Abstract
In this chapter, we describe procedures for the microsurgical removal of cells and tissues from early-stage embryos of the amphibian Xenopus laevis. Using simple culture conditions and artificial substrates, these preparations undergo a variety of quantifiable cellular behaviors that closely mimic cell migration in vivo. Two general methods are described. The first includes procedures for obtaining a dorsal marginal zone explant from early gastrulae in order to investigate the sheet-like extension and migration of the mesendoderm that spreads to cover the inner surface of the blastocoel roof in intact embryos. This preparation allows high-resolution analyses of cellular and subcellular events in a contiguous tissue preparation. The second describes methods for the isolation of cranial neural crest cells from tailbud stage embryos. Cranial neural crest tissue cultured in vitro on fibronectin will undergo segmentation and migrate as streams of cells as they do in the developing head. Each of these robust preparations provides an excellent example of the migratory events that are possible to observe in vitro using amphibian embryos.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Keller, R., Davidson, L., Edlund, A., Elul, T., Ezin, M., Shook, D., et al. (2000) Mechanisms of convergence and extension by cell intercalation. Philos. Trans. R Soc. Lond. B Biol. Sci. 355, 897–922.
Keller, R. (2002) Shaping the vertebrate body plan by polarized embryonic cell movements. Science 298, 1950–1954.
Winklbauer, R., Nagel, M., Selchow, A., and Wacker, S. (1996) Mesoderm migration in the Xenopus gastrula. Int. J. Dev. Biol. 40, 305–311.
Sadaghiani, B. and Thiebaud, C. H. (1987) Neural crest development in the Xenopus laevis embryo, studied by interspecific transplantation and scanning electron microscopy. Dev. Biol. 124, 91–110.
Davidson, L. A., Hoffstrom, B. G., Keller, R., and DeSimone, D. W. (2002) Mesendoderm extension and mantle closure in Xenopus laevis gastrulation: combined roles for integrin α5, β1, fibronectin, and tissue geometry. Dev. Biol. 242, 109–129.
Alfandari, D., Cousin, H., Gaultier, A., Hoffstrom, B. G., and DeSimone, D. W. (2003) Integrin α5 β1 supports the migration of Xenopus cranial neural crest on fibronectin. Dev. Biol. 60, 449–464.
Sater, A. K., Steinhardt, R. A., and Keller, R. (1993) Induction of neuronal differentiation by planar signals in Xenopus embryos. Dev. Dyn. 197, 268–280.
Kay, B. K. and Peng, H. B. (1991) Xenopus laevis: Practical Uses in Cell and Molecular Biology, Academic Press, New York, NY.
Sive, H. L., Grainger, R. M., and Harland, R. M. (2000) Early Development of Xenopus Laevis: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Wallingford, J. B., Rowning, B. A., Vogeli, K. M., Rothbacher, U., Fraser, S. E., and Harland, R. M. (2000) Disheveled controls cell polarity during Xenopus gas-trulation. Nature 405, 81–85.
Na, J., Marsden, M., and DeSimone, D. W. (2003) Differential regulation of cell adhesive functions by integrin α subunit cytoplasmic tails in vivo. J. Cell Sci. 116, 2333–2343.
Nieuwkoop, P. D. and Faber, J. (1994) Normal Table of Xenopus laevis (Daudin), Garland Publishing, Inc., New York, NY.
Sokal, R. R. (1981) Biometry, W. H. Freeman and Company, New York, NY.
Ezin, A. M., Skoglund, P., and Keller, R. (2003) The midline (notochord and notoplate) patterns the cell motility underlying convergence and extension of the Xenopus neural plate. Dev. Biol. 256, 100–114.
Davidson, L. A. and Keller, R. E. (1999) Neural tube closure in Xenopus laevis involves medial migration, directed protrusive activity, cell intercalation and convergent extension. Development 126, 4547–4556.
Batschelet, E. (1981) Circular Statistics in Biology, Academic Press, New York, NY.
Elul, T., Koehl, M. A., and Keller, R. (1997) Cellular mechanism underlying neural convergent extension in Xenopus laevis embryos. Dev. Biol. 191, 243–258.
Borchers, A., Epperlein, H. H., and Wedlich, D. (2000) An assay system to study migratory behavior of cranial neural crest cells in Xenopus. Dev. Genes Evol. 210, 217–222.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
DeSimone, D.W., Davidson, L., Marsden, M., Alfandari, D. (2005). The Xenopus Embryo as a Model System for Studies of Cell Migration. In: Guan, JL. (eds) Cell Migration. Methods in Molecular Biology™, vol 294. Humana Press. https://doi.org/10.1385/1-59259-860-9:235
Download citation
DOI: https://doi.org/10.1385/1-59259-860-9:235
Publisher Name: Humana Press
Print ISBN: 978-1-58829-382-4
Online ISBN: 978-1-59259-860-1
eBook Packages: Springer Protocols