- and 3D-DIAS) for quantitative analysis of cell motility and chemotaxis are described. Particular attention is given to protocols that have proven useful in the quantitation of cell shape changes and pseudopod dynamics during basic cell motility (i.e. crawling in the absence of a chemotactic or other type of extracellular signal) and directed motion. In addition, methods provided, highlight the applicability of this approach to the accurate phenotypic characterizations of cytoskeletal mutations in Dictyostelium discoideum, cytoskeletal alterations in metastatic cells, and cytoskeletal defects in chemotactically defective polymorphonuclear neutrophils.

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References

  1. Ridley, A., Schwartz, M., Burridge, K., Firtel, R., Ginsberg, M., Borisy, G., Parsons, J., and Horwitz, A. (2003) Cell migration: integrating signals from front to back. Science 302, 1704–1709.

    Article  CAS  PubMed  Google Scholar 

  2. Affolter, M. and Weijer, C. (2005) Signaling to cytoskeletal dynamics during chemotaxis. Dev. Cell 9, 19–34.

    Article  CAS  PubMed  Google Scholar 

  3. Janetopoulos, C. and Firtel, R. (2008) Directional sensing during chemotaxis. FEBS Lett. 582, 2075–2085.

    Article  CAS  PubMed  Google Scholar 

  4. Soll, D.R., Wessels, D., Zhang, H., and Heid, P. (2002) A contextual framework for interpreting the roles of proteins in motility and chemotaxis in Dictyostelium discoideum. J. Musc. Res. Cell Motil. 23, 659–672.

    Article  CAS  Google Scholar 

  5. Geiger, J., Wessels, D., and Soll, D.R. (2003) Human polymorphonuclear leukocytes respond to waves of chemoattractant, like Dictyostelium. Cell Motil. Cytoskeleton 56, 27–44.

    Article  PubMed  Google Scholar 

  6. Kolsch, V., Charest, P., and Firtel, R. (2007) The regulation of cell motility and chemotaxis by phospholipid signaling. J. Cell Sci. 121, 551–559.

    Article  Google Scholar 

  7. Wyckoff, J., Wang, W., Lin, E., Wang, Y., Pixley, F., Stanley, E., Graf, T., Pollard, J., Segall, J., and Condeelis, J. (2004) A paracrine loop between tumor cells and macrophages is required for tumor cell migration in mammary tumors. Cancer Res. 64, 7002–7009.

    Article  Google Scholar 

  8. Dumstrei, K., Mennecke, R., and Raz, E. (2004) Signaling pathways controlling primordial germ cell migration in zebrafish. J. Cell Sci. 117, 4787–4795.

    Article  CAS  PubMed  Google Scholar 

  9. Wessels, D., Lusche, D., Kuhl, S., Heid, P., and Soll, D.R. (2007) PTEN plays a role in the suppression of lateral pseudopod formation during Dictyostelium motility and chemotaxis. J. Cell Sci. 120, 2517–2531.

    Article  CAS  PubMed  Google Scholar 

  10. Stepanovic, V., Wessels, D., Goldman, F., Geiger, G., and Soll, D.R. (2004) The chemotaxis defect of Shwachman-Diamond syndrome leukocytes. Cell Motil. Cytoskeleton 57, 158–174.

    Article  CAS  PubMed  Google Scholar 

  11. Williams, R., Boeckeler, K., Graf, R., Taubenberger, A., Li, Z., Isberg, R., Wessels, D., Soll, D.R., Alexander, H., and Alexander, S. (2006) Towards a molecular understanding of human diseases using Dictyostelium discoideum. Trends Mol. Med. 12, 416–422.

    Article  Google Scholar 

  12. Galli, E., Rossi, P., Fiore, L., Perlini, R., Celibon, F., Betti, P., and Businco, L. (1983) IgE levels and PMN chemotaxis in atopic dermatitis. Allergol. Immunopathol. (Madr). 11, 189–194.

    CAS  Google Scholar 

  13. Soll, D.R. (1995) The use of computers in understanding how cells crawl. Int. Rev. Cytol. 163, 43–104.

    Article  CAS  PubMed  Google Scholar 

  14. Soll, D.R. and Voss, E. (1998) Two and three dimensional computer systems for analyzing how cells crawl. In: Motion Analysis of Living Cells (Soll, D.R. and Wessels, D., eds.), Wiley, New York, pp. 25–52.

    Google Scholar 

  15. Soll, D.R., Voss, E., Johnson, O., and Wessels, D.J. (2000) Three-dimensional reconstruction and motion analysis of living crawling cells. Scanning 22, 249–257.

    Article  CAS  PubMed  Google Scholar 

  16. Wessels, D., Kuhl, S., Soll, D.R. (2006) Application of 2D and 3D DIAS to motion analysis of live cells in transmission and confocal microcopy imaging. In: Methods in Molecular Biology (Eichinger, L. and Rivero, F., eds.), Humana, Totowa, NJ, pp. 261–279.

    Google Scholar 

  17. Soll, D.R., Voss, E., Wessels, D., and Kuhl, S. (2007) Computer-assisted systems for dynamic 3D reconstruction and motion analysis of living cells. In: Cell Motility (Shorte, S., ed), Springer, Germany, pp. 365–384.

    Google Scholar 

  18. Wessels, D. and Soll, D.R. (1998) Computer-assisted analysis of cytoskeleton mutants of Dictyostelium. In: Motion Analysis of Living Cells (Soll, D.R. and Wessels, D., eds.), Wiley, New York, pp. 101–140.

    Google Scholar 

  19. Kumar, A., Wessels, D., Daniels, K., Alexander, S., Alexander, H., and Soll, D.R. 2004. Sphingosine-1-phosphate plays a role in the suppression of lateral pseudopod formation during Dictyostelium discoideum cell migration and chemotaxis. Cell Motil. Cytoskeleton 59, 227–241.

    Article  CAS  PubMed  Google Scholar 

  20. Stepanovic, V., Wessels, D., Loomis, W.F., and Soll, D.R. (2005) Adenylyl cyclase plays an intracellular role in the suppression of lateral pseudopod formation during Dictyostelium chemotaxis. Eukaryot. Cell 4, 775–786.

    Article  CAS  PubMed  Google Scholar 

  21. Cocucci, S. and Sussman, M. (1970) RNA in cytoplasmic and nuclear fractions of cellular slime mold amebas. J. Cell Biol. 4, 399–407.

    Article  Google Scholar 

  22. Varnum, B. and Soll, D.R. (1984) Effect of cAMP on single cell motility in Dictyostelium. J. Cell Biol. 99, 1151–1155.

    Article  CAS  PubMed  Google Scholar 

  23. Varnum, B., Edwards, K., and Soll, D.R. (1985) Dictyostelium amoebae alter motility differently in response to increasing versus decreasing temporal gradients of cAMP. J. Cell Biol. 101, 1–5.

    Article  CAS  PubMed  Google Scholar 

  24. Wells, C. and Ridley, A. (2005) Analysis of cell migration using the Dunn chemotaxis chamber and time-lapse microscopy. Methods Mol. Biol. 294, 31–41.

    PubMed  Google Scholar 

  25. Zigmond, S.H. (1978) A new visual assay of leukocyte chemotaxis. In: Leukocyte Chemotaxis: Methods, Physiology and Clinical Implication (Gallin, J.I. and Quie, P.G., eds.), Raven Press, New York, NY, pp. 57–66.

    Google Scholar 

  26. Varnum-Finney, B., Voss, E., and Soll, D.R. (1987) Frequency and orientation of pseudopod formation of Dictyostelium discoideum amoebae chemotaxing in a spatial gradient: further evidence for a temporal mechanism. Cell Motil. Cytoskeleton 8, 18–26.

    Article  CAS  PubMed  Google Scholar 

  27. Shutt, D., Jenkins, L., Carolan, E., Stapleton, J., Daniels, K., Kennedy, R., and Soll, D.R. (1998) T cell syncytia induced by HIV release T cell chemoattractants: demonstration with a newly developed single cell chemotaxis chamber. J. Cell Sci. 111, 99–109.

    CAS  PubMed  Google Scholar 

  28. Varnum, B., Edwards, K., and Soll, D.R. (1986) The developmental regulation of single cell motility in Dictyostelium discoideum. Dev. Biol. 113, 218–227.

    Article  CAS  PubMed  Google Scholar 

  29. Wessels, D., Brincks, R., Kuhl, S., Stepanovic, S., Daniels, K.J., Weeks, G., Lim, C.J., Fuller, D., Loomis, W.F., and Soll, D.R. (2004) RasC plays a selective role in the transduction of temporal gradient information in the cAMP wave of Dictyostelium. Eukaryot. Cell 3, 646–662.

    Article  CAS  PubMed  Google Scholar 

  30. Lokuta, M., Nuzzi, P., and Huttenlocher, A. (2007) Analysis of neutrophil polarization and chemotaxis. Methods Mol. Biol. 412, 211–229.

    Article  CAS  PubMed  Google Scholar 

  31. Sidani, M., Wessels, D., Mouneimne, G., Ghosh, M., Goswami, S., Sarnuebtim C., Wang, W., Kuhl, S., El-Sibai, M., Backer, J., Eddy, R., Soll, D., and Condeelis, J. (2007) Cofilin determines the migration behavior and turning frequency of metastatic cancer cells. J. Cell Biol. 179, 777–791.

    Article  CAS  PubMed  Google Scholar 

  32. Wong, K., Wessels, D., Krob, S., Matveia, A.R., Lin, J.L.-C., Soll, D.R., and Lin, J.J.-C. (2000) Forced expression of dominant-negative chimeric tropomyosin causes abnormal motile behavior during cell division. Cell Motil. Cytoskeleton 45, 121–132.

    Article  CAS  PubMed  Google Scholar 

  33. Li, Y., Wessels, D., Wang, T., Lin, J.L.-C., Soll, D.R., and Lin, J.J-C. (2003) Regulation of caldesmon activity Cdc2 kinase plays an important role in maintaining membrane cortex integrity during cell division. CMLS 60, 198–211.

    Article  CAS  PubMed  Google Scholar 

  34. Volk, P., Heise, C., Hougen, J., Artman, C., Volk, K., Wessels, D., Soll, D., Nauseef, W., Lamb, F., and Moreland, J. (2008) CLC-3 and ICLswell are required for normal neutrophil chemotaxis and shape change. J. Biol. Chem.

    Google Scholar 

  35. Wessels, D., Soll, D.R., Knecht, D., Loomis, W.F., DeLozanne, A., and Spudich, J. (1988) Cell motility and chemotaxis in Dictyostelium amoebae lacking myosin heavy chain. Dev. Biol. 128, 164–177.

    Article  CAS  PubMed  Google Scholar 

  36. Wessels, D., Murray, J., Jung, G., Hammer, J., and Soll, D.R. (1991) Myosin IB null mutant of Dictyostelium exhibits abnormalities in motility. Cell Motil. Cytoskeleton 20, 301–315.

    Article  CAS  PubMed  Google Scholar 

  37. Cox, D., Condeelis, J., Wessels, D., Doll, D., Kern, H., and Knecht, D. (1992) Targeted disruption of the ABP-120 gene leads to cells with altered motility. J. Cell Biol. 116, 943–955.

    Article  CAS  PubMed  Google Scholar 

  38. Titus, M., Wessels, D., Spudich, J., and Soll, D.R. (1992) The unconventional myosin encoded by the myo A gene plays a role in Dictyostelium motility. Mol. Biol. Cell 4, 233–246.

    Google Scholar 

  39. Wessels, D. and Soll, D.R. (1998) Computer-assisted characterization of the behavioral defects of cytoskeletal mutants of Dictyostelium discoideum. In: Motion Analysis of Living Cells (Soll, D.R. and Wessels, D., eds.), Wiley, New York, pp. 101–140.

    Google Scholar 

  40. Wessels, D., Voss, E., Von Bergen, N., Burns, R., Stites, J., and Soll, D.R. (1998) A computer-assisted system for reconstructing and interpreting in three dimensions the relationships of the cell surface, the nucleus and pseudopods in living cells. Cell Motil. Cytoskeleton 41, 225–246.

    Article  CAS  PubMed  Google Scholar 

  41. Heid, P., Voss, E., and Soll, D.R. (2002) 3D-DIASemb: a computer-assisted system for reconstructing and motion analyzing in 4D every cell and nucleus in a developing embryo. Dev. Biol. 245, 329–347.

    Article  CAS  PubMed  Google Scholar 

  42. Pang, K., Lee, E., and Knecht, D. (1998) Use of a fusion protein between GFP and an actin-binding domain to visualize transient filamentous actin structures. Curr. Biol. 8, 405–408.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

The development of 2D- and 3D-DIAS was supported in part by grants HD18577 and AI40040 from the National Institutes of Health and a generous facility grant from the W.M. Keck Foundation.

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Authors and Affiliations

  1. Keck Dynamic Image Analysis Facility, Department of Biological Sciences, University of Iowa, Iowa City, IA, USA

    Deborah Wessels, Spencer Kuhl & David R. Soll

Authors
  1. Deborah Wessels
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  2. Spencer Kuhl
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  3. David R. Soll
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Editors and Affiliations

  1. Brooklyn College, City University of New York, Bedford Ave. 2900, New York, 11210-2889, U.S.A.

    Ray H. Gavin

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Wessels, D., Kuhl, S., Soll, D.R. (2009). 2D and 3D Quantitative Analysis of Cell Motility and Cytoskeletal Dynamics. In: Gavin, R. (eds) Cytoskeleton Methods and Protocols. Methods in Molecular Biology, vol 586. Humana Press. https://doi.org/10.1007/978-1-60761-376-3_18

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  • DOI: https://doi.org/10.1007/978-1-60761-376-3_18

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-375-6

  • Online ISBN: 978-1-60761-376-3

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