Advertisement
Journal Articles
Mutants lacking myosin II cannot resist forces generated during multicellular morphogenesis
E. Shelden, D.A. Knecht
Journal of Cell Science 1995 108: 1105-1115;

Summary

We have used fluorescent labeling, confocal microscopy and computer-assisted motion analysis to observe and quantify individual wild-type and myosin II mutant cell behavior during early multicellular development in Dictyostelium discoideum. When cultured with an excess of unlabeled wild-type cells, labeled control cells are randomly distributed within aggregation streams, while myosin II mutant cells are found primarily at the lateral edges of streams. Wild-type cells move at average rates of 8.5 +/- 4.9 microns/min within aggregation streams and can exhibit regular periodic movement at 3.5 minute intervals; half as long as the 7 minute period reported previously for isolated cells. Myosin II mutants under the same conditions move at 5.0 +/- 4.8 microns/min, twice as fast as reported previously for isolated myosin II mutant cells, and fail to display regular periodic movement. When removed from aggregation streams myosin II mutant cells move at only 2.5 +/- 2.0 microns/min, while wild-type cells under these conditions move at 5.9 +/- 4.5 microns/min. Analysis of cell morphology further reveals that myosin II mutant cells are grossly and dynamically deformed within wild-type aggregation streams but not when removed from streams and examined in isolation. These data reveal that the loss of myosin II has dramatic consequences for cells undergoing multicellular development. The segregation of mutant cells to aggregation stream edges demonstrates that myosin II mutants are unable to penetrate a multicellular mass of wild-type cells, while the observed distortion of myosin II mutant cells suggests that the cortex of such cells is too flacid to resist forces generated during movement. The increased rate of mutant cell movement and distortion of mutant cell morphology seen within wild-type aggregation streams further argues both that movement of wild-type cells within a multicellular mass can generate traction forces on neighboring cells and that mutant cell morphology and behavior can be altered by these forces. In addition, the distortion of myosin II mutant cells within wild-type aggregation streams indicates that myosin is not required for the formation of cell-cell contacts. Finally, the consequences of the loss of myosin II for cells during multicellular development are much more severe than has been previously revealed for isolated cells. The techniques used here to analyze the behavior of individual cells within multicellular aggregates provide a more sensitive assay of mutant cell phenotype than has been previously available and will be generally applicable to the study of motility and cytoskeletal mutants in Dictyostelium.

REFERENCES

    1. Beug H.,
    2. Katz F. E. and
    3. Gerish G.
    (1973). Dynamics of antigenic membrane sites relating to cell aggregation in Disctyostelium discoideum. J. Cell Biol 56, 647–658
    OpenUrlAbstract/FREE Full Text
    1. Bray D. and
    2. White J. G.
    (1988). Cortical flow in animal cells. Science 239, 883–888
    OpenUrlAbstract/FREE Full Text
    1. Condeelis J.,
    2. Bresnick A.,
    3. Demma M.,
    4. Dharmawardhane S.,
    5. Eddy R.,
    6. Hall A. L.,
    7. Sauterer R. and
    8. Warren V.
    (1990). Mechanisms of amoeboid chemotaxis: an evaluation of the cortical expansion model. Dev. Genet 11, 333–40
    OpenUrlCrossRefPubMedWeb of Science
    1. De Lozanne A. and
    2. Spudich J.
    (1987). Disruption of the Dictyostelium myosin heavy chain gene by homologous recombination. Science 236, 1086–1091
    OpenUrlAbstract/FREE Full Text
    1. Devreotes P.,
    2. Potel M. and
    3. Mackay S.
    (1983). Quantitative analysis of cyclic AMP waves mediating aggregation in Dictyostelium discoideum. Dev. Biol 96, 405–415
    OpenUrlCrossRefPubMedWeb of Science
    1. Devreotes P.
    (1989). Dictyostelium discoideum: a model system for cell-cell interactions in development. Science 245, 1054–1058
    OpenUrlAbstract/FREE Full Text
    1. Eliott S.,
    2. Joss G. H.,
    3. Spudich A. and
    4. Williams K. L.
    (1993). Patterns in Dictyostelium discoideum: the role of myosin II in the transition from the unicellular to the multicellular phase. J. Cell Sci 104, 457–466
    OpenUrlAbstract/FREE Full Text
    1. Fukui Y.,
    2. De Lozanne A. and
    3. Spudich J. A.
    (1990). Structure and function of the cytoskeleton of a Dictyostelium myosin-defective mutant. J. Cell Biol 110, 367–378
    OpenUrlAbstract/FREE Full Text
    1. Futrelle R.,
    2. Traut J. and
    3. McKee W.
    (1982). Cell behavior in Dictyostelium discoideum: preaggregation response to localized cyclic AMP pulses. J.Cell Biol 92, 807–821
    OpenUrlAbstract/FREE Full Text
    1. Harloff C.,
    2. Gerisch G. and
    3. Noegel A. A.
    (1989). Selective elimination of the contact site A protein of Dictyostelium discoideum by gene disruption. Genes Dev 3, 2011–2019
    OpenUrlAbstract/FREE Full Text
    1. Knecht D. A.,
    2. Fuller D. L. and
    3. Loomis W. F.
    (1987). Surface glycoprotein, gp24, involved in early adhesion of Dictyostelium discoideum. Dev. Biol 121, 277–283
    OpenUrlCrossRefPubMed
    1. Kuczmarski E. and
    2. Spudich J.
    (1980). Regulation of myosin self-assembly: phosphorylation of Dictyostelium heavy chain inhibits formation of thick filaments. Proc. Nat. Acad. Sci.USA 77, 7292–7296
    OpenUrlAbstract/FREE Full Text
    1. Lee H.,
    2. Kosciuk M. C.,
    3. Nagele R. G. and
    4. Roisen F. J.
    (1983). Studies on the mechanism of neurulation in the chick: possible involvement of myosin in elevation of neural folds. J. Exp. Zool 225, 449–457
    OpenUrlCrossRefPubMed
    1. Liu G. and
    2. Newell P.
    (1988). Evidence that cyclic GMP regulates myosin interaction with the cytoskeleton during chemotaxis of Dictyostelium. J. Cell Sci 90, 123–129
    OpenUrlAbstract/FREE Full Text
    1. Manstein D.,
    2. Titus M.,
    3. Delozanne A. and
    4. Spudich J.
    (1989). Gene replacement in Dictyostelium- generation of myosin null mutants. EMBO J 8, 923–932
    OpenUrlPubMedWeb of Science
    1. McRobbie S. and
    2. Newell P.
    (1983). Changes in actin associated with the cytoskeleton following chemotactic stimulation of dictyostelium discoideum. Biochem. Biophys. Res. Commun 115, 351–359
    OpenUrlCrossRefPubMedWeb of Science
    1. Morrison A. and
    2. Harwood A.
    (1992). A simple method of generating axenic derivatives of Dictyostelium strains. Exp. Cell Res 199, 383–386
    OpenUrlCrossRefPubMed
    1. Nachmias V. T.,
    2. Fukui Y. and
    3. Spudich J. A.
    (1989). Chemoattractant-elicited translocation of myosin in motile Dictyostelium. Cell Motil. Cytoskel 13, 158–169
    OpenUrlCrossRefPubMedWeb of Science
    1. Newell P. C.,
    2. Europe F. G.,
    3. Liu G.,
    4. Gammon B. and
    5. Wood C. A.
    (1990). Signal transduction for chemotaxis in Dictyostelium amoebae. Semin. Cell Biol 1, 105–113
    OpenUrlPubMed
    1. Pasternak C.,
    2. Spudich J. A. and
    3. Elson E. L.
    (1989). Capping of surface receptors and concomitant cortical tension are generated by conventional myosin. Nature 341, 549–551
    OpenUrlCrossRefPubMed
    1. Peters D.,
    2. Knecht D.,
    3. Loomis W.,
    4. De Lozanne A.,
    5. Spudich J. and
    6. Van Haastert P.
    (1988). Signal transduction, chemotaxis, and cell-aggregation in Dictyostelium discoideum cells without myosin heavy-chain. Dev. Biol 128, 158–163
    OpenUrlCrossRefPubMedWeb of Science
    1. Poot M.,
    2. Kavanagh T. J.,
    3. Kang H. C.,
    4. Haugland R. P. and
    5. Rabinovitch P. S.
    (1991). Flow cytometric analysis of cell cycle-dependent changes in cell thiol level by combining a new laser dye with Hoechst 33342. Cytometry 12, 184–187
    OpenUrlCrossRefPubMedWeb of Science
    1. Sobel J. S.
    (1983). Cell-cell contact modulation of myosin organization in the early mouse embryo. Dev. Biol 100, 207–213
    OpenUrlCrossRefPubMedWeb of Science
    1. Spudich J. A.
    (1989). In pursuit of myosin function. Cell Regul 1, 1–11
    OpenUrlPubMedWeb of Science
    1. Sussman M.
    (1987). Cultivation and synchronous morphogenesis of Dictyostelium under controlled experimental conditions. Meth. Cell Biol 28, 9–29
    OpenUrlPubMedWeb of Science
    1. Traynor D.,
    2. Tasaka M.,
    3. Takeuchi I. and
    4. Williams J.
    (1994). Aberrant pattern formation in myosin heavy chain mutants of Dictyostelium. Development 120, 591–601
    OpenUrlAbstract
    1. Varnum Finney B.,
    2. Edwards K.,
    3. Voss E. and
    4. Soll D.
    (1987). 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. Cytoskel 8, 7–17
    OpenUrlCrossRefPubMedWeb of Science
    1. Wessels D.,
    2. Soll D.,
    3. Knecht D.,
    4. Loomis W.,
    5. Delozanne A. and
    6. Spudich J.
    (1988). Cell motility and chemotaxis in Dictyostelium amebae lacking myosin heavy-chain. Dev. Biol 128, 164–177
    OpenUrlCrossRefPubMedWeb of Science
    1. Wessels D.,
    2. Murray J. and
    3. Soll D. R.
    (1992). Behavior of Dictyostelium amoebae is regulated primarily by the temporal dynamic of the natural cAMP wave. Cell Motil. Cytoskel 23, 145–156
    OpenUrlCrossRefPubMed
    1. Young P. E.,
    2. Richman A. M.,
    3. Ketchum A. S. and
    4. Kiehart D. P.
    (1993). Morphogenesis in Drosophila requires nonmuscle myosin heavy chain function. Genes Dev 7, 29–41
    OpenUrlAbstract/FREE Full Text
    1. Yumura S.,
    2. Mori H. and
    3. Fukui Y.
    (1984). Localization of actin and myosin for the study of ameboid movement in Dictyostelium using improved immunofluorescence. J. Cell Biol 99, 894–899
    OpenUrlAbstract/FREE Full Text
    1. Yumura S. and
    2. Fukui Y.
    (1985). Reversible cyclic AMP-dependent change in distribution of myosin thick filaments in Dictyostelium. Nature 314, 194–196
    OpenUrlCrossRefPubMed
Back to top

 Download PDF

Email
Journal Articles
Mutants lacking myosin II cannot resist forces generated during multicellular morphogenesis
E. Shelden, D.A. Knecht
Journal of Cell Science 1995 108: 1105-1115;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Alerts

Article navigation

Other journals from The Company of Biologists

Advertisement

Follow us on Instagram

Cell science is bursting with beautiful images and over on Instagram, we’re showing them off! Find both JCS and FocalPlane on Instagram for stories and techniques across cell biology.

An interview with Derek Walsh

Professor Derek Walsh is the guest editor of our new special issue Cell Biology of Host-Pathogen Interactions. In an interview, Derek tells us about his work in the field of DNA viruses, the impact of the pandemic on virology and what his role as Guest Editor taught him.

How to improve your scientific writing

"If you are a scientist and you want to succeed, you must become a writer."

How do scientists become master storytellers? We called on our journal Editors, proofreaders and contributors to our community sites for their advice on how to improve your scientific writing.

Meet the preLighters: Jennifer Ann Black

Following the theme of our latest special issue, postdoc Jennifer Ann Black studies replication stress and genome plasticity in Leishmania in Professor Luiz Tosi’s lab in Sao Paolo. We caught up with Jenn (virtually) to hear about her relocation to Brazil mid-pandemic, her research on parasites and what she enjoys about ‘preLighting’.

In our special issue, Chandrakar et al. and Rosazza et al. present their latest work on Leishmania.

Mole – The Corona Files

“There are millions of people around the world who continue to believe that the Terrible Pandemic is a hoax.”

Mole continues to offer his wise words to researchers on how to manage during the COVID-19 pandemic.

JCS and COVID-19

For more information on measures Journal of Cell Science is taking to support the community during the COVID-19 pandemic, please see here.

If you have any questions or concerns, please do not hestiate to contact the Editorial Office.