Abstract
A method has been developed for the efficient selection of chemotaxis mutants of Dictyostelium discoideum. Mutants defective in the chemotactic response to folate could be enriched up to 30-fold in one round of selection using a chamber in which a compartment that contained the chemoattractant was separated by a sandwich of four nitrocellulose filters from a compartment that contained buffer. Mutagenized cells were placed in the center of the filter layer and exposed to the attractant gradient built up between the compartments for a period of 3-4 h. While wild-type cells moved through the filters in a wave towards the compartment that contained attractant, mutant cells remained in the filter to which they were applied. After several repetitions of the selection procedure, mutants defective in chemotaxis made up 10% of the total cell population retained in that filter. Mutants exhibiting three types of alterations were collected: motility mutants with either reduced speed of movement, or altered rates of turning; a single mutant defective in production of the attractant- degrading enzyme, folate deaminase; and mutants with normal motility but reduced chemotactic responsiveness. One mutant showed drastically reduced sensitivity in folate-induced cGMP production. Morphogenetic alterations of mutants defective in folate chemotaxis are described.
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Selected References
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- 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]
- Barclay S. L., Henderson E. J. Thermosensitive development and tip regulation in a mutant of Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1982 Jan;79(2):505–509. doi: 10.1073/pnas.79.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bernstein R. L., Rossier C., van Driel R., Brunner M., Gerisch G. Folate deaminase and cyclic AMP phosphodiesterase in Dictyostelium discoideum: their regulation by extracellular cyclic AMP and folic acid. Cell Differ. 1981 Mar;10(2):79–86. doi: 10.1016/0045-6039(81)90015-4. [DOI] [PubMed] [Google Scholar]
- Bernstein R. L., Van Driel R. Control of folate deamine activity of Dictyostelium discoideum by cyclic AMP. FEBS Lett. 1980 Oct 6;119(2):249–253. doi: 10.1016/0014-5793(80)80264-x. [DOI] [PubMed] [Google Scholar]
- Bertholdt G., Stadler J., Bozzaro S., Fichtner B., Gerisch G. Carbohydrate and other epitopes of the contact site A glycoprotein of Dictyostelium discoideum as characterized by monoclonal antibodies. Cell Differ. 1985 May;16(3):187–202. doi: 10.1016/0045-6039(85)90516-0. [DOI] [PubMed] [Google Scholar]
- Bonner J. T., Barkley D. S., Hall E. M., Konijn T. M., Mason J. W., O'Keefe G., 3rd, Wolfe P. B. Acrasin, Acrasinase, and the sensitivity to acrasin in Dictyostelium discoideum. Dev Biol. 1969 Jul;20(1):72–87. doi: 10.1016/0012-1606(69)90005-0. [DOI] [PubMed] [Google Scholar]
- Bonner J. T., Hall E. M., Sachsenmaier W., Walker B. K. Evidence for a second chemotactic system in the cellular slime mold, Dictyostelium discoideum. J Bacteriol. 1970 Jun;102(3):682–687. doi: 10.1128/jb.102.3.682-687.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bonner J. T., Hirshfield M. F., Hall E. M. Comparison of a leukocyte and a cellular slime mold chemotaxis test. Exp Cell Res. 1971 Sep;68(1):61–64. doi: 10.1016/0014-4827(71)90586-6. [DOI] [PubMed] [Google Scholar]
- 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]
- Franke J., Kessin R. A defined minimal medium for axenic strains of Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1977 May;74(5):2157–2161. doi: 10.1073/pnas.74.5.2157. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerisch G. Chemotaxis in Dictyostelium. Annu Rev Physiol. 1982;44:535–552. doi: 10.1146/annurev.ph.44.030182.002535. [DOI] [PubMed] [Google Scholar]
- 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]
- Kakebeeke P. I., de Wit R. J., Kohtz S. D., Konijn T. M. Negative chemotaxis in Dictyostelium and Polysphondylium. Exp Cell Res. 1979 Dec;124(2):429–433. doi: 10.1016/0014-4827(79)90218-0. [DOI] [PubMed] [Google Scholar]
- Klein C., Darmon M. Effects of cyclic AMP pulses on adenylate cyclase and the phosphodiesterase inhibitor of D. discoideum. Nature. 1977 Jul 7;268(5615):76–78. doi: 10.1038/268076a0. [DOI] [PubMed] [Google Scholar]
- Klein P., Fontana D., Knox B., Theibert A., Devreotes P. cAMP receptors controlling cell-cell interactions in the development of Dictyostelium. Cold Spring Harb Symp Quant Biol. 1985;50:787–799. doi: 10.1101/sqb.1985.050.01.097. [DOI] [PubMed] [Google Scholar]
- Lo E. K., Coukell M. B., Tsang A. S., Pickering J. L. Physiological and biochemical characterization of aggregation-deficient mutants of Dictyostelium discoideum: detection and response to exogenous cyclic AMP. Can J Microbiol. 1978 Apr;24(4):455–465. doi: 10.1139/m78-075. [DOI] [PubMed] [Google Scholar]
- Matsukuma S., Durston A. J. Chemotactic cell sorting in Dictyostelium discoideum. J Embryol Exp Morphol. 1979 Apr;50:243–251. [PubMed] [Google Scholar]
- McRobbie S. J., Newell P. C. Changes in actin associated with the cytoskeleton following chemotactic stimulation of dictyostelium discoideum. Biochem Biophys Res Commun. 1983 Aug 30;115(1):351–359. doi: 10.1016/0006-291x(83)91011-2. [DOI] [PubMed] [Google Scholar]
- Merkle R. K., Cooper K. K., Rutherford C. L. Localization and levels of cyclic AMP during development of Dictyostelium discoideum. Cell Differ. 1984 Oct;14(4):257–266. doi: 10.1016/0045-6039(84)90014-9. [DOI] [PubMed] [Google Scholar]
- Pan P., Hall E. M., Bonner J. T. Folic acid as second chemotactic substance in the cellular slime moulds. Nat New Biol. 1972 Jun 7;237(75):181–182. doi: 10.1038/newbio237181a0. [DOI] [PubMed] [Google Scholar]
- Pan P., Wurster B. Inactivation of the chemoattractant folic acid by cellular slime molds and identification of the reaction product. J Bacteriol. 1978 Dec;136(3):955–959. doi: 10.1128/jb.136.3.955-959.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rohlf F. J., Davenport D. Simulation of simple models of animal behavior with a digital computer. J Theor Biol. 1969 Jun;23(3):400–424. doi: 10.1016/0022-5193(69)90028-9. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Watts D. J., Ashworth J. M. Growth of myxameobae of the cellular slime mould Dictyostelium discoideum in axenic culture. Biochem J. 1970 Sep;119(2):171–174. doi: 10.1042/bj1190171. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wurster B., Schubiger K. Oscillations and cell development in Dictyostelium discoideum stimulated by folic acid pulses. J Cell Sci. 1977;27:105–114. doi: 10.1242/jcs.27.1.105. [DOI] [PubMed] [Google Scholar]
- van Haastert P. J., De Wit R. J., Konijn T. M. Antagonists of chemoattractants reveal separate receptors for cAMP, folic acid and pterin in Dictyostelium. Exp Cell Res. 1982 Aug;140(2):453–456. doi: 10.1016/0014-4827(82)90139-2. [DOI] [PubMed] [Google Scholar]