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Analyzing Chemotaxis Using Dictyostelium discoideum as a Model System

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Cell Migration in Inflammation and Immunity

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 239))

Abstract

Dictyostelium discoideum are simple eukaryotic amoebas that when starved of a food source have the remarkable ability to spontaneously aggregate and differentiate to form a cluster of spores on top of a stalk of vacuolated cells (see Fig. 1 and ref. 1). The process of aggregation, although quite fascinating to simply observe, has proven extremely powerful for the molecular dissection of chemotaxis signaling pathways. Mechanisms of chemoattractant sensing discovered in D. discoideum have been tested in higher eukaryotic systems and were found to be strikingly similar (2). In particular, a paradigm for directional sensing has been derived from studies in D. discoideum (3).

Dictyostelium discoideum development. Development on development buffer (see Subheading 3.2.1. ) agar at 0 h (A), 7 h (B), 24 h (C), and 24 h, 10 times higher magnification (D) after plating. Wild-type plaque (E) and aggregation-deficient plaque (F) on Sm/KA plates after 5 d.

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References

  1. Parent, C. A. and Devreotes, P. N. (1996) Molecular genetics of signal transduction in Dictyostelium. Annu. Rev. Biochem. 65, 411–440.

    Article  PubMed  CAS  Google Scholar 

  2. Devreotes, P. N. and Zigmond, S. H. (1988) Chemotaxis in eukaryotic cells: a focus on leukocytes and Dictyostelium, Annu. Rev. Cell Biol. 4, 649–686.

    Article  PubMed  CAS  Google Scholar 

  3. Parent, C. A. and Devreotes, P. N. (1999) A cell’s sense of direction. Science 284, 765–770.

    Article  PubMed  CAS  Google Scholar 

  4. Hadwiger, J. A., Lee, S., and Firtel, R. A. (1994) The G alpha subunit G alpha 4 couples to pterin receptors and identifies a signaling pathway that is essential for multicellular development in Dictyostelium, Proc. Natl. Acad. Sci. USA 91, 10,566–10,570.

    Article  PubMed  CAS  Google Scholar 

  5. Xiao, Z., Zhang, N., Murphy, D. B., and Devreotes, P. N. (1997) Dynamic distribution of chemoattractant receptors in living cells during chemotaxis and persistent stimulation. J. Cell Biol. 139, 365–374.

    Article  PubMed  CAS  Google Scholar 

  6. Jin, T., Zhang, N., Long, Y., Parent, C. A., and Devreotes, P. N. (2000) Localization of the G protein betagamma complex in living cells during chemotaxis. Science 287, 1034–1036.

    Article  PubMed  CAS  Google Scholar 

  7. Lilly, P. J. and Devreotes, P. N. (1995) Chemoattractant and GTP gamma S-mediated stimulation of adenylyl cyclase in Dictyostelium requires translocation of CRAC to membranes. J. Cell Biol. 129, 1659–1665.

    Article  PubMed  CAS  Google Scholar 

  8. Parent, C. A., Blacklock, B. J., Froehlich, W. M., Murphy, D. B., and Devreotes, P. N. (1998) G Protein signaling events are activated at the leading edge of chemotactic cells. Cell 95, 81–91.

    Article  PubMed  CAS  Google Scholar 

  9. Iijima, M. and Devreotes, P. (2002) Tumor suppressor PTEN mediates sensing of chemoattractant gradients. Cell 109, 599–610.

    Article  PubMed  CAS  Google Scholar 

  10. Funamoto, S., Meili, R., Lee, S., Parry, L., and Firtel, R. A. (2002) Spatial and temporal regulation of 3-phosphoinositides by PI 3-kinase and PTEN mediates chemotaxis. Cell 109, 611–623.

    Article  PubMed  CAS  Google Scholar 

  11. Rickert, P., Weiner, O. D., Wang, F., Bourne, H. R., and Servant, G. (2000) Leukocytes navigate by compass: roles of PI3Kgamma and its lipid products. Trends Cell Biol. 10, 466–473.

    Article  PubMed  CAS  Google Scholar 

  12. Wang, F., Herzmark, P., Weiner, O. D., Srinivasan, S., Servant, G., and Bourne, H. R. (2002) Lipid products of PI(3)Ks maintain persistent cell polarity and directed motility in neutrophils. Nat. Cell Biol. 4, 513–518.

    Article  PubMed  CAS  Google Scholar 

  13. Weiner, O. D., Neilsen, P. O., Prestwich, G. D., Kirschner, M. W., Cantley, L. C., and Bourne, H. R. (2002) A PtdInsP(3)-and Rho GTPase-mediated positive feedback loop regulates neutrophil polarity. Nat. Cell Biol. 4, 509–513.

    Article  PubMed  CAS  Google Scholar 

  14. Kuspa, A. and Loomis, W. F. (1992) Tagging developmental genes in Dictyostelium by restriction enzyme-mediated integration of plasmid DNA. Proc. Natl. Acad. Sci. USA 89, 8803–8807.

    Article  PubMed  CAS  Google Scholar 

  15. Robinson, D. N. and Spudich, J. A. (2000) Dynacortin, a genetic link between equatorial contractility and global shape control discovered by library complementation of a Dictyostelium discoideum cytokinesis mutant. J. Cell Biol. 150, 823–838.

    Article  PubMed  CAS  Google Scholar 

  16. van Es, S. and Devreotes, P. N. (1999) Molecular basis of localized responses during chemotaxis in amoebae and leukocytes. Cell. Mol. Life Sci. 55, 1341–1351.

    Article  PubMed  Google Scholar 

  17. Sutoh, K. (1993) A transformation vector for Dictyostelium discoideum with a new selectable marker bsr. Plasmid 30, 150–154.

    Article  PubMed  CAS  Google Scholar 

  18. Kuwayama, H., Obara, S., Morio, T., Katoh, M., Urushihara, H., and Tanaka, Y. (2002) PCR-mediated generation of a gene disruption construct without the use of DNA ligase and plasmid vectors. Nucleic Acids Res. 30, E2.

    Article  PubMed  Google Scholar 

  19. Blaauw, M., Linskens, M. H., and van Haastert, P. J. (2000) Efficient control of gene expression by a tetracycline-dependent transactivator in single Dictyostelium discoideum cells. Gene 252, 71–82.

    Article  PubMed  CAS  Google Scholar 

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© 2004 Humana Press Inc., Totowa, NJ

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Landree, M.A., Devreotes, P.N. (2004). Analyzing Chemotaxis Using Dictyostelium discoideum as a Model System. In: D’Ambrosio, D., Sinigaglia, F. (eds) Cell Migration in Inflammation and Immunity. Methods in Molecular Biology™, vol 239. Humana Press. https://doi.org/10.1385/1-59259-435-2:91

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  • DOI: https://doi.org/10.1385/1-59259-435-2:91

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-102-8

  • Online ISBN: 978-1-59259-435-1

  • eBook Packages: Springer Protocols

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