Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1987 Nov 1;105(5):2157–2166. doi: 10.1083/jcb.105.5.2157

Characterization and dynamics of cytoplasmic F-actin in higher plant endosperm cells during interphase, mitosis, and cytokinesis

PMCID: PMC2114876  PMID: 3680376

Abstract

We have identified an F-actin cytoskeletal network that remains throughout interphase, mitosis, and cytokinesis of higher plant endosperm cells. Fluorescent labeling was obtained using actin monoclonal antibodies and/or rhodamine-phalloidin. Video-enhanced microscopy and ultrastructural observations of immunogold-labeled preparations illustrated microfilament-microtubule co-distribution and interactions. Actin was also identified in cell crude extract with Western blotting. During interphase, microfilament and microtubule arrays formed two distinct networks that intermingled. At the onset of mitosis, when microtubules rearranged into the mitotic spindle, microfilaments were redistributed to the cell cortex, while few microfilaments remained in the spindle. During mitosis, the cortical actin network remained as an elastic cage around the mitotic apparatus and was stretched parallel to the spindle axis during poleward movement of chromosomes. This suggested the presence of dynamic cross-links that rearrange when they are submitted to slow and regular mitotic forces. At the poles, the regular network is maintained. After midanaphase, new, short microfilaments invaded the equator when interzonal vesicles were transported along the phragmoplast microtubules. Colchicine did not affect actin distribution, and cytochalasin B or D did not inhibit chromosome transport. Our data on endosperm cells suggested that plant cytoplasmic actin has an important role in the cell cortex integrity and in the structural dynamics of the poorly understood cytoplasm- mitotic spindle interface. F-actin may contribute to the regulatory mechanisms of microtubule-dependent or guided transport of vesicles during mitosis and cytokinesis in higher plant cells.

Full Text

The Full Text of this article is available as a PDF (4.1 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allen N. S., Allen R. D. Cytoplasmic streaming in green plants. Annu Rev Biophys Bioeng. 1978;7:497–526. doi: 10.1146/annurev.bb.07.060178.002433. [DOI] [PubMed] [Google Scholar]
  2. Bajer A. S., Molè-Bajer J. Asters, poles, and transport properties within spindlelike microtubule arrays. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):263–283. doi: 10.1101/sqb.1982.046.01.029. [DOI] [PubMed] [Google Scholar]
  3. Bajer A. S., Molè-Bajer J. Reorganization of microtubules in endosperm cells and cell fragments of the higher plant Haemanthus in vivo. J Cell Biol. 1986 Jan;102(1):263–281. doi: 10.1083/jcb.102.1.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bajer A. S., Sato H., Mole-Bajer J. Video microscopy of colloidal gold particles and immuno-gold labelled microtubules in improved rectified DIC and epi-illumination. Cell Struct Funct. 1986 Sep;11(3):317–330. doi: 10.1247/csf.11.317. [DOI] [PubMed] [Google Scholar]
  5. Barak L. S., Nothnagel E. A., DeMarco E. F., Webb W. W. Differential staining of actin in metaphase spindles with 7-nitrobenz-2-oxa-1,3-diazole-phallacidin and fluorescent DNase: is actin involved in chromosomal movement? Proc Natl Acad Sci U S A. 1981 May;78(5):3034–3038. doi: 10.1073/pnas.78.5.3034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cande W. Z., Lazarides E., McIntosh J. R. A comparison of the distribution of actin and tubulin in the mammalian mitotic spindle as seen by indirect immunofluorescence. J Cell Biol. 1977 Mar;72(3):552–567. doi: 10.1083/jcb.72.3.552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clayton L., Lloyd C. W. Actin organization during the cell cycle in meristematic plant cells. Actin is present in the cytokinetic phragmoplast. Exp Cell Res. 1985 Jan;156(1):231–238. doi: 10.1016/0014-4827(85)90277-0. [DOI] [PubMed] [Google Scholar]
  8. Condeelis J. S. The identification of F actin of the pollen tube and protoplast of Amaryllis belladonna. Exp Cell Res. 1974 Oct;88(2):435–439. doi: 10.1016/0014-4827(74)90269-9. [DOI] [PubMed] [Google Scholar]
  9. De Mey J., Lambert A. M., Bajer A. S., Moeremans M., De Brabander M. Visualization of microtubules in interphase and mitotic plant cells of Haemanthus endosperm with the immuno-gold staining method. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1898–1902. doi: 10.1073/pnas.79.6.1898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Euteneuer U., Schliwa M. Evidence for an involvement of actin in the positioning and motility of centrosomes. J Cell Biol. 1985 Jul;101(1):96–103. doi: 10.1083/jcb.101.1.96. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Forer A. Does actin produce the force that moves a chromosome to the pole during anaphase? Can J Biochem Cell Biol. 1985 Jun;63(6):585–598. doi: 10.1139/o85-077. [DOI] [PubMed] [Google Scholar]
  12. Forer A., Jackson W. T. Actin in spindles of Haemanthus katherinae endosperm. I. General results using various glycerination methods. J Cell Sci. 1979 Jun;37:323–347. doi: 10.1242/jcs.37.1.323. [DOI] [PubMed] [Google Scholar]
  13. Griffith L. M., Pollard T. D. The interaction of actin filaments with microtubules and microtubule-associated proteins. J Biol Chem. 1982 Aug 10;257(15):9143–9151. [PubMed] [Google Scholar]
  14. Gunning B. E., Wick S. M. Preprophase bands, phragmoplasts, and spatial control of cytokinesis. J Cell Sci Suppl. 1985;2:157–179. doi: 10.1242/jcs.1985.supplement_2.9. [DOI] [PubMed] [Google Scholar]
  15. Hartwig J. H., Shevlin P. The architecture of actin filaments and the ultrastructural location of actin-binding protein in the periphery of lung macrophages. J Cell Biol. 1986 Sep;103(3):1007–1020. doi: 10.1083/jcb.103.3.1007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Izant J. G. The role of calcium ions during mitosis. Calcium participates in the anaphase trigger. Chromosoma. 1983;88(1):1–10. doi: 10.1007/BF00329497. [DOI] [PubMed] [Google Scholar]
  17. Kato T., Tonomura Y. Identification of myosin in Nitella flexilis. J Biochem. 1977 Sep;82(3):777–782. doi: 10.1093/oxfordjournals.jbchem.a131754. [DOI] [PubMed] [Google Scholar]
  18. Keith C. H., Ratan R., Maxfield F. R., Bajer A., Shelanski M. L. Local cytoplasmic calcium gradients in living mitotic cells. 1985 Aug 29-Sep 4Nature. 316(6031):848–850. doi: 10.1038/316848a0. [DOI] [PubMed] [Google Scholar]
  19. Kilmartin J. V., Adams A. E. Structural rearrangements of tubulin and actin during the cell cycle of the yeast Saccharomyces. J Cell Biol. 1984 Mar;98(3):922–933. doi: 10.1083/jcb.98.3.922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Langanger G., De Mey J., Adam H. 1,4-Diazobizyklo-[2,2,2]-Oktan (DABCO) verzögert das Ausbleichen von Immunfluoreszenzpräparaten. Mikroskopie. 1983 Sep;40(7-8):237–241. [PubMed] [Google Scholar]
  22. Moeremans M., Daneels G., Van Dijck A., Langanger G., De Mey J. Sensitive visualization of antigen-antibody reactions in dot and blot immune overlay assays with immunogold and immunogold/silver staining. J Immunol Methods. 1984 Nov 30;74(2):353–360. doi: 10.1016/0022-1759(84)90303-x. [DOI] [PubMed] [Google Scholar]
  23. Nothnagel E. A., Barak L. S., Sanger J. W., Webb W. W. Fluorescence studies on modes of cytochalasin B and phallotoxin action on cytoplasmic streaming in Chara. J Cell Biol. 1981 Feb;88(2):364–372. doi: 10.1083/jcb.88.2.364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Palevitz B. A. Actin in the preprophase band of Allium cepa. J Cell Biol. 1987 Jun;104(6):1515–1519. doi: 10.1083/jcb.104.6.1515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pesacreta T. C., Carley W. W., Webb W. W., Parthasarathy M. V. F-actin in conifer roots. Proc Natl Acad Sci U S A. 1982 May;79(9):2898–2901. doi: 10.1073/pnas.79.9.2898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pollard T. D., Cooper J. A. Actin and actin-binding proteins. A critical evaluation of mechanisms and functions. Annu Rev Biochem. 1986;55:987–1035. doi: 10.1146/annurev.bi.55.070186.005011. [DOI] [PubMed] [Google Scholar]
  27. Pollard T. D., Korn E. D. Electron microscopic identification of actin associated with isolated amoeba plasma membranes. J Biol Chem. 1973 Jan 25;248(2):448–450. [PubMed] [Google Scholar]
  28. Pollard T. D., Selden S. C., Maupin P. Interaction of actin filaments with microtubules. J Cell Biol. 1984 Jul;99(1 Pt 2):33s–37s. doi: 10.1083/jcb.99.1.33s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sanger J. M., Sanger J. W. Banding and polarity of actin filaments in interphase and cleaving cells. J Cell Biol. 1980 Aug;86(2):568–575. doi: 10.1083/jcb.86.2.568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sanger J. W. Presence of actin during chromosomal movement. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2451–2455. doi: 10.1073/pnas.72.6.2451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sato M., Leimbach G., Schwarz W. H., Pollard T. D. Mechanical properties of actin. J Biol Chem. 1985 Jul 15;260(14):8585–8592. [PubMed] [Google Scholar]
  32. Schliwa M. Proteins associated with cytoplasmic actin. Cell. 1981 Sep;25(3):587–590. doi: 10.1016/0092-8674(81)90166-5. [DOI] [PubMed] [Google Scholar]
  33. Seagull R. W., Falconer M. M., Weerdenburg C. A. Microfilaments: dynamic arrays in higher plant cells. J Cell Biol. 1987 Apr;104(4):995–1004. doi: 10.1083/jcb.104.4.995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sheetz M. P., Spudich J. A. Movement of myosin-coated fluorescent beads on actin cables in vitro. Nature. 1983 May 5;303(5912):31–35. doi: 10.1038/303031a0. [DOI] [PubMed] [Google Scholar]
  35. Tiwari S. C., Wick S. M., Williamson R. E., Gunning B. E. Cytoskeleton and integration of cellular function in cells of higher plants. J Cell Biol. 1984 Jul;99(1 Pt 2):63s–69s. doi: 10.1083/jcb.99.1.63s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Traas J. A., Doonan J. H., Rawlins D. J., Shaw P. J., Watts J., Lloyd C. W. An actin network is present in the cytoplasm throughout the cell cycle of carrot cells and associates with the dividing nucleus. J Cell Biol. 1987 Jul;105(1):387–395. doi: 10.1083/jcb.105.1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Vantard M., Lambert A. M., De Mey J., Picquot P., Van Eldik L. J. Characterization and immunocytochemical distribution of calmodulin in higher plant endosperm cells: localization in the mitotic apparatus. J Cell Biol. 1985 Aug;101(2):488–499. doi: 10.1083/jcb.101.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Verderame M., Alcorta D., Egnor M., Smith K., Pollack R. Cytoskeletal F-actin patterns quantitated with fluorescein isothiocyanate-phalloidin in normal and transformed cells. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6624–6628. doi: 10.1073/pnas.77.11.6624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wick S. M., Seagull R. W., Osborn M., Weber K., Gunning B. E. Immunofluorescence microscopy of organized microtubule arrays in structurally stabilized meristematic plant cells. J Cell Biol. 1981 Jun;89(3):685–690. doi: 10.1083/jcb.89.3.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wulf E., Deboben A., Bautz F. A., Faulstich H., Wieland T. Fluorescent phallotoxin, a tool for the visualization of cellular actin. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4498–4502. doi: 10.1073/pnas.76.9.4498. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES