Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1993 Oct 2;123(2):345–356. doi: 10.1083/jcb.123.2.345

In vitro models of tail contraction and cytoplasmic streaming in amoeboid cells

PMCID: PMC2119832  PMID: 8408218

Abstract

We have developed a reconstituted gel-sol and contractile model system that mimics the structure and dynamics found at the ectoplasm/endoplasm interface in the tails of many amoeboid cells. We tested the role of gel-sol transformations of the actin-based cytoskeleton in the regulation of contraction and in the generation of endoplasm from ectoplasm. In a model system with fully phosphorylated myosin II, we demonstrated that either decreasing the actin filament length distribution or decreasing the extent of actin filament cross-linking initiated both a weakening of the gel strength and contraction. However, streaming of the solated gel components occurred only under conditions where the length distribution of actin was decreased, causing a self-destruct process of continued solation and contraction of the gel. These results offer significant support that gel strength plays an important role in the regulation of actin/myosin II-based contractions of the tail cortex in many amoeboid cells as defined by the solation-contraction coupling hypothesis (Taylor, D. L., and M. Fechheimer. 1982. Phil. Trans. Soc. Lond. B. 299:185-197). The competing processes of solation and contraction of the gel would appear to be mutually exclusive. However, it is the temporal-spatial balance of the rate and extent of two stages of solation, coupled to contraction, that can explain the conversion of gelled ectoplasm in the tail to a solated endoplasm within the same small volume, generation of a force for the retraction of tails, maintenance of cell polarity, and creation of a positive hydrostatic pressure to push against the newly formed endoplasm. The mechanism of solation-contraction of cortical cytoplasm may be a general component of the normal movement of a variety of amoeboid cells and may also be a component of other contractile events such as cytokinesis.

Full Text

The Full Text of this article is available as a PDF (2.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bray D., White J. G. Cortical flow in animal cells. Science. 1988 Feb 19;239(4842):883–888. doi: 10.1126/science.3277283. [DOI] [PubMed] [Google Scholar]
  2. Brundage R. A., Fogarty K. E., Tuft R. A., Fay F. S. Calcium gradients underlying polarization and chemotaxis of eosinophils. Science. 1991 Nov 1;254(5032):703–706. doi: 10.1126/science.1948048. [DOI] [PubMed] [Google Scholar]
  3. Chen W. T. Mechanism of retraction of the trailing edge during fibroblast movement. J Cell Biol. 1981 Jul;90(1):187–200. doi: 10.1083/jcb.90.1.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Condeelis J. S., Taylor D. L. The contractile basis of amoeboid movement. V. The control of gelation, solation, and contraction in extracts from Dictyostelium discoideum. J Cell Biol. 1977 Sep;74(3):901–927. doi: 10.1083/jcb.74.3.901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Condeelis J. Are all pseudopods created equal? Cell Motil Cytoskeleton. 1992;22(1):1–6. doi: 10.1002/cm.970220102. [DOI] [PubMed] [Google Scholar]
  6. Conrad P. A., Giuliano K. A., Fisher G., Collins K., Matsudaira P. T., Taylor D. L. Relative distribution of actin, myosin I, and myosin II during the wound healing response of fibroblasts. J Cell Biol. 1993 Mar;120(6):1381–1391. doi: 10.1083/jcb.120.6.1381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. De Lozanne A., Spudich J. A. Disruption of the Dictyostelium myosin heavy chain gene by homologous recombination. Science. 1987 May 29;236(4805):1086–1091. doi: 10.1126/science.3576222. [DOI] [PubMed] [Google Scholar]
  8. DeBiasio R. L., Wang L. L., Fisher G. W., Taylor D. L. The dynamic distribution of fluorescent analogues of actin and myosin in protrusions at the leading edge of migrating Swiss 3T3 fibroblasts. J Cell Biol. 1988 Dec;107(6 Pt 2):2631–2645. doi: 10.1083/jcb.107.6.2631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fisher G. W., Conrad P. A., DeBiasio R. L., Taylor D. L. Centripetal transport of cytoplasm, actin, and the cell surface in lamellipodia of fibroblasts. Cell Motil Cytoskeleton. 1988;11(4):235–247. doi: 10.1002/cm.970110403. [DOI] [PubMed] [Google Scholar]
  10. Fukui Y., Lynch T. J., Brzeska H., Korn E. D. Myosin I is located at the leading edges of locomoting Dictyostelium amoebae. Nature. 1989 Sep 28;341(6240):328–331. doi: 10.1038/341328a0. [DOI] [PubMed] [Google Scholar]
  11. Giuliano K. A., Kolega J., DeBiasio R. L., Taylor D. L. Myosin II phosphorylation and the dynamics of stress fibers in serum-deprived and stimulated fibroblasts. Mol Biol Cell. 1992 Sep;3(9):1037–1048. doi: 10.1091/mbc.3.9.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hahn K., DeBiasio R., Taylor D. L. Patterns of elevated free calcium and calmodulin activation in living cells. Nature. 1992 Oct 22;359(6397):736–738. doi: 10.1038/359736a0. [DOI] [PubMed] [Google Scholar]
  13. Hartwig J. H., Stossel T. P. Cytochalasin B and the structure of actin gels. J Mol Biol. 1979 Nov 5;134(3):539–553. doi: 10.1016/0022-2836(79)90366-8. [DOI] [PubMed] [Google Scholar]
  14. Hellewell S. B., Taylor D. L. The contractile basis of ameboid movement. VI. The solation-contraction coupling hypothesis. J Cell Biol. 1979 Dec;83(3):633–648. doi: 10.1083/jcb.83.3.633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hou L., Luby-Phelps K., Lanni F. Brownian motion of inert tracer macromolecules in polymerized and spontaneously bundled mixtures of actin and filamin. J Cell Biol. 1990 May;110(5):1645–1654. doi: 10.1083/jcb.110.5.1645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Höner B., Citi S., Kendrick-Jones J., Jockusch B. M. Modulation of cellular morphology and locomotory activity by antibodies against myosin. J Cell Biol. 1988 Dec;107(6 Pt 1):2181–2189. doi: 10.1083/jcb.107.6.2181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Janmey P. A., Hvidt S., Lamb J., Stossel T. P. Resemblance of actin-binding protein/actin gels to covalently crosslinked networks. Nature. 1990 May 3;345(6270):89–92. doi: 10.1038/345089a0. [DOI] [PubMed] [Google Scholar]
  18. Janson L. W., Kolega J., Taylor D. L. Modulation of contraction by gelation/solation in a reconstituted motile model. J Cell Biol. 1991 Sep;114(5):1005–1015. doi: 10.1083/jcb.114.5.1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Janson L. W., Sellers J. R., Taylor D. L. Actin-binding proteins regulate the work performed by myosin II motors on single actin filaments. Cell Motil Cytoskeleton. 1992;22(4):274–280. doi: 10.1002/cm.970220407. [DOI] [PubMed] [Google Scholar]
  20. Knecht D. A., Loomis W. F. Antisense RNA inactivation of myosin heavy chain gene expression in Dictyostelium discoideum. Science. 1987 May 29;236(4805):1081–1086. doi: 10.1126/science.3576221. [DOI] [PubMed] [Google Scholar]
  21. Kolega J., Janson L. W., Taylor D. L. The role of solation-contraction coupling in regulating stress fiber dynamics in nonmuscle cells. J Cell Biol. 1991 Sep;114(5):993–1003. doi: 10.1083/jcb.114.5.993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Luby-Phelps K., Lanni F., Taylor D. L. The submicroscopic properties of cytoplasm as a determinant of cellular function. Annu Rev Biophys Biophys Chem. 1988;17:369–396. doi: 10.1146/annurev.bb.17.060188.002101. [DOI] [PubMed] [Google Scholar]
  23. Oster G. F., Perelson A. S. The physics of cell motility. J Cell Sci Suppl. 1987;8:35–54. doi: 10.1242/jcs.1987.supplement_8.3. [DOI] [PubMed] [Google Scholar]
  24. Pollard T. D., Cooper J. A. Actin and actin-binding proteins. A critical evaluation of mechanisms and functions. Annu Rev Biochem. 1986;55:987–1035. doi: 10.1146/annurev.bi.55.070186.005011. [DOI] [PubMed] [Google Scholar]
  25. Ris H. The cytoplasmic filament system in critical point-dried whole mounts and plastic-embedded sections. J Cell Biol. 1985 May;100(5):1474–1487. doi: 10.1083/jcb.100.5.1474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sato M., Schwarz W. H., Pollard T. D. Dependence of the mechanical properties of actin/alpha-actinin gels on deformation rate. 1987 Feb 26-Mar 4Nature. 325(6107):828–830. doi: 10.1038/325828a0. [DOI] [PubMed] [Google Scholar]
  27. Simon J. R., Furukawa R. H., Ware B. R., Taylor D. L. The molecular mobility of alpha-actinin and actin in a reconstituted model of gelation. Cell Motil Cytoskeleton. 1988;11(1):64–82. doi: 10.1002/cm.970110107. [DOI] [PubMed] [Google Scholar]
  28. Simon J. R., Taylor D. L. Preparation of a fluorescent analog: acetamidofluoresceinyl-labeled Dictyostelium discoideum alpha-actinin. Methods Enzymol. 1986;134:487–507. doi: 10.1016/0076-6879(86)34115-6. [DOI] [PubMed] [Google Scholar]
  29. Smith S. J. Neuronal cytomechanics: the actin-based motility of growth cones. Science. 1988 Nov 4;242(4879):708–715. doi: 10.1126/science.3055292. [DOI] [PubMed] [Google Scholar]
  30. Stendahl O. I., Stossel T. P. Actin-binding protein amplifies actomyosin contraction, and gelsolin confers calcium control on the direction of contraction. Biochem Biophys Res Commun. 1980 Jan 29;92(2):675–681. doi: 10.1016/0006-291x(80)90386-1. [DOI] [PubMed] [Google Scholar]
  31. Stossel T. P., Chaponnier C., Ezzell R. M., Hartwig J. H., Janmey P. A., Kwiatkowski D. J., Lind S. E., Smith D. B., Southwick F. S., Yin H. L. Nonmuscle actin-binding proteins. Annu Rev Cell Biol. 1985;1:353–402. doi: 10.1146/annurev.cb.01.110185.002033. [DOI] [PubMed] [Google Scholar]
  32. Stossel T. P. The structure of cortical cytoplasm. Philos Trans R Soc Lond B Biol Sci. 1982 Nov 4;299(1095):275–289. doi: 10.1098/rstb.1982.0132. [DOI] [PubMed] [Google Scholar]
  33. Taylor D. L., Blinks J. R., Reynolds G. Contractile basis of ameboid movement. VII. Aequorin luminescence during ameboid movement, endocytosis, and capping. J Cell Biol. 1980 Aug;86(2):599–607. doi: 10.1083/jcb.86.2.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Taylor D. L., Condeelis J. S. Cytoplasmic structure and contractility in amoeboid cells. Int Rev Cytol. 1979;56:57–144. doi: 10.1016/s0074-7696(08)61821-5. [DOI] [PubMed] [Google Scholar]
  35. Taylor D. L., Condeelis J. S., Moore P. L., Allen R. D. The contractile basis of amoeboid movement. I. The chemical control of motility in isolated cytoplasm. J Cell Biol. 1973 Nov;59(2 Pt 1):378–394. doi: 10.1083/jcb.59.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Taylor D. L., Fechheimer M. Cytoplasmic structure and contractility: the solation--contraction coupling hypothesis. Philos Trans R Soc Lond B Biol Sci. 1982 Nov 4;299(1095):185–197. doi: 10.1098/rstb.1982.0125. [DOI] [PubMed] [Google Scholar]
  37. Taylor D. L. The contractile basis of amoeboid movement. IV. The viscoelasticity and contractility of amoeba cytoplasm in vivo. Exp Cell Res. 1977 Mar 15;105(2):413–426. doi: 10.1016/0014-4827(77)90138-0. [DOI] [PubMed] [Google Scholar]
  38. Taylor D. L., Wang Y. L., Heiple J. M. Contractile basis of ameboid movement. VII. The distribution of fluorescently labeled actin in living amebas. J Cell Biol. 1980 Aug;86(2):590–598. doi: 10.1083/jcb.86.2.590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Theriot J. A., Mitchison T. J. Actin microfilament dynamics in locomoting cells. Nature. 1991 Jul 11;352(6331):126–131. doi: 10.1038/352126a0. [DOI] [PubMed] [Google Scholar]
  40. Tilney L. G., Inoué S. Acrosomal reaction of the Thyone sperm. III. The relationship between actin assembly and water influx during the extension of the acrosomal process. J Cell Biol. 1985 Apr;100(4):1273–1283. doi: 10.1083/jcb.100.4.1273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wang Y. L. Exchange of actin subunits at the leading edge of living fibroblasts: possible role of treadmilling. J Cell Biol. 1985 Aug;101(2):597–602. doi: 10.1083/jcb.101.2.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wang Y. L., Lanni F., McNeil P. L., Ware B. R., Taylor D. L. Mobility of cytoplasmic and membrane-associated actin in living cells. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4660–4664. doi: 10.1073/pnas.79.15.4660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wessels D., Soll D. R., Knecht D., Loomis W. F., De Lozanne A., Spudich J. Cell motility and chemotaxis in Dictyostelium amebae lacking myosin heavy chain. Dev Biol. 1988 Jul;128(1):164–177. doi: 10.1016/0012-1606(88)90279-5. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES