Summary
-
1)
The injection of a mushroom drug, Phalloidin (750 μg-1 mg/ml), into the endoplasmic channel of Physarum veins induces an irreversible blockade of the intrinsic contraction-relaxation automaticity of the ectoplasmic tube wall, as measured by tensiometrical methods.
-
2)
The morphological responses to Phalloidin injection include an increase and condensation of cytoplasmic actomyosin sheets bordering the plasmalemma invaginations within the ectoplasmic tube and a more pronounced dense layer of ‘groundplasm’ in the cell cortex. This is in accordance with experiments with other cells as well as with Physarum.
-
3)
The addition of marker particles to the injection solution revealed that the injected substances can be brought into direct contact with the contractile substrate, before newly formed membranes separate off the injection fluid.
-
4)
Since Phalloidin irreversibly transforms oligomeric actin into a filamentous “Phalloidin-actin complex” and because this transformation does not hinder the actin in activating myosin ATPase, it is concluded that the contraction-relaxation cycle of cytoplasmic actomyosin in Physarum involves actin transformations. If these transformations are hindered, e.g. by Phalloidin, one stage and thereby the whole cycle is sustained which results in a blockade of the intrinsic contraction automaticity.
-
5)
The functional importance of actin transformations in the contraction physiology of cytoplasmic actomyosins and cell motility phenomena is discussed.
Similar content being viewed by others
References
Agostini, B., Govindan, V.M., Hofmann, W., Wieland, T.H.: Phalloidin-induced proliferation of actin filaments within rat hepatocytes. Z. Naturforsch. 30c, 793–795 (1975)
Agostini, B., Hofmann, W.: Morphology of Phalloidin intoxication. In: Membrane alterations as basis of liver injury (Popper H., Bianchi L. and Reutter W., eds.), pp. 277–292. Lancester: MTP Press 1977
Bray, D., Thomas, C.: Unpolymerized actin in fibroblasts and brain. J. Mol. Biol. 105, 527–544 (1976)
Camp, W.G.: A method of cultivating myxomycete plasmodia. Bull. Torrey bot. Cl. 63, 205 (1936)
Carlsson, L., Nyström, L.E., Sundkvist, 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)
Daniel, J.W., Rusch, H.P.: The pure culture of Physarum polycephalum on a partially defined soluble medium. J. Gen. Microbiol. 25, 47–59 (1961)
Danker, P., Löw, I., Hasselbach, W., Wieland, T.H.: Interaction of actin with Phalloidin: Polymerization and stabilization of F-actin. Biochim. Biophys. Acta 400, 407–414 (1975)
Dos Remedios, C.G., Barden, J.A.: Activation of myosin ATPase activity by Gd-G-actin. Biochem. Biophys. Res. Commun., Vol. 77, 1339–1346 (1977)
Fleischer, M., Wohlfarth-Bottermann, K.E.: Correlation between tension force generation, fibrillo-genesis and ultrastructure of cytoplasmic actomyosin during isometric and isotonic contractions of protoplasmic strands. Cytobiologie 10, 339–365 (1975)
Gabbiani, G., Montesano, R., Tuchweber, B., Salas, M., Orci, L.: Phalloidin-induced hyperplasia of actin filaments in rat hepatocytes. Lab. Invest. 33, 562–569 (1975)
Goldmann, R., Pollard, T., Rosenbaum, J.: Cell motility. Book A, Vol. 3. Cold Spring Harbor Laboratory 1976
Govindan, V.M., Faulstich, H., Wieland, T.H., Agostini, B., Hasselbach, W.: In vitro effect of Phalloidin on a plasma membrane preparation from rat liver. Naturwissenschaften 59, 521–522 (1972)
Govindan, V.M., Rohr, G., Wieland, T.H., Agostini, B.: Binding of a phallotoxin to protein filaments of plasma membrane of liver cell. Hoppe-Seylers Z. Physiol. Chem. 354, 1159–1161 (1973)
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.W.: Regulation of motility in non-muscle cells. J. Cell Biol. 74, 1–15 (1977)
Hülsmann, N., Wohlfarth-Bottermann, K.E.: Spatio-temporal analysis of contraction dependent surface movements in Physarum polycephalum. Cytobiologie 17, 23–41 (1978a)
Hülsmann, N., Wohlfarth-Bottermann, K.E.: Spatio-temporal relations between protoplasmic streaming and contraction activities in plasmodial veins of Physarum polycephalum. Cytobiologie 17, 317–334 (1978b)
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 the cytoplasmic actomyosin. Cell Tissue Res. 173, 495–528 (1976)
Jahn, T.L., Bovee, E.C.: Protoplasmic movements within cells. Physiol. Rev. 49, 793–861 (1969)
Jahn, W.: Phalloidin hemmt die Bildung eines filamentösen Netzwerkes an der Membran endocytotischer Vakuolen in Leberparenchymzellen. Cytobiologie 15, 452–462 (1977)
Jewell, B.R., Rüegg, J.C.: Oscillatory contraction of insect fibrillar muscle after glycerol extraction. Proc. Royal Soc., B 164, 428–459 (1966)
Kamiya, N.: Protoplasmic streaming. In: Protoplasmatologia 8, 3a (Heilbrunn L.V. and Weber F., eds.), pp. 1–199. Wien: Springer 1959
Komnick, H., Stockem, W., Wohlfarth-Bottermann, K.E.: Cell motility: Mechanism in protoplasmic streaming and ameboid movement. Int. Rev. Cytol. 34, 169–249 (1973)
Korn, E.D.: Biochemistry of actomyosin-dependent cell motility (a review). Proc. Natl. Acad. Sci. U.S.A. 75, 588–599 (1978)
Lengsfeld, A., Jahn, W.: Endocytose an der isolierten perfundierten Rattenleber nach Phalloidinvergiftung. Cytobiologie 9, 391–400 (1974)
Lengsfeld, A., Löw, I., Wieland, T.H., Danker, P., Hasselbach, W.: Interaction of Phalloidin with actin. Proc. Acad. Natl. Sci. U.S.A. 71, 2803–2807 (1974)
Matsumura, F., Hatano, S.: Reversible superprecipitation and bundle formation of plasmodium actomyosin. Biochim. Biophys. Acta 533, 511–523 (1978)
Nagai, R., Kato, L: Cytoplasmic filaments and their assembly into bundles in Physarum plasmodium. Protoplasma 86, 141–158 (1975)
Pollard, T.D., Weihing, R.R.: Actin and myosin and cell movement CRC Crit. Rev. Biochem. 2, 1–65 (1974)
Stockem, W.: Pinocytose und Bewegung von Amöben. Die Reaktion von Amoeba proteus auf verschiedene Markierungssubstanzen. Z. Zellforsch. 74, 372–400 (1966)
Stockem, W.: Die Eignung von Aerosil für die Untersuchung endocytotischer (pinocytotischer) Vorgänge. Mikroskopie 22, 143–147 (1967)
Stockem, W., Weber, K., Wehland, J.: The Influence of microinjected Phalloidin on locomotion, protoplasmic streaming and the organisation of the cytoplasm in Amoeba proteus and Physarum polycephalum. Cytobiologie 18, 114–131 (1978)
Tuchweber, B., Kovacs, K., Khandekar, J.D., Garg, B.D.: Peliosis-like changes induced by Phalloidin in rat liver. J. Med. 4, 327–345 (1973)
Ueda, T., Götz v. Olenhusen, K.: Replacement of endoplasm with artificial media in plasmodial strands of Physarum polycephalum. Effects on contractility and morphology. Exp. Cell Res. 116, 55–62 (1978)
Ueda, T., Götz v. Olenhusen, K., Wohlfarth-Bottermann, K.E.: Reaction of the contractile apparatus in Physarum to injected Ca++, ATP, ADP and 5'AMP. Cytobiologie 18, 76–94 (1978)
Wehland, J., Osborn, M., Weber, K.: Phalloidin-induced actin polymerisation in the cytoplasm of cultured cells interferes with cell locomotion and growth. Proc. Natl. Acad. Sci. U.SA. 74, 5613–5617 (1977)
Wieland, T.: Phallotoxins and microfilaments. In: Molecular basis of motility. (Heilmeyer, L.M.G., Rüegg, J.C. and Wieland, T., eds.), pp. 203–241. Berlin-Heidelberg-New York: Springer 1975
Wieland, T.: Modification of actins by Phallotoxins. Naturwissenschaften 64, 303–309 (1977)
Wieland, T., Wieland, O.: The toxic peptides of Amanita species. In: Microbial toxins (Kadis, S., Ciejler, A. and Ajl, S.J., eds.), pp. 249–280. New York: Academic Press 1972
Wohlfarth-Bottermann, K.E.: Die Kontrastierung tierischer Zellen und Gewebe im Rahmen ihrer elektronenmikroskopischen Untersuchung an ultradünnen Schnitten. Naturwissenschaften 44, 287–288 (1957)
Wohlfarth-Bottermann, K.E.: Weitreichende fibrilläre Protoplasmadifferenzierungen und ihre Bedeutung für die Protoplasmaströmung. II. Lichtmikroskopische Darstellung. Protoplasma 57, 747–761 (1963)
Wohlfarth-Bottermann, K.E.: Weitreichende fibrilläre Protoplasmadifferenzierungen und ihre Bedeutung für die Protoplasmaströmung. III. Entstehung und experimentell induzierbare Musterbildung. Roux' Archiv f. Entwicklungsmech. 156, 371–403 (1965)
Wohlfarth-Bottermann, K.E.: Das Grundplasma. In: Grundlagen der Cytologie (Hirsch, G.C., Ruska, H. und Sitte P., eds.), pp. 123–133. Jena: Fischer 1973
Wohlfarth-Bottermann, K.E.: Plasmalemma invaginations as characteristic constituents of plasmodia of Physarum polycephalum. J. Cell Sci. 16, 23–37 (1974)
Wohlfarth-Bottermann, K.E.: Ursachen von Zellbewegungen. Cytoplasmatische Actomyosine und ihre Bedeutung für Protoplasmaströmungen und Zellmotilität. Leopoldina 21, 85–128 (1975a)
Wohlfarth-Bottermann, K.E.: Weitreichende fibrilläre Protoplasmadifferenzierungen und ihre Bedeutung für die Protoplasmaströmung. X. Die Anordnung der Actomyosin-Fibrillen in experimentell unbeeinflußten Protoplasmaadern von Physarum in situ. Protistologica 11, 19–30 (1975b)
Wohlfarth-Bottermann, K.E.: Tensiometric demonstration of endogenous, oscillating contractions in plasmodia of Physarum polycephalum. Z. Pflanzenphysiol. 76, 14–27 (1975c)
Wohlfarth-Bottermann, K.E.: Oscillating contractions in protoplasmic strands of Physarum: Simultaneous tensiometry of longitudinal and radial rhythms, periodicity analysis and temperature dependence. J. Exp. Biol. 67, 49–59 (1977)
Wohlfarth-Bottermann, K.E., Fleischer, M.: Cycling aggregation patterns of cytoplasmic F-actin coordinated with oscillating tension force generation. Cell Tissue Res. 165, 327–344 (1976)
Wohlfarth-Bottermann, K.E., Götz v. Olenhusen, K.: Oscillating contractions in protoplasmic strands of Physarum: Effects of external Ca++-depletion and Ca++-antagonistic drugs on intrinsic contraction automaticity. Cell Biol. Int. Rpts. 1, 239–247 (1977)
Yoshimoto, Y., Kamiya, N.: Studies on contraction rhythms of the plasmodial strand. II. Role of endoplasmic streaming in synchronization of local rhythms. Protoplasma 95, 111–121 (1978)
Author information
Authors and Affiliations
Additional information
A portion of this work was presented at the Yamada Conference on Cell Motility, VIth International Biophysics Congress, Kyoto, September 1978, Nagoya, Japan
The authors thank Mrs. B. Koeppen for skillful technical assistance and Dr. R.L. Snipes for reading the manuscript. A part of the light microscopical investigations was performed in collaboration with Miss A. Ziesse (Bonn). Dr. U. Lindberg (Uppsala) supplied the purified profilin
Rights and permissions
About this article
Cite this article
von Olenhusen, K.G., Wohlfarth-Bottermann, K.E. Evidence for actin transformation during the contraction-relaxation cycle of cytoplasmic actomyosin: Cycle blockade by Phalloidin injection. Cell Tissue Res. 196, 455–470 (1979). https://doi.org/10.1007/BF00234740
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00234740