Abstract
In the vegetative state, Dictyostelium amoebae are chemotactic toward pterins released by bacteria, whereas during multicellular development, they become chemotactic to endogenously produced cAMP. A variety of assays have been used to visualize and quantify chemotactic movement. Under-agarose chemotaxis provides a simple and flexible assay that permits high-resolution imaging and quantification of the motility behavior of individual cells and populations by both transmitted light and fluorescence microscopy. The assay requires cells to deform a solid but flexible matrix; therefore, it also provides a way to measure defects in the ability of mutant cells to move in these restrictive conditions.
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Pan, P., Hall, E. M., and Bonner, J. T. (1975) Determination of the active portion of the folic acid molecule in cellular slime mold chemotaxis. J. Bacteriol. 122, 185–191.
Nandini-Kishore, S. G. and Frazier, W. A. (1981) [3H]Methotrexate as a ligand for the folate receptor of Dictyostelium discoideum. Proc. Natl. Acad. Sci. USA 78, 7299–7303.
Alcantara, F. and Monk, M. (1974) Signal propagation during aggregation in the slime mould Dictyostelium discoideum. J. Gen. Microbiol. 85, 321–334.
Devreotes, P. and Janetopoulos, C. (2003) Eukaryotic chemotaxis: distinctions between directional sensing and polarization. J. Biol. Chem. 278, 20,445–20,448.
Rifkin, J. L. (2002) Quantitative analysis of the behavior of Dictyostelium discoideum amoebae: stringency of pteridine reception. Cell Motil. Cytoskeleton 51, 39–48.
Parent, C. A. and Devreotes, P. N. (1996) Molecular genetics of signal transduction in Dictyostelium. Annu. Rev. Biochem. 65, 411–440.
Rifkin, J. L. (2001) Folate reception by vegetative Dictyostelium discoideum amoebae: distribution of receptors and trafficking of ligand. Cell Motil. Cytoskeleton 48, 121–129.
Xiao, Z. and Devreotes, P. N. (1997) Identification of detergent-resistant plasma membrane microdomains in Dictyostelium: enrichment of signal transduction proteins. Mol. Biol. Cell 8, 855–869.
Wessels, D., Murray, J., and Soll, D. R. (1992) Behavior of Dictyostelium amoebae is regulated primarily by the temporal dynamic of the natural cAMP wave. Cell Motil. Cytoskeleton 23, 145–156.
Mato, J. M., Losada, A., Nanjundiah, V., and Konijn, T. M. (1975) Signal input for a chemotactic response in the cellular slime mold Dictyostelium discoideum. Proc. Natl. Acad. Sci. USA 72, 4991–4993.
Ma, L., Janetopoulos, C., Yang, L., Devreotes, P. N., and Iglesias, P. A. (2004) Two complementary, local excitation, global inhibition mechanisms acting in parallel can explain the chemoattractant-induced regulation of PI(3,4,5)P3 response in Dictyostelium cells. Biophys. J. 87, 3764–3774.
Dunn, G. A. and Zicha, D. (1993) Long-term chemotaxis of neutrophils in stable gradients: preliminary evidence of periodic behavior. Blood Cells 19, 25–39.
Shutt, D. C., Jenkins, L. M., Carolan, E. J., et al. (1998) T cell syncytia induced by HIV release. T cell chemoattractants: demonstration with a newly developed single cell chemotaxis chamber. J. Cell Sci. 111(Pt 1), 99–109.
Wilkinson, P. C. (1988) Micropore filter methods for leukocyte chemotaxis. Methods Enzymol. 162, 38–50.
Lauffenburger, D. A., Tranquillo, R. T., and Zigmond, S. H. (1988) Concentration gradients of chemotactic factors in chemotaxis assays. Methods Enzymol. 162, 85–101.
Konijn, T. M. (1970) Microbiological assay of cyclic 3′,5′-AMP. Experientia 26, 367–369.
Bonner, J. T., Kelso, A. P., and Gillmor, R. G. (1966) A new approach to the problem of aggregation in the cellular slime molds. Biol. Bull. 130, 28–42.
Tillinghast, H. S. and Newell, P. C. (1987) Chemotaxis towards pteridines during development of Dictyostelium. J. Cell Sci. 87(Pt 1), 45–53.
Cutler, J. E. and Munoz, J. J. (1974) A simple in vitro method for studies on chemotaxis. Proc. Soc. Exp. Biol. Med. 147, 471–474.
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.
Laevsky, G. and Knecht, D. A. (2001) Under-agarose folate chemotaxis of Dictyostelium discoideum amoebae in permissive and mechanically inhibited conditions. Biotechniques 31, 1140–1149.
Fukui, Y., Yumura, S., Yumura, T. K., and Mori, H. (1986) Agar overlay method: high-resolution immunofluorescence for the study of the contractile apparatus. Methods Enzymol. 134, 573–580.
Neujahr, R., Heizer, C., Albrecht, R., et al. (1997) Three-dimensional patterns and redistribution of myosin II and actin in mitotic Dictyostelium cells. J. Cell Biol. 139, 1793–1804.
Laevsky, G. and Knecht, D. A. (2003) Cross-linking of actin filaments by myosin II is a major contributor to cortical integrity and cell motility in restrictive environments. J. Cell Sci. 116, 3761–3770.
Franke, J. and Kessin, R. (1977) A defined minimal medium for axenic strains of Dictyostelium discoideum. Proc. Natl. Acad. Sci. USA 74, 2157–2161.
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© 2006 Humana Press Inc., Totowa, NJ
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Woznica, D., Knecht, D.A. (2006). Under-Agarose Chemotaxis of Dictyostelium discoideum . In: Eichinger, L., Rivero, F. (eds) Dictyostelium discoideum Protocols. Methods in Molecular Biology™, vol 346. Humana Press. https://doi.org/10.1385/1-59745-144-4:311
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DOI: https://doi.org/10.1385/1-59745-144-4:311
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