Skip to main content
SpringerLink
Log in

A contextual framework for characterizing motility and chemotaxis mutants in Dictyostelium discoideum

  • Published:
Journal of Muscle Research & Cell Motility Aims and scope Submit manuscript

Abstract

In the natural aggregation process, Dictyostelium amoebae relay the cAMP signal outwardly through the cell population as symmetric, nondissipating waves. Each cell in turn responds in a specific manner to the different phases of the wave. In the front of each wave, each cell experiences an increasing temporal gradient and positive spatial gradient of cAMP; at the peak of each wave, each cell experiences a cAMP concentration inhibitory to locomotion; and in the back of each wave, each cell experiences a decreasing temporal and negative spatial gradient of cAMP. Protocols are described to analyze the basic motile behavior of mutant cells in the absence of a chemotactic signal, and to test the responsiveness of mutant cells to the individual temporal, spatial and concentration components of a natural wave. The results of such an analysis can then be used to develop realistic models of cell motility and chemotaxis. Examples are described in which this contextual framework has been applied to mutant cell lines. The results of these mutant studies result in a model in which independent parallel regulatory pathways emanating from different phases of the wave effect different phase-specific behaviors.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Alcantara E and Monk M (1974) Signal propagation in the cellular slime mould Dictyostelium discoideum. J Gen Microbiol 85: 321–334.

    Google Scholar 

  • Anjard C, Pinaud R, Kay R and Reymond CD (1992) Overexpression of DdPK2 protein kinase causes rapid development and affects the intra-cellular cAMP pathway of Dictyostelium discoideum. Development 115: 785–790.

    Google Scholar 

  • Aubry L and Firtel RA (1999) Integration of signaling networks that regulate Dictyostelium differentiation. Ann Rev Cell Dev Biol 15: 469–517.

    Google Scholar 

  • Berlot CH, Spudich JA and Devreotes PN (1985) Chemoattractant-elicited increases in myosin phosphorylation in Dictyostelium. Cell 43: 307–314.

    Google Scholar 

  • Berlot CH, Spudich JA and Devreotes PN (1987) Chemoattractant-elicited increases in Dictyostelium myosin phosphorylation are due to changes in myosin localization and increases in kinase activity. J Biol Chem 262: 3918–3926.

    Google Scholar 

  • Bray D (2001) Cell Movements: From Molecules to Motility. Garland Publishing, New York, NY.

    Google Scholar 

  • Chang WT, Thomason PA, Gross JD and Newell PC (1998) Evidence that the RdeA protein is a component of a multistep phosphorelay modulating rate of development in Dictyostelium. EMBO J 17: 2809–2816.

    Google Scholar 

  • Condeelis J (1998) The biochemistry of animal cell crawling. In: Soll DR and Wessels D (eds) Motion Analysis of Living Cells. (pp. 85–100) John Wiley, Inc., Wiley-Liss Division, New York.

    Google Scholar 

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

    Google Scholar 

  • DeLozanne A and Spudich J (1987) Disruption of the Dictyostelium myosin heavy chain gene by homologous recombination. Science 236: 1086–1091.

    Google Scholar 

  • Devreotes P and Zigmond S (1988) Chemotaxis in eukaryotic cells: a focus on leukocytes and Dictyostelium. Ann Rev Cell Biol 4: 649–686.

    Google Scholar 

  • Escalante R, Wessels D, Soll DR and Loomis WF (1997) Chemotaxis to cAMP and slug migration in Dictyostelium both depend on MigA, a BTB protein. Mol Biol Cell 8: 1763–1775.

    Google Scholar 

  • Firtel RA and Chung C (2000) The molecular genetics of chemotaxis: sensing and responding to chemoattractant gradients. Bioessays 22: 603–615.

    Google Scholar 

  • Fisher P, Merkl R and Gerisch G (1989) Quantitative analysis of cell motility and chemotaxis in Dictyostelium discoideum by using an image processing system and a novel chemotaxis chamber providing stationary chemical gradients. J Cell Biol 108: 973–984.

    Google Scholar 

  • Fukui Y, Murray J, Riddelle KS and Soll DR (1991) Cell behavior and actomyosin organization in Dictyostelium during substrate exploration. Cell Struct Funct 16: 289–301.

    Google Scholar 

  • Gerisch G and Keller H (1981) Chemotactic reorientation of granulocytes stimulated with micropipettes containing fMET-Leu-Phe. J Cell Sci 52: 1–10.

    Google Scholar 

  • Hall A, Schlein A and Condeelis J (1988) Relationship of pseudopod extension to chemotactic hormone-induced actin polymerization in ameboid cells. J Cell Biochem 37: 285–299.

    Google Scholar 

  • Hartwell LH, Culotti J, Pringle JR and Reid BJ (1974) Genetic control of the cell division cycle in yeast. Science 183: 46–51.

    Google Scholar 

  • Harwood AJ, Hopper NA, Simon M-N, Bouzid S, Veron M and Williams JG (1992) Multiple roles for cAMP dependent protein kinase during Dictyostelium development. Develop Biol 149: 90–99.

    Google Scholar 

  • Heid P, Voss E and Soll DR (2002) 3D-DIASemb: a computer-assisted system for reconstructing and behaviorally analyzing in 4D every cell and nucleus, and monitoring cytoplasmic flow in a developing embryo. Develop Biol 245: 329–347.

    Google Scholar 

  • Kessin RH (2001) Dictyostelium: Evolution, Cell Biology, and the Development of Multicellularity. Cambridge University Press, Cambridge.

    Google Scholar 

  • Konijn T, van den Meene J, Bonner J and Barkley D (1967) The acrasin activity of adenosine-3′,5′-cyclic phosphate. Proc Natl Acad Sci USA 58: 1152–1154.

    Google Scholar 

  • Kuczmarski ER and Spudich JA (1980) Regulation of myosin self-assembly: phosphorylation of heavy chain inhibits formation of thick filaments. Proc Natl Acad Sci USA 77: 7292–7296.

    Google Scholar 

  • Kuwayama H, Snippe H, Derks M, Roelofs J and Van Haastert P (2001) Identification and characterization of DdPDE3, a cGMP-selective phosphodiesterasae from Dictyostelium. Biochem J 353: 635–644.

    Google Scholar 

  • Laub M and Loomis WF (1998) A molecular network that produces spontaneous oscillations in excitable cells of Dictyostelium. Mol Biol Cell 9: 3521–3532.

    Google Scholar 

  • Lim J, Spiegelman G and Weeks G (2001) RasC is required for optimal activation of adenylyl cyclase and Akt/PKB during aggregation. EMBO J 20: 4490–4499.

    Google Scholar 

  • Loomis WF (1998) Role of PKA in timing of developmental events in Dictyostelium cells. Microbiol Mol Biol Rev 62: 684–694.

    Google Scholar 

  • Loomis WF (1982) The Development of Dictyostelium discoideum. Academic Press, New York.

    Google Scholar 

  • Mann SKO and Firtel RA (1991) A developmentally regulated, putative serine/threonine protein kinase is essential for development in Dictyostelium. Mech Dev 35: 89–101.

    Google Scholar 

  • Mann S, Devreotes P, Eliott S, Jermyn K, Kuspa A, Fechheimer M, Furukawa R, Parent C, Segall J, Shaulsky G, Vardy P, Williams J, Williams K and Firtel RA (1998) Cell Biological, molecular genetic, and biochemical methods used to examine Dictyostelium. In: Cell Biology: A Laboratory Handbook. (pp. 431–465) Academic Press, New York.

    Google Scholar 

  • Mann SKO, Yonemoto WM, Taylor SS and Firtel RA (1992) DdPK3, which plays essential roles during Dictyostelium development, encodes the catalytic subunit of cAMP-dependent protein kinase. Proc Natl Acad Sci USA 89: 10701–10705.

    Google Scholar 

  • Meili R, Ellsworth C, Lee S, Reddy T, Ma H and Firtel RA (1999) Chemoattractant-mediated transient activation and membrane localization of Akt/PKB is required for efficient chemotaxis to cAMP in Dictyostelium. EMBO J 18: 2092–2105.

    Google Scholar 

  • Ostrow BD, Chen P and Chisholm RL (1994) Expression of a myosin regulatory light chain phosphorylation site mutant complements the cytokinesis and developmental defects of Dictyostelium RMLC null cells. J Cell Biol 127: 1945–1955.

    Google Scholar 

  • Pringle JR and Hartwell LH (1981) The Saccharomyces cerevisiae cell cycle. In: Strothern JN, Jones EW and Broach JR (eds) The Molecular Biology of the Yeast Sacchromyces: Life Cycle and Inheritance. (pp. 97–142) Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  • Sanchez-Madrid F and Angel del Pozo M (1999) Leukocyte polarization in cell migration and immune interactions. EMBO J 18: 501–511.

    Google Scholar 

  • Segall J, Fisher P and Gerisch G (1987) Selection of chemotaxis mutants of Dictyostelium discoideum. J Cell Biol 104: 151–161.

    Google Scholar 

  • Shaulsky G, Escalante R and Loomis WF (1996) Developmental signal transduction pathways uncovered by genetic suppressors. Proc Natl Acad Sci USA 93: 15,260–152,665.

    Google Scholar 

  • Shaulsky G, Fuller D and Loomis WF (1998) A cAMP-phosphodiesterase controls PKA-dependent differentiation. Development 125: 691–699.

    Google Scholar 

  • Shutt D, Jenkins LM, Carolan E, Stapleton J, Daniels K, Kennedy R and Soll DR (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.

    Google Scholar 

  • Soll DR (1990) Behavioral studies into the mechanism of eukaryotic chemotaxis. J Chem Ecol 16: 133–150.

    Google Scholar 

  • Soll DR (1995) The use of computers in understanding how animal cells crawl. Int Rev of Cytology 163: 43–104.

    Google Scholar 

  • Soll DR (1999) Computer-assisted three-dimensional reconstruction and motion analysis of living, crawling cells. Comp Med Imag Graph 23: 3–14.

    Google Scholar 

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

  • Soll DR and Wessels D (eds) (1998) Motion Analysis of Living Cells. John Wiley, Inc., Wiley-Liss Division, New York.

    Google Scholar 

  • Soll DR, Voss E, Johnson O and Wessels DJ (2000) Three-dimensional reconstruction and motion analysis of living crawling cells. Scanning 22: 249–257.

    Google Scholar 

  • Soll DR, Wessels D and Sylwester A (1993) The motile behavior of amoebae in the aggregation wave in Dictyostelium discoideum. In: Othmer HG, Maini PK and Murray JD (eds) Experimental and Theoretical Advances in Biological Pattern Formation. (pp. 325–338) NATO ASI Series, Plenium Press, New York.

    Google Scholar 

  • Sussman M (1987) Cultivation and synchronous morphogenesis of Dictyostelium under controlled experimental conditions. Methods Cell Biol 28: 9–30.

    Google Scholar 

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

    Google Scholar 

  • Tomchik KJ and Devreotes PN (1981) Adenosine 3′,5′-monophosphate waves in Dictyostelium discoideum: a demonstration by isotope dilution-fluorography technique. Science 212: 443–446.

    Google Scholar 

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

    Google Scholar 

  • Varnum B and Soll D (1981) Chemoresponsiveness to cAMP and folic acid during growth, development and dedifferentiation in Dictyostelium discoideum. Differentiation 18: 151–160.

    Google Scholar 

  • Varnum B and Soll DR (1984) Effect of cAMP on single cell motility in Dictyostelium. J Cell Biol 99: 1151–1155.

    Google Scholar 

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

    Google Scholar 

  • Varnum-Finney B, Edwards K, Voss E and Soll DR (1987a) Amoebae of Dictyostelium discoideum respond to an increasing temporal gradient of the chemoattractant cAMP with a reduced frequency of turning: evidence for a temporal mechanism in amoeboid chemotaxis. Cell Motil Cytoskel 8: 7–17.

    Google Scholar 

  • Varnum-Finney B, Voss E and Soll DR (1987b) 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.

    Google Scholar 

  • Vawter-Hugart H, Wessels D, Voss E, Minthorn, Murray J and Soll DR (1994) Three dimensional motility cycles exhibited by leukocytes and Dictyostelium discoideum. Cell Motil Cytoskel-Video Suppl 4: 362–364.

    Google Scholar 

  • Ware A, Ware M, Allen F and Lauffenburger D (1999) Shaping up for shipping out: PLC gamma signaling of morphology changes in EGF-stimulated fibroblast migration. Cell Motil Cytoskel 44: 227–233.

    Google Scholar 

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

  • Wessels D, Murray J and Soll DR (1992) Behavior of Dictyostelium amoebae is regulated primarily by the temporal dynamics of the natural cAMP wave. Cell Motil Cytoskel 23: 145–156.

    Google Scholar 

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

    Google Scholar 

  • Wessels D, Schroeder N, Voss E, Hall A, Condeelis J and Soll DR (1989) cAMP mediated inhibition of intracellular particle movement and actin reorganization in Dictyostelium. J Cell Biol 109: 2841–2851.

    Google Scholar 

  • Wessels D, Titus M and Soll DR (1996) A Dictyostelium myosin I plays a crucial role in regulating the frequency of pseudopods formed on the substratum. Cell Motil Cytoskel 33: 64–79.

    Google Scholar 

  • Wessels D, Vawter-Hugart H, Murray J and Soll DR (1994) Three dimensional dynamics of pseudopod formation and the regulation of turning during the motility cycle of Dictyostelium. Cell Motil Cytoskel 27: 1–12.

    Google Scholar 

  • Wessels D, Voss E, Von Bergen N, Burns R, Stites J and Soll DR (1998) A computer-assisted system for reconstructing and interpreting the dynamic three-dimensional relationships of the outer surface, nucleus and pseudopods of crawling cells. Cell Motil Cytoskel 41: 225–246.

    Google Scholar 

  • Wessels D, Zhang H, Reynolds J, Daniels K, Heid P, Liu S, Kuspa A, Shaulsky G, Loomis WF and Soll DR (2000) The internal phosphodiesterase RegA is essential for the suppression of lateral pseudopods during Dictyostelium chemotaxis. Mol Biol Cell 11: 2803–2820.

    Google Scholar 

  • Wilkinson PC (1988) Micropore filter methods for measuring leukocyte chemotaxis. In: Di Sabato G (ed.) Methods Enzymol. (vol. 162, pp. 38–50) Academic Press, New York, NY.

    Google Scholar 

  • Zhang H, Wessels D, Fey P, Daniels K, Chisholm R and Soll DR (2002) Phosphorylation of the myosin regulatory light chain plays a role in cell motility and polarity in Dictyostelium chemotaxis. J Cell Science 115: 1733–1747.

    Google Scholar 

  • Zhang H, Heid PJ, Wessels D, Daniels KJ, Pham T, Loomis WF and Soll DR (2003) Constitutively active protein kinase A disrupts motility and chemotaxis in Dictyostelium discoideum. Eukar Cell 2: 62–75.

    Google Scholar 

  • Zigmond S (1977) The ability of polymorphonuclear leukocytes to orient in gradients of chemotactic factors. J Cell Biol 75: 606–616.

    Google Scholar 

  • Zigmond SH (1978) A new visual assay of leukocyte chemotaxis. In: Gallin JL and Quie PG (eds) Leukocyte Chemotaxis Methods, Physiology and Clinical Implication. (pp. 57–66) Raven Press, New York, NY.

    Google Scholar 

  • Zigmond SH and Hirsch JG (1973) Leukocyte locomotion and chemotaxis. J Exp Med 137: 387–410.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biological Sciences, W.M. Keck Dynamic Image Analysis Facility, University of Iowa, Iowa City, IA, 52242, USA

    David R. Soll, Deborah Wessels, Paul J. Heid & Hui Zhang

Authors
  1. David R. Soll
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Deborah Wessels
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Paul J. Heid
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hui Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Soll, D.R., Wessels, D., Heid, P.J. et al. A contextual framework for characterizing motility and chemotaxis mutants in Dictyostelium discoideum . J Muscle Res Cell Motil 23, 659–672 (2002). https://doi.org/10.1023/A:1024459124427

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1024459124427

Keywords

Search

Navigation