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Planta

, Volume 156, Issue 5, pp 466–474 | Cite as

Wound-healing motility in the green alga Ernodesmis: calcium ions and metabolic energy are required

  • John W. La ClaireII
Article

Abstract

Wounding a giant cell of the marine alga Ernodesmis verticillata (Kützing) Børgesen (Chlorophyta) induces two concomitant motility phenomena: longitudinal contraction of the protoplasm away from the wound site, and centripetal contraction of the cut end around the central vacuole. Healing is complete within 30 min of wounding. Mechanical extrusion of the protoplasm into the medium with a teasing needle is followed by contraction of the protoplasm into numerous spherical protoplasts within 60 min. Utilizing a simple defined medium, it is shown that motility is almost completely inhibited by the absence of exogenous free Ca2+, with 5.0 mM ethylene glycol bis-(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid present. This inhibition is reversible by rinsing the cells with Ca2+-containing medium. Similarly, extruded cytoplasm fails to exhibit motility in Ca2+-free medium. The threshold concentration of exogenous free Ca2+ is approx. 10-7 M for wound-induced contraction. The ions Ba2+, Cd2+ and Sr2+ will substitute for Ca2+, but the rate of contraction is one-half that with Ca2+ present. Although darkness has no inhibitory effect, lower temperature (15°C), cyanide, or micromolar amounts of phosphorylation uncouplers reversibly slow protoplasmic motility in wounded cells and extruded cytoplasm. Carbonylcyanide m-chlorophenylhydrazone and carbonylcyanide p-trifluoromethoxyphenylhydrazone are especially potent inhibitors. These results indicate that cellular wound healing utilizes metabolic energy and requires exogenous free Ca2+, implying that motility in Ernodesmis is a true contractile process. Since 1.0 mM La3+ completely and reversibly prevents contraction, it is postulated that Ca2+ fluxes may actually trigger motility.

Key words

Calcium and motility/wound healing Cell repair Chlorophyta Ernodesmis Motility Wound healing 

Abbreviations

CCCP

carbonylcyanide m-chlorophenylhydrazone

DMSO

dimethylsulfoxide

DNP

2,4-dinitrophenol

EGTA

ethylene glycol bis-(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid

FCCP

carbonylcyanide p-trifluoromethoxyphenylhydrazone

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References

  1. Allen, N.S., Allen, R.D. (1978) Cytoplasmic streaming in green plants. Annu. Rev. Biophys. Bioeng 7, 497–526Google Scholar
  2. Allen, R.D. (1981) Motility. J. Cell Biol. 91, 148s-155sGoogle Scholar
  3. Bluemink, J.G. (1972) Cortical wound healing in the amphibian egg: an electron microscopical study. J. Ultrastruct. Res. 41, 95–114Google Scholar
  4. Borle, A.B. (1978) On the difficulty of assessing the role of extracellular calcium in cell function. Ann. N. Y. Acad. Sci. 307, 431–432Google Scholar
  5. Borle, A.B. (1981) Control, modulation, and regulation of cell calcium. Rev. Physiol. Biochem. Pharmacol. 90, 13–153Google Scholar
  6. Burr, F.A., Evert, R.F. (1972) A cytochemical study of the wound-healing protein in Bryopsis hypnoides. Cytobios 6, 199–215Google Scholar
  7. Burr, F.A., West, J.A. (1971) Protein bodies in Bryopsis hypnoides: their relationship to wound-healing and branch septum development. J. Ultrastruct. Res. 35, 476–498Google Scholar
  8. Carafoli, E., Crompton, M. (1978) The regulation of intracellular calcium. Curr. Top. Membr. Transp. 10, 151–216Google Scholar
  9. Dillon, L.S. (1981) Ultrastructure, macromolecules, and evolution. Plenum Press, New YorkGoogle Scholar
  10. dos Remedios, C.G. (1981) Lanthanide ion probes of calcium-binding sites on cellular membranes. Cell Calcium 2, 29–51Google Scholar
  11. Gillet, C., Lefebvre, J. (1980) Calcium binding in the cell wall of Nitella. In: Plant membrane transport: current conceptual issues, pp. 421–422, Spanswick, R.M., Lucas, W.J., Dainty, J., eds. Elsevier/North-Holland Biomedical Press, AmsterdamGoogle Scholar
  12. Gingell, D. (1970) Contractile responses at the surface of an amphibian egg. J. Embryol. Exp. Morphol. 23, 583–609Google Scholar
  13. Good, N., Izawa, S. (1966) Uncoupling and energy transfer inhibition in photophosphorylation. Curr. Top. Bioenerg. 1, 75–112Google Scholar
  14. Häder, D.-P. (1982) Coupling of photomovement and photosynthesis in desmids. Cell Motility 2, 73–82Google Scholar
  15. Higashi-Fujime, S. (1980) Active movement in vitro of bundles of microfilaments isolated from Nitella cell. J. Cell Biol. 87, 569–578Google Scholar
  16. Hinnen, R., Miyamoto, H., Racker, E. (1979) Ca2+ translocation in Ehrlich ascites tumor cells. J. Membr. Biol. 49, 309–324Google Scholar
  17. Ishizawa, K., Enomoto, S., Wada, S. (1979) Germination and photo-induction of polarity in the spherical cells regenerated from protoplasm fragments of Boergesenia forbesii. Bot. Mag. Tokyo 92, 173–186Google Scholar
  18. Jagendorf, A.T. (1975) Mechanism of photophosphorylation. In: Bioenergetics of photosynthesis, pp. 413–492, Govindjee, ed. Academic Press, New YorkGoogle Scholar
  19. Jeon, K.W., Jeon, M.S. (1975) Cytoplasmic filaments and cellular wound healing in Amoeba proteus. J. Cell Biol. 67, 243–249Google Scholar
  20. Kamiya, N. (1981) Physical and chemical basis of cytoplasmic streaming. Annu. Rev. Plant Physiol. 32, 205–236Google Scholar
  21. Kato, T., Tonomura, Y. (1977) Identification of myosin in Nitella flexilis. J. Biochem. 82, 777–782Google Scholar
  22. Kessler, H. (1980) On the selective adsorption of cations in the cell wall of the green alga Valonia utricularis. Helgol. Wiss. Meeresunters. 34, 151–158Google Scholar
  23. La Claire, J.W., II (1982) Cytomorphological aspects of wound healing in selected Siphonocladales (Chlorophyceae). J. Phycol. 18, 379–384Google Scholar
  24. Lehninger, A.L. (1975) Biochemistry, 2nd edn. Worth Publications, New YorkGoogle Scholar
  25. Lien, S., San Pietro, A. (1981) Effect of uncouplers on anaerobic adaptation of hydrogenase activity in C. reinhardtii. Biochem. Biophys. Res. Comm. 103, 139–147Google Scholar
  26. Luckenbill, L.M. (1971) Dense material associated with wound closure in the axolotl egg (A. mexicanum). Exp. Cell Res. 66, 263–267Google Scholar
  27. Luft, J.H. (1971) Ruthenium red and violet. I. Chemistry, purification, methods of use for electron microscopy and mechanism of action. Anat. Rec. 171, 347–368Google Scholar
  28. Martin, R.B., Richardson, F.S. (1979) Lanthanides as probes for calcium in biological systems. Q. Rev. Biophys. 12, 181–209Google Scholar
  29. Ohsuka, K., Inoue, A. (1979) Identification of myosin in a flowering plant, Egeria densa. J. Biochem. 85, 375–378Google Scholar
  30. Park, B.H. (1981) Cell motility. In: Advanced cell biology, pp. 195–225, Schwartz, L.M., Azar, M.M., eds. Van Nostrand Reinhold Co., New YorkGoogle Scholar
  31. Reed, K.C., Bygrave, F.L. (1975) Methodology for in vitro studies of Ca2+ transport. Anal Biochem. 67, 44–54Google Scholar
  32. Schmid, R.W., Reilley, C.N. (1957) New complexon for titration of calcium in the presence of magnesium. Anal Chem. 29, 264–268Google Scholar
  33. Seitz, K. (1979) Cytoplasmic streaming and cyclosis of chloroplasts. In: Encyclopedia of plant physiology, N.S., vol. 7: Physiology of movements, pp. 150–169, Haupt, W., Feinleib, M.E., eds. Springer, Berlin Heidelberg New YorkGoogle Scholar
  34. Stassart, J.M., Neirinckx, L., Dajaegere, R. (1981) The interactions between monovalent cations and calcium during their adsorption on isolated cell walls and absorption by intact barley roots. Ann. Bot. 47, 647–652Google Scholar
  35. Stebbings, H., Hyams, J.S. (1979) Cell motility. Longman, New YorkGoogle Scholar
  36. Sverdrup, H.U., Johnson, M.W., Fleming, R.H. (1942) The oceans. Their physics, chemistry, and general biology. Prentice-Hall, New YorkGoogle Scholar
  37. Szubinska, B. (1971) “New membrane” formation in Amoeba proteus upon injury of individual cells. Electron microscope observations. J. Cell Biol. 49, 747–772Google Scholar
  38. Szubinska, B. (1978) Closure of the plasma membrane around microneedle in Amoeba proteus. An ultrastructural study. Exp. Cell Res. 111, 105–115Google Scholar
  39. Tatewaki, M., Nagata, K. (1970) Surviving protoplasts in vitro and their development in Bryopsis. J. Phycol. 6, 101–103Google Scholar
  40. Tendel, J., Haupt, W. (1981) Mechanische und energetische Grundlagen der lichtabhängigen Gestaltänderung des Mougeotia-Chloroplasten. Z. Pflanzenphysiol. 104, 169–185Google Scholar
  41. Tominaga, Y., Tazawa, M. (1981) Reversible inhibition of cytoplasmic streaming by intracellular Ca2+ in tonoplast-free cells of Chara australis. Protoplasma 109, 103–111Google Scholar
  42. Vahey, M., Scordilis, S.P. (1980) Contractile proteins from the tomato. Can. J. Bot. 58, 797–801Google Scholar
  43. Wagner, G. (1979) Actomyosin as a basic mechanism of movement in animals and plants. In: Encyclopedia of plant physiology, N.S., vol. 7: Physiology of movements, pp. 114–126, Haupt, W., Feinleib, M.E., eds. Springer, Berlin Heidelberg New YorkGoogle Scholar
  44. Weiss, G.B. (1974) Cellular pharmacology of lanthanum. Annu. Rev. Pharmacol. 14, 343–354Google Scholar
  45. Williamson, R.E. (1975) Cytoplasmic streaming in Chara: a cell model activated by ATP and inhibited by cytochalasin B. J. Cell Sci. 17, 655–668Google Scholar
  46. Williamson, R.E. (1979) Filaments associated with the endoplasmic reticulum in the streaming cytoplasm of Chara corallina. Eur. J. Cell Biol. 20, 177–183Google Scholar
  47. Williamson, R.E., Ashley, C.C. (1982) Free Ca2+ and cytoplasmic streaming in the alga Chara. Nature (London) 296, 647–651Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • John W. La ClaireII
    • 1
  1. 1.Department of BotanyUniversity of TexasAustinUSA

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