Abstract
Cell migration is required for a wide variety of processes from bacteria seeking for food to correct patterning of neuronal networks. The ability to sense external cues is critical for cells to get directions and reach their goals. So far, studies on chemotaxis have mainly focused their attention on individual cells and therefore available tools are designed to monitor cell behavior at the single cell level. However, as collective cell migration is now widely accepted as a main mode of cell migration from development to cancer, the question of how chemotaxis is achieved has also to be asked on a bigger scale. Here, we present two chemotaxis assays suitable for single cells, cell sheets, and cell explants. Using a simple combination of heparin-coated beads and high vacuum silicone grease, these techniques can be adapted to a wide variety of culture conditions. They allow time-lapse study, high-resolution microscopy, and can be set up at no extra cost.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Serafini, T., Colamarino, S. A., Leonardo, E. D., Wang, H., Beddington, R., Skarnes, W. C., and Tessier-Lavigne, M. (1996) Netrin-1 is required for commissural axon guidance in the developing vertebrate nervous system, Cell 87, 1001–1014.
Tessier-Lavigne, M., and Goodman, C. S. (1996) The molecular biology of axon guidance, Science 274, 1123–1133.
Ming, G. L., Song, H. J., Berninger, B., Holt, C. E., Tessier-Lavigne, M., and Poo, M. M. (1997) cAMP-dependent growth cone guidance by netrin-1, Neuron 19, 1225–1235.
Song, H. J., Ming, G. L., and Poo, M. M. (1997) cAMP-induced switching in turning direction of nerve growth cones, Nature 388, 275–279.
Fernandis, A. Z., and Ganju, R. K. (2001) Slit: a roadblock for chemotaxis, Sci STKE 2001, pe1.
Boldajipour, B., Mahabaleshwar, H., Kardash, E., Reichman-Fried, M., Blaser, H., Minina, S., Wilson, D., Xu, Q., and Raz, E. (2008) Control of chemokine-guided cell migration by ligand sequestration, Cell 132, 463–473.
Doitsidou, M., Reichman-Fried, M., Stebler, J., Koprunner, M., Dorries, J., Meyer, D., Esguerra, C. V., Leung, T., and Raz, E. (2002) Guidance of primordial germ cell migration by the chemokine SDF-1, Cell 111, 647–659.
Richardson, B. E., and Lehmann, R. (2010) Mechanisms guiding primordial germ cell migration: strategies from different organisms, Nat Rev Mol Cell Biol 11, 37–49.
Chen, A., Ganor, Y., Rahimipour, S., Ben-Aroya, N., Koch, Y., and Levite, M. (2002) The neuropeptides GnRH-II and GnRH-I are produced by human T cells and trigger laminin receptor gene expression, adhesion, chemotaxis and homing to specific organs, Nat Med 8, 1421–1426.
Butcher, E. C., and Picker, L. J. (1996) Lymphocyte homing and homeostasis, Science 272, 60–66.
Le, Y., Zhou, Y., Iribarren, P., and Wang, J. (2004) Chemokines and chemokine receptors: their manifold roles in homeostasis and disease, Cell Mol Immunol 1, 95–104.
Pals, S. T., de Gorter, D. J., and Spaargaren, M. (2007) Lymphoma dissemination: the other face of lymphocyte homing, Blood 110, 3102–3111.
Boyden, S. (1962) The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes, J Exp Med 115, 453–466.
Lauffenburger, D., Rothman, C., and Zigmond, S. H. (1983) Measurement of leukocyte motility and chemotaxis parameters with a linear under-agarose migration assay, J Immunol 131, 940–947.
Zicha, D., Dunn, G. A., and Brown, A. F. (1991) A new direct-viewing chemotaxis chamber, J Cell Sci 99 (Pt 4), 769–775.
Gundersen, R. W., and Barrett, J. N. (1979) Neuronal chemotaxis: chick dorsal-root axons turn toward high concentrations of nerve growth factor, Science 206, 1079–1080.
Bianco, A., Poukkula, M., Cliffe, A., Mathieu, J., Luque, C. M., Fulga, T. A., and Rorth, P. (2007) Two distinct modes of guidance signalling during collective migration of border cells, Nature 448, 362–365.
Deisboeck, T. S., and Couzin, I. D. (2009) Collective behavior in cancer cell populations, Bioessays 31, 190–197.
Friedl, P., and Gilmour, D. (2009) Collective cell migration in morphogenesis, regeneration and cancer, Nat Rev Mol Cell Biol 10, 445–457.
Rorth, P. (2009) Collective cell migration, Annu Rev Cell Dev Biol 25, 407–429.
Theveneau, E., Marchant, L., Kuriyama, S., Gull, M., Moepps, B., Parsons, M., and Mayor, R. (2010) Collective chemotaxis requires contact-dependent cell polarity, Dev Cell 19, 39–53.
Zigmond, S. H., and Hirsch, J. G. (1973) Leukocyte locomotion and chemotaxis. New methods for evaluation, and demonstration of a cell-derived chemotactic factor, J Exp Med 137, 387–410.
Orr, W., and Ward, P. A. (1978) Quantitation of leukotaxis in agarose by three different methods, J Immunol Methods 20, 95–107.
Pujic, Z., Giacomantonio, C. E., Unni, D., Rosoff, W. J., and Goodhill, G. J. (2008) Analysis of the growth cone turning assay for studying axon guidance, J Neurosci Methods 170, 220–228.
Xu, X., Brzostowski, J. A., and Jin, T. (2009) Monitoring dynamic GPCR signaling events using fluorescence microscopy, FRET imaging, and single-molecule imaging, Methods Mol Biol 571, 371–383.
Hynes, R. O. (2009) The extracellular matrix: not just pretty fibrils, Science 326, 1216–1219.
He, L., and Niemeyer, B. (2003) A novel correlation for protein diffusion coefficients based on molecular weight and radius of gyration, Biotechnol Prog 19, 544–548.
Young, M. E., Carroad, P. A., and Bell, R. L. (1980) Estimation of Diffusion-Coefficients of Proteins, Biotechnology and Bioengineering 22, 947–955.
Acknowledgments
This investigation was supported by grants from MRC, BBSRC, and Wellcome Trust. We thank Rachel Moore for corrections and comments on the manuscript.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Theveneau, E., Mayor, R. (2011). Beads on the Run: Beads as Alternative Tools for Chemotaxis Assays. In: Wells, C., Parsons, M. (eds) Cell Migration. Methods in Molecular Biology, vol 769. Humana Press. https://doi.org/10.1007/978-1-61779-207-6_30
Download citation
DOI: https://doi.org/10.1007/978-1-61779-207-6_30
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-206-9
Online ISBN: 978-1-61779-207-6
eBook Packages: Springer Protocols