Summary
The cytoarchitectural elements ofDictyostelium discoideum amoeba have been visualized by light and electron microscopy in cells prepared with mixtures of glutaraldehyde and Triton-X-100. After negative staining, the peripheral regions of spreading amoebae show a complex meshwork of actin filaments, the majority of which were less than 0.25 microns in length. Multiple branch points, end to side abutments and cross-overs were characteristic features of the actin meshworks. Filopodia extending from the cell periphery consisted of bundles of actin filaments that penetrated into and merged with the actin meshworks in the spreading lamellae. Microtubules emanating from the nucleus associated body penetrated to differing extents into the actin meshworks, sometimes extending close to the cell periphery.Dictyostelium cytoskeletons preparted as described here should prove useful for further studies on the locomotory mechanism.
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References
Berlot CH, Spudich JA, Devreotes PN (1985) Chemoattractant-elicited increase in myosin phosphorylation inDictyostelium. Cell 43: 307–314
Brown SS, Yamamoto K, Spudich JA (1982) A 40,000-dalton protein fromDictyostelium discoideum effects assembly properties of actin in a Ca2+-dependent manner. J Cell Biol 93: 205–210
Cappuccinelli P, Ashworth JM (1976) Effect of microtubule and microfilament function on the cellular slime mouldDictyostelium discoideum. Exp Cell Res 103: 387–393
—,Rubino S, Fighetti M, Unger E (1982) Organization of microtubular system inDictyostelium amoebae. In:Cappuccinelli P, Morris NR (eds) Microtubules in microorganisms. Marcel Dekker, New York and Basel, pp 71–97
Clarke M, Schatten G, Maria D, Spudich JA (1975) Visualization of actin fibers associated with the cell membrane in amoebae ofDictyostelium discoideum. Proc Natl Acad Sci USA 72: 1758–1762
Condeelis J (1979) Isolation of concanavalin A caps during various stages of formation and their association with actin and myosin. J Cell Biol 80: 751–758
— (1981) Microfilament-membrane interactions in cell shape and surface architecture. In:Schweiger HG (ed) International cell biology. Springer, Berlin Heidelberg New York, pp 306–320
—,Vahez M, Carboni JM, De Mey J, Ogihara S (1984) Properties of the 120,000 and 95,000 dalton actin-binding proteins fromDictyostelium discoideum and their possible functions in assembling the cytoplasmic matrix. J Cell Biol 99: (No 1, Pt 2) 119s-126s
Cooke R, Clarke M, von Wedel RJ, Spudich JA (1976) Supramolecular forms ofDictyostelium actin. In:Goldman R, Pollard T, Rosenbaum J (eds) Cell motility. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 575–587
Eckert BS, Lazarides E (1978) Localization of actin inDictyostelium amebas by immunofluorescence. J Cell Biol 77: 714–721
—,Warren RH, Rubin RW (1977) Structural and biochemical aspects of cell motility in amebas ofDictyostelium discoideum. J Cell Biol 72: 339–350
Hellewell SB, Taylor DL (1979) The contractile basis of ameboid movement. J Cell Biol 83: 633–648
Hoglund AS, Karlsson R, Arro E, Fredriksson BE, Lindberg U (1980) Visualization of the peripheral weave of microfilaments in glia cells. J Muscle Res Cell Motil 1: 127–133
Kuczmarski ER, Spudich JA (1930) Regulation of myosin self assembly: phosphorylation ofDictyostelium heavy chain inhibits thick filament formation. Proc Natl Acad Sci USA 77: 7292–7296
Maeda Y, Eguchi G (1977) Polarized structures of cell in the aggregating cellular slime mouldD. discoideum. An electron microscope study. Cell Struct Funct 2: 159–161
Malchow D, Böhme R, Ramsdorf HJ (1981) Regulation of phosphorylation of myosin heavy chain during the chemostatic response ofDictyostelium cells. Eur J Biochem 117: 213–218
Moens PB (1976) Spindle and kinetochore morphology ofDictyostelium discoideum. J Cell Biol 68: 113–122
Pagh KI, Vergara JA, Adelman MR (1985) Improved negative staining of microfilament arrangements in detergent-extractedPhysarum amoeboflagellates. Expl Cell Res 156: 287–293
Roos UP (1982) Morphological and experimental studies on the cytocenter of cellular slime moulds. In:Cappuccinelli P, Morris NR (eds) Microtubules in microorganisms. Marcel Dekker, New York Basel, pp 51–63
—,De Brabander M, Nuydens R (1986) Cell shape and organization of F-actin and microtubules in randomly moving and stationary amoebae ofDictyostelium discoideum. Cell Motility 6: 176–185
Rubino S, Unger E, Fogu G, Cappuccinelli P (1982) Effect of microtubule inhibitors on the tubulin system ofDictyostelium discoideum. Z Allg Mikrobiol 22: 127–131
—,Fighetti M, Unger E, Cappuccinelli P (1983) Location of actin, myosin and microtubular structures during directed locomotion ofDictyostelium amoebae. J Cell Biol 98: 382–390
Salisbury JL, Condeelis JS, Satir P (1983) Receptor-mediated endocytosis: machinery and regulation of the clathrin-coated vesicle pathway. Int Rev Exp Pathol 24: 1–62
Small JV, Celis JE (1978) Filament arrangements in negatively stained cultured cells: the organization of actin. Cytobiologie 16: 308–325
— (1981) Organization of actin in the leading edge of cultured cells: influence of osmium tetroxide and dehydration on the ultrastructure of actin meshworks. J Cell Biol 91: 695–705
- (1982) Organization of actin meshworks in cultured cells: the leading edge. Cold Spring Harbor Symposia on Quantitative Biology, vol XLVI. Cold Spring Harbor Laboratory, pp 599–611
Spudich JA, Spudich A (1982) Cell motility. In:Loomis WF (ed) The development ofDictyostelium discoideum. Academic Press, New York, pp 169–194
Unger E, Rubino S, Weinert T, Cappuccinelli P (1979) Immunofluorescence of the tubulin system in cellular slime moulds. FEMS Microbiol Lett 6: 317–320
Watts DJ, Ashworth JM (1970) Growth of myxamoebae of the cellular slime mouldDictyostelium discoideum in axenic culture. Biochem J 119: 171–174
Yumura S, Mori H, Fukui Y (1984) Localization of actin and myosin for the study of amoeboid movement inDictyostelium using improved immunofluorescence. J Cell Biol 99: 894–899
—,Fukui Y (1985) Reversible cyclic AMP-dependent change in distribution of myosin thick filaments inDictyostelium. Nature 314: 194–196
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Rubino, S., Small, J.V. The cytoskeleton of spreadingDictyostelium amoebae. Protoplasma 136, 63–69 (1987). https://doi.org/10.1007/BF01276319
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DOI: https://doi.org/10.1007/BF01276319