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. 1989 Mar 1;108(3):973–984. doi: 10.1083/jcb.108.3.973

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

PMCID: PMC2115405  PMID: 2537839

Abstract

An image processing system was programmed to automatically track and digitize the movement of amebae under phase-contrast microscopy. The amebae moved in a novel chemotaxis chamber designed to provide stable linear attractant gradients in a thin agarose gel. The gradients were established by pumping attractant and buffer solutions through semipermeable hollow fibers embedded in the agarose gel. Gradients were established within 30 min and shown to be stable for at least a further 90 min. By using this system it is possible to collect detailed data on the movement of large numbers of individual amebae in defined attractant gradients. We used the system to study motility and chemotaxis by a score of Dictyostelium discoideum wild-type and mutant strains, including "streamer" mutants which are generally regarded as being altered in chemotaxis. None of the mutants were altered in chemotaxis in the optimal cAMP gradient of 25 nM/mm, with a midpoint of 25 nM. The dependence of chemotaxis on cAMP concentration, gradient steepness, and temporal changes in the gradient were investigated. We also analyzed the relationship between turning behavior and the direction of travel during chemotaxis in stable gradients. The results suggest that during chemotaxis D. discoideum amebae spatially integrate information about local increases in cAMP concentration at various points on the cell surface.

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Selected References

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  1. Alcantara F., Monk M. Signal propagation during aggregation in the slime mould Dictyostelium discoideum. J Gen Microbiol. 1974 Dec;85(2):321–334. doi: 10.1099/00221287-85-2-321. [DOI] [PubMed] [Google Scholar]
  2. BOYDEN S. The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes. J Exp Med. 1962 Mar 1;115:453–466. doi: 10.1084/jem.115.3.453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brenner M., Thoms S. D. Caffeine blocks activation of cyclic AMP synthesis in Dictyostelium discoideum. Dev Biol. 1984 Jan;101(1):136–146. doi: 10.1016/0012-1606(84)90124-6. [DOI] [PubMed] [Google Scholar]
  4. Coukell M. B., Cameron A. M. Genetic locus (stmF) associated with cyclic GMP phosphodiesterase activity in Dictyostelium discoideum maps in linkage group II. J Bacteriol. 1985 Apr;162(1):427–429. doi: 10.1128/jb.162.1.427-429.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Coukell M. B., Cameron A. M., Pitre C. M., Mee J. D. Developmental regulation and properties of the cGMP-specific phosphodiesterase in Dictyostelium discoideum. Dev Biol. 1984 May;103(1):246–257. doi: 10.1016/0012-1606(84)90026-5. [DOI] [PubMed] [Google Scholar]
  6. Darmon M., Brachet P., Da Silva L. H. Chemotactic signals induce cell differentiation in Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3163–3166. doi: 10.1073/pnas.72.8.3163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Deering R. A., Smith M. S., Thompson B. K., Adolf A. C. Gamma-ray-resistant and -sensitive strains of slime mold (Dictyostelium discoideum). Radiat Res. 1970 Sep;43(3):711–728. [PubMed] [Google Scholar]
  8. Devreotes P. N., Steck T. L. Cyclic 3',5' AMP relay in Dictyostelium discoideum. II. Requirements for the initiation and termination of the response. J Cell Biol. 1979 Feb;80(2):300–309. doi: 10.1083/jcb.80.2.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dinauer M. C., Steck T. L., Devreotes P. N. Cyclic 3',5'-AMP relay in Dictyostelium discoideum IV. Recovery of the cAMP signaling response after adaptation to cAMP. J Cell Biol. 1980 Aug;86(2):545–553. doi: 10.1083/jcb.86.2.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dinauer M. C., Steck T. L., Devreotes P. N. Cyclic 3',5'-AMP relay in Dictyostelium discoideum V. Adaptation of the cAMP signaling response during cAMP stimulation. J Cell Biol. 1980 Aug;86(2):554–561. doi: 10.1083/jcb.86.2.554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fisher P. R., Grant W. N., Dohrmann U., Williams K. L. Spontaneous turning behaviour by Dictyostelium discoideum slugs. J Cell Sci. 1983 Jul;62:161–170. doi: 10.1242/jcs.62.1.161. [DOI] [PubMed] [Google Scholar]
  12. Fisher P. R., Smith E., Williams K. L. An extracellular chemical signal controlling phototactic behavior by D. discoideum slugs. Cell. 1981 Mar;23(3):799–807. doi: 10.1016/0092-8674(81)90444-x. [DOI] [PubMed] [Google Scholar]
  13. Futrelle R. P. Dictyostelium chemotactic response to spatial and temporal gradients. Theories of the limits of chemotactic sensitivity and of pseudochemotaxis. J Cell Biochem. 1982;18(2):197–212. doi: 10.1002/jcb.1982.240180207. [DOI] [PubMed] [Google Scholar]
  14. Futrelle R. P., Traut J., McKee W. G. Cell behavior in Dictyostelium discoideum: preaggregation response to localized cyclic AMP pulses. J Cell Biol. 1982 Mar;92(3):807–821. doi: 10.1083/jcb.92.3.807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gerisch G., Fromm H., Huesgen A., Wick U. Control of cell-contact sites by cyclic AMP pulses in differentiating Dictyostelium cells. Nature. 1975 Jun 12;255(5509):547–549. doi: 10.1038/255547a0. [DOI] [PubMed] [Google Scholar]
  16. Gerisch G., Hagmann J., Hirth P., Rossier C., Weinhart U., Westphal M. Early Dictyostelium development: control mechanisms bypassed by sequential mutagenesis. Cold Spring Harb Symp Quant Biol. 1985;50:813–822. doi: 10.1101/sqb.1985.050.01.099. [DOI] [PubMed] [Google Scholar]
  17. Gerisch G., Hülser D., Malchow D., Wick U. Cell communication by periodic cyclic-AMP pulses. Philos Trans R Soc Lond B Biol Sci. 1975 Nov 6;272(915):181–192. doi: 10.1098/rstb.1975.0080. [DOI] [PubMed] [Google Scholar]
  18. Hong C. B., Fontana D. R., Poff K. L. Thermotaxis of Dictyostelium discoideum amoebae and its possible role in pseudoplasmodial thermotaxis. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5646–5649. doi: 10.1073/pnas.80.18.5646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Konijn T. M. Microbiological assay of cyclic 3',5'-AMP. Experientia. 1970 Apr 15;26(4):367–369. doi: 10.1007/BF01896891. [DOI] [PubMed] [Google Scholar]
  20. Konijn T. M., Van De Meene J. G., Bonner J. T., Barkley D. S. The acrasin activity of adenosine-3',5'-cyclic phosphate. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1152–1154. doi: 10.1073/pnas.58.3.1152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lapidus I. R. "Pseudochemotaxis" by micro-organisms in an attractant gradient. J Theor Biol. 1980 Sep 7;86(1):91–103. doi: 10.1016/0022-5193(80)90067-3. [DOI] [PubMed] [Google Scholar]
  22. Mato J. M., Losada A., Nanjundiah V., Konijn T. M. Signal input for a chemotactic response in the cellular slime mold Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4991–4993. doi: 10.1073/pnas.72.12.4991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Meinhardt H., Gierer A. Applications of a theory of biological pattern formation based on lateral inhibition. J Cell Sci. 1974 Jul;15(2):321–346. doi: 10.1242/jcs.15.2.321. [DOI] [PubMed] [Google Scholar]
  24. Potel M. J., Mackay S. A. Preaggregative cell motion in Dictyostelium. J Cell Sci. 1979 Apr;36:281–309. doi: 10.1242/jcs.36.1.281. [DOI] [PubMed] [Google Scholar]
  25. Roos W., Malchow D., Gerisch G. Adenylyl cyclase and the control of cell differentiation in Dictyostelium dicoideum. Cell Differ. 1977 Oct;6(3-4):229–239. doi: 10.1016/0045-6039(77)90018-5. [DOI] [PubMed] [Google Scholar]
  26. Ross F. M., Newell P. C. Streamers: chemotactic mutants of Dictyostelium discoideum with altered cyclic GMP metabolism. J Gen Microbiol. 1981 Dec;127(2):339–350. doi: 10.1099/00221287-127-2-339. [DOI] [PubMed] [Google Scholar]
  27. Segall J. E., Fisher P. R., Gerisch G. Selection of chemotaxis mutants of Dictyostelium discoideum. J Cell Biol. 1987 Jan;104(1):151–161. doi: 10.1083/jcb.104.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Van Haastert P. J., De Wit R. J. Demonstration of receptor heterogeneity and affinity modulation by nonequilibrium binding experiments. The cell surface cAMP receptor of Dictyostelium discoideum. J Biol Chem. 1984 Nov 10;259(21):13321–13328. [PubMed] [Google Scholar]
  29. Van Haastert P. J. Sensory adaptation of Dictyostelium discoideum cells to chemotactic signals. J Cell Biol. 1983 Jun;96(6):1559–1565. doi: 10.1083/jcb.96.6.1559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Varnum-Finney B. J., Voss E., Soll D. R. Frequency and orientation of pseudopod formation of Dictyostelium discoideum amebae chemotaxing in a spatial gradient: further evidence for a temporal mechanism. Cell Motil Cytoskeleton. 1987;8(1):18–26. doi: 10.1002/cm.970080104. [DOI] [PubMed] [Google Scholar]
  31. Varnum-Finney B., Edwards K. B., Voss E., Soll D. R. Amebae 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 ameboid chemotaxis. Cell Motil Cytoskeleton. 1987;8(1):7–17. doi: 10.1002/cm.970080103. [DOI] [PubMed] [Google Scholar]
  32. Varnum B., Edwards K. B., Soll D. R. Dictyostelium amebae alter motility differently in response to increasing versus decreasing temporal gradients of cAMP. J Cell Biol. 1985 Jul;101(1):1–5. doi: 10.1083/jcb.101.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Varnum B., Edwards K. B., Soll D. R. The developmental regulation of single-cell motility in Dictyostelium discoideum. Dev Biol. 1986 Jan;113(1):218–227. doi: 10.1016/0012-1606(86)90124-7. [DOI] [PubMed] [Google Scholar]
  34. Varnum B., Soll D. R. Chemoresponsiveness to cAMP and folic acid during growth, development, and dedifferentiation in Dictyostelium discoideum. Differentiation. 1981;18(3):151–160. doi: 10.1111/j.1432-0436.1981.tb01116.x. [DOI] [PubMed] [Google Scholar]
  35. Varnum B., Soll D. R. Effects of cAMP on single cell motility in Dictyostelium. J Cell Biol. 1984 Sep;99(3):1151–1155. doi: 10.1083/jcb.99.3.1151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Vicker M. G., Schill W., Drescher K. Chemoattraction and chemotaxis in Dictyostelium discoideum: myxamoeba cannot read spatial gradients of cyclic adenosine monophosphate. J Cell Biol. 1984 Jun;98(6):2204–2214. doi: 10.1083/jcb.98.6.2204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Welker D. L., Hirth K. P., Williams K. L. Inheritance of extrachromosomal ribosomal DNA during the asexual life cycle of Dictyostelium discoideum: examination by use of DNA polymorphisms. Mol Cell Biol. 1985 Feb;5(2):273–280. doi: 10.1128/mcb.5.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wilkinson P. C. The measurement of leucocyte chemotaxis. J Immunol Methods. 1982 Jun 11;51(2):133–148. doi: 10.1016/0022-1759(82)90253-8. [DOI] [PubMed] [Google Scholar]
  39. Williams K. L., Newell P. C. A genetic study of aggregation in the cellular slime mould Dictyostelium discoideum using complementation analysis. Genetics. 1976 Feb;82(2):287–307. doi: 10.1093/genetics/82.2.287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Zigmond S. H. Ability of polymorphonuclear leukocytes to orient in gradients of chemotactic factors. J Cell Biol. 1977 Nov;75(2 Pt 1):606–616. doi: 10.1083/jcb.75.2.606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. van Haastert P. J., de Wit R. J., Janssens P. M., Kesbeke F., DeGoede J. G-protein-mediated interconversions of cell-surface cAMP receptors and their involvement in excitation and desensitization of guanylate cyclase in Dictyostelium discoideum. J Biol Chem. 1986 May 25;261(15):6904–6911. [PubMed] [Google Scholar]
  42. van Haastert P. J., van Lookeren Campagne M. M., Ross F. M. Altered cGMP-phosphodiesterase activity in chemotactic mutants of Dictyostelium discoideum. FEBS Lett. 1982 Oct 18;147(2):149–152. doi: 10.1016/0014-5793(82)81029-6. [DOI] [PubMed] [Google Scholar]

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