Summary
Microinjection of DNAase I, which is known to form a specific complex with G-actin, induces characteristic changes in cytoplasmic streaming, locomotion and morphology of the contractile apparatus of A. proteus. Light microscopical studies show pronounced streaming originating from the uroid and/or the retracting pseudopods, which ceases 10–15 min after injection of DNAase I, at a time when ultrastructural studies show that the actin filament system is very much reduced. These results suggest that a controlled reversible equilibrium between soluble and polymerized forms of actin is a necessary requirement for amoeboid movement.
The topographic distribution of contractile filaments beneath the plasma membrane visualized by correlated light- and electron microscopy of DNAase I-injected cells establishes the importance of the membrane-bound filamentous layer for three major aspects of streaming: (1) Streaming originates by local contractions of a cell membrane-associated filament layer at the uroid and/or retracting pseudopods, creating a pressure flow. (2) This flow continues beneath the membrane, which is stabilized by filaments in the lateral regions between the posterior end, with a high hydrostatic pressure, and the anterior end, with a low hydrostatic pressure. (3) Pseudopods or extending areas are created by a local destabilization of the cell periphery caused by the separation of the filamentous layer from the plasma membrane.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Carlsson, L., Nyström, L.-E., Sunkvist, I., Markey, F., Lindberg, U.: Actin polymerizability is influenced by profilin, a low molecular weight protein in non-muscle cells. J. Mol. Biol. 115, 465–483 (1977)
Comly, L.T.: Microfilaments in Chaos carolinensis: membrane association, distribution and heavy meromyosin binding in the glycerinated cell. J. Cell Biol. 58, 230–237 (1973)
D'Haese, J., Komnick, H.: Fine structure and contraction of isolated muscle actomyosin. I. Evidence for a sliding mechanism by means of oligomeric myosin. Z. Zellforsch. 134, 411–426 (1972a)
D'Haese, J., Komnick, H.: Fine structure and contraction of isolated muscle actomyosin. II. Formation of myosin filaments and their effect on contraction. Z. Zellforsch. 134, 427–434 (1972b)
Fujinami, N., Kageyama, T.: Circus movement in dissociated embryonic cells of a teleost, Oryzias latipes. J. Cell Sci. 19, 169–181 (1975)
Grebecki, A., Grebecka, L.: Morphodynamic types of Amoeba proteus: a terminological proposal. Protistologica 14, 349–358 (1978)
Gruenstein, E., Rich, A., Weihing, R.R.: Actin associated with plasma membranes from 3T3 and HeLa cells. J. Cell Biol. 64, 223–234 (1975)
Haberey, M.: Räumliche Anordnung von Plasmafilamenten bei Thecamoeba sphaeronucleolus. Cytobiologie 8, 61–75 (1973)
Haberey, M., Stockem, W.: Amoeba proteus: Morphologie, Zucht und Verhalten. Mikrokosmos 60, 33–42 (1971)
Harris, H.E., Weeds, A.G.: Platelet actin: sub-cellular distribution and association with profilin. FEBS Letters 90, 84–88 (1978)
Hauser, M.: Demonstration of membrane-associated and oriented microfilaments in Amoeba proteus by means of a Schiff base/glutaraldehyde fixative. Cytobiologie 18, 95–106 (1978)
Hinssen, H.: Actin in isoliertem Grundplasma von Physarum polycephalum. Cytobiologie 5, 146–164 (1972)
Hinssen, H., D'Haese, J.: Synthetic fibrils from Physarum actomyosin — self assembly, organization and contraction. Cytobiologie 13, 132–157 (1976)
Hitchcock, S.E., Carlsson, L., Lindberg, U.: DNAase I-induced depolymerization of actin filaments. In: Cell Motility, R. Goldman, T. Pollard, and J. Rosenbaum (eds.), pp. 545–559. Cold Spring Harbor Laboratory, 1976
Holtfreter, J.: Structure, motility and locomotion in isolated embryonic cells. J. Morphol. 79, 27–62 (1946)
Isenberg, G., Wohlfarth-Bottermann, K.E.: Transformation of cytoplasmic actin. Importance for the organization of the contractile gel reticulum and the contraction-relaxation cycle of cytoplasmic actomyosin. Cell Tissue Res. 173, 495–528 (1976)
Jahn, W.: Phalloidin hemmt die Bildung eines filamentösen Netzwerkes an der Membran endocytotischer Vakuolen in Leberparenchymzellen. Cytobiologie 15, 452–462 (1977)
Kamiya, N.: The motive force of endoplasmic streaming in the amoeba. In: Primitive Motile Systems in Cell Biology, R.D. Allen and N. Kamiya (eds.), pp. 257–278. New York: Academic Press 1964
Klein, H.P., Stockem, W.: Pinocytosis and locomotion of amoeba. XII. Dynamics of cell membrane during induced pinocytosis in A. proteus. Cell Tissue Res., 197, 263–279 (1979)
Korn, E.D.: Biochemistry of actomyosin-dependent cell motility. Proc. Natl. Acad. Sci. U.S.A. 75, 588–599 (1978)
Korn, E.D., Wright, P.L.: Macromolecular composition of an amoeba plasma membrane. J. Biol. Chem. 248, 439–447 (1973)
Korohoda, W., Stockem, W.: Experimentally induced destabilization of the cell membrane and cell surface activity in Amoeba proteus. Cytobiologie 12, 93–110 (1975a)
Korohoda, W., Stockem, W.: On the nature of hyaline zones in the cytoplasm of Amoeba proteus. Microsc. Acta 77, 129–141 (1975b)
Lazarides, E., Lindberg, U.: Actin is the naturally occurring inhibitor of deoxyribonuclease I. Proc. Natl. Acad. Sci. U.S.A. 71, 4742–4746 (1974)
Lindberg, U., Carlsson, L., Nyström, L.-E., Kannan, K.K., Cid-Dresdner, H., Lövgren, S., Jörnvall, H.: Crystallization of a non-muscle actin. In: Cell Motility, R. Goldman, T. Pollard, and J. Rosenbaum (eds.), pp. 561–573, Cold Spring Harbor Laboratory, 1976
Markey, F., Lindberg, U., Eriksson, L.: Human platelets contain profilin, a potential regulator of actin polymerizability. FEBS Letters 88, 75–79 (1978)
Mast, S.O.: Structure, movement, locomotion and stimulation in amoebae. J. Morphol. 41, 347–425 (1926)
Pollard, T.D.: Functional implications of the biochemical and structural properties of cytoplasmic contractile proteins. In: Molecules and Cell Movement, S. Inoué and R.E. Stevens (eds.), pp. 259–286. New York: Raven Press, 1975
Pollard, T.D., Korn, E.D.: Electron microscopic identification of actin associated with isolated amoeba plasma membranes. J. Biol. Chem. 248, 448–450 (1973)
Rinaldi, R.A., Baker, W.R.: A sliding filament model of amoeboid motion. J. Theor. Biol. 23, 463–474 (1969)
Schroeder, T.E.: Dynamics of the contractile ring. In: Molecules and Cell Movement, S. Inoué and R.E. Stevens (eds.), pp. 305–334. New York: Raven Press 1975
Spurr, A.R.: A low-viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res. 26, 31–43 (1969)
Stockem, W., Weber, K., Wehland, J.: The influence of microinjected phalloidin on locomotion, protoplasmic streaming and cytoplasmic organization in Amoeba proteus and Physarum polycephalum. Cytobiologie 18, 114–131 (1978)
Taylor, D.L.: Motile model systems of amoeboid movement. In: Cell Motility, R. Goldman, T. Pollard, and J. Rosenbaum (eds.), pp. 797–821. Cold Spring Harbor Laboratory 1976
Taylor, D.L.: The contractile basis of amoeboid movement. IV. The visco-elasticity and contractility of amoeba cytoplasm in vivo. Exp. Cell Res. 105, 413–426 (1977)
Taylor, D.L., Rhodes, J.A., Hammond, S.A.: The contractile basis of amoeboid movement. II. Structure and contractility of motile extracts and plasmalemma-ectoplasm ghosts. J. Cell Biol. 70, 123–143 (1976)
Tilney, L.G.: The role of actin in nonmuscle cell motility. In: Molecules and Cell Movement, S. Inoué and R.E. Stevens (eds.), pp. 339–388. New York: Raven Press 1975
Tilney, L.G., Mooseker, M.: Actin in the brush border of epithelial cells of the chicken intestine. Proc. Natl. Acad. Sci. U.S.A. 68, 2611–2615 (1971)
Weber, K., Groeschel-Stewart, U.: Antibody to myosin: the specific visualization of myosin-containing filaments in non-muscle cells. Proc. Natl. Acad. Sci. U.S.A. 71, 4561–4564 (1974)
Wehland, J., Osborn, M., Weber, K.: Phalloidin-induced actin polymerization in the cytoplasm of cultured cells interferes with cell locomotion and growth. Proc. Natl. Acad. Sci. U.S.A. 74, 5613–5617 (1977)
Wehland, J., Stockem, W., Weber, K.: Cytoplasmic streaming in Amoeba proteus is inhibited by the actin specific drug phalloidin. Exp. Cell Res. 115, 451–454 (1978)
Wessels, N.K., Spooner, B.S., Luduena, M.: Surface movements, microfilaments and cell locomotion. In: Locomotion of Tissue Cells, K. Porter and D.W. Fitzsimons (eds.), pp. 53–82. Amsterdam-London-New York: Elsevier, Excerpta Medica. North Holland, Assoc. Scientific Publ., 1973
Wieland, T.: Modification of actins by phallotoxins. Naturwissenschaften 64, 303–309 (1977)
Author information
Authors and Affiliations
Additional information
W. Gawlitta was supported by a grant (No. 21619) from the Konrad-Adenauer-Stiftung
The authors wish to thank Dr. Mary Osborn for reading the manuscript
Rights and permissions
About this article
Cite this article
Wehland, J., Weber, K., Gawlitta, W. et al. Effects of the actin-binding protein DNAase I on cytoplasmic streaming and ultrastructure of Amoeba proteus . Cell Tissue Res. 199, 353–372 (1979). https://doi.org/10.1007/BF00236075
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00236075