Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1983 Mar 1;96(3):857–865. doi: 10.1083/jcb.96.3.857

Filopodelike projections induced with dimethyl sulfoxide and their relevance to cellular polarity in Dictyostelium

PMCID: PMC2112399  PMID: 6682113

Abstract

When 5% dimethyl sulfoxide (DMSO) was applied to Dictyostelium cells, the cells rounded up in shape and cytoplasmic streaming ceased. The cells resumed both cytoplasmic streaming and locomotion in 20 min. SDS PAGE of isolated plasma membrane fractions showed that actin and myosin apparently became dissociated from the plasma membrane by the action of DMSO. Scanning electron microscopy revealed that many filopodelike projections formed on the surface of cells treated with 5% DMSO for 5 min. Interestingly, the projections were formed on a restricted portion of the cell surface. The phagokinetic track technique of Albrecht- Buehler (1977, Cell, 11: 395-404) showed that the projection region corresponded to the anterior part of a migrating cell. The possible relationship between the DMSO-induced projection region on the cell surface and intracellular organization of cell organelles was investigated using serial thin sections. The DMSO-induced projections contained arrays of microfilaments; and the microtubule organizing center (MTOC), nucleus, and vesicular structure were usually located in this order from the anterior end of the cell. The indirect immunofluorescent study using monoclonal anti-alpha-tubulin antibody was performed with a new fixation technique, which greatly improved the phase as well as immunofluorescent microscopy. It was verified that the intracellular positioning of the MTOC and nucleus had significant correlation with the cell polarity. The results show that DMSO is a powerful tool with which to manipulate the cellular microfilaments and to make visible the differentiation in the cortex layer, which apparently is relevant to the intracellular positioning of cell organelles and cell polarity.

Full Text

The Full Text of this article is available as a PDF (4.2 MB).

Selected References

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

  1. Albrecht-Buehler G., Bushnell A. The orientation of centrioles in migrating 3T3 cells. Exp Cell Res. 1979 Apr;120(1):111–118. doi: 10.1016/0014-4827(79)90542-1. [DOI] [PubMed] [Google Scholar]
  2. Albrecht-Buehler G. Filopodia of spreading 3T3 cells. Do they have a substrate-exploring function? J Cell Biol. 1976 May;69(2):275–286. doi: 10.1083/jcb.69.2.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Albrecht-Buehler G. The phagokinetic tracks of 3T3 cells. Cell. 1977 Jun;11(2):395–404. doi: 10.1016/0092-8674(77)90057-5. [DOI] [PubMed] [Google Scholar]
  4. Ames G. F., Nikaido K. Two-dimensional gel electrophoresis of membrane proteins. Biochemistry. 1976 Feb 10;15(3):616–623. doi: 10.1021/bi00648a026. [DOI] [PubMed] [Google Scholar]
  5. Bensadoun A., Weinstein D. Assay of proteins in the presence of interfering materials. Anal Biochem. 1976 Jan;70(1):241–250. doi: 10.1016/s0003-2697(76)80064-4. [DOI] [PubMed] [Google Scholar]
  6. Clarke M., Schatten G., Mazia D., Spudich J. A. Visualization of actin fibers associated with the cell membrane in amoebae of Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1975 May;72(5):1758–1762. doi: 10.1073/pnas.72.5.1758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Coffman D. S., Leichtling B. H., Rickenberg H. V. Phosphoproteins in Dictyostelium discoideum. J Supramol Struct Cell Biochem. 1981;15(4):369–385. doi: 10.1002/jsscb.1981.380150407. [DOI] [PubMed] [Google Scholar]
  8. Condeelis J. Isolation of concanavalin A caps during various stages of formation and their association with actin and myosin. J Cell Biol. 1979 Mar;80(3):751–758. doi: 10.1083/jcb.80.3.751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. De Robertis E. M., Longthorne R. F., Gurdon J. B. Intracellular migration of nuclear proteins in Xenopus oocytes. Nature. 1978 Mar 16;272(5650):254–256. doi: 10.1038/272254a0. [DOI] [PubMed] [Google Scholar]
  10. Eckert B. S., Lazarides E. Localization of actin in Dictyostelium amebas by immunofluorescence. J Cell Biol. 1978 Jun;77(3):714–721. doi: 10.1083/jcb.77.3.714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Eckert B. S., Warren R. H., Rubin R. W. Structural and biochemical aspects of cell motility in amebas of Dictyostelium discoideum. J Cell Biol. 1977 Feb;72(2):339–350. doi: 10.1083/jcb.72.2.339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Filosa M. F., Fukui Y. Dimethyl sulfoxide inhibits capping of surface receptors. Cell Biol Int Rep. 1981 Jun;5(6):575–579. doi: 10.1016/s0309-1651(81)80008-2. [DOI] [PubMed] [Google Scholar]
  13. Fowler V., Taylor D. L. Spectrin plus band 4.1 cross-link actin. Regulation by micromolar calcium. J Cell Biol. 1980 May;85(2):361–376. doi: 10.1083/jcb.85.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fukui Y. Formation of multinuclear cells induced by dimethyl sulfoxide: inhibition of cytokinesis and occurrence of novel nuclear division in Dictyostelium cells. J Cell Biol. 1980 Jul;86(1):181–189. doi: 10.1083/jcb.86.1.181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fukui Y. Intranuclear actin bundles induced by dimethyl sulfoxide in interphase nucleus of Dictyostelium. J Cell Biol. 1978 Jan;76(1):146–157. doi: 10.1083/jcb.76.1.146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fukui Y., Katsumaru H. Dynamics of nuclear actin bundle induction by dimethyl sulfoxide and factors affecting its development. J Cell Biol. 1980 Jan;84(1):131–140. doi: 10.1083/jcb.84.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fukui Y., Katsumaru H. Nuclear actin bundles in Amoeba, Dictyostelium and human HeLa cells induced by dimethyl sulfoxide. Exp Cell Res. 1979 May;120(2):451–455. doi: 10.1016/0014-4827(79)90412-9. [DOI] [PubMed] [Google Scholar]
  18. Jacobson B. S. Actin binding to the cytoplasmic surface of the plasma membrane isolated from Dictyostelium discoideum. Biochem Biophys Res Commun. 1980 Dec 31;97(4):1493–1498. doi: 10.1016/s0006-291x(80)80034-9. [DOI] [PubMed] [Google Scholar]
  19. Kuriyama R., Sato C., Fukui Y., Nishibayashi S. In vitro nucleation of microtubules from microtubule-organizing center prepared from cellular slime mold. Cell Motil. 1982;2(3):257–272. doi: 10.1002/cm.970020306. [DOI] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. Luna E. J., Fowler V. M., Swanson J., Branton D., Taylor D. L. A membrane cytoskeleton from Dictyostelium discoideum. I. Identification and partial characterization of an actin-binding activity. J Cell Biol. 1981 Feb;88(2):396–409. doi: 10.1083/jcb.88.2.396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Malech H. L., Root R. K., Gallin J. I. Structural analysis of human neutrophil migration. Centriole, microtubule, and microfilament orientation and function during chemotaxis. J Cell Biol. 1977 Dec;75(3):666–693. doi: 10.1083/jcb.75.3.666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mooseker M. S. Brush border motility. Microvillar contraction in triton-treated brush borders isolated from intestinal epithelium. J Cell Biol. 1976 Nov;71(2):417–433. doi: 10.1083/jcb.71.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  25. Osborn M., Weber K. Dimethylsulfoxide and the ionophore A23187 affect the arrangement of actin and induce nuclear actin paracrystals in PtK2 cells. Exp Cell Res. 1980 Sep;129(1):103–114. doi: 10.1016/0014-4827(80)90335-3. [DOI] [PubMed] [Google Scholar]
  26. RODRIGUEZ J., DEINHARDT F. Preparation of a semipermanent mounting medium for fluorescent antibody studies. Virology. 1960 Oct;12:316–317. doi: 10.1016/0042-6822(60)90205-1. [DOI] [PubMed] [Google Scholar]
  27. Sanger J. W., Sanger J. M., Kreis T. E., Jockusch B. M. Reversible translocation of cytoplasmic actin into the nucleus caused by dimethyl sulfoxide. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5268–5272. doi: 10.1073/pnas.77.9.5268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Spudich J. A. Biochemical and structural studies of actomyosin-like proteins from non-muscle cells. II. Purification, properties, and membrane association of actin from amoebae of Dictyostelium discoideum. J Biol Chem. 1974 Sep 25;249(18):6013–6020. [PubMed] [Google Scholar]
  29. Taylor D. L., Condeelis J. S. Cytoplasmic structure and contractility in amoeboid cells. Int Rev Cytol. 1979;56:57–144. doi: 10.1016/s0074-7696(08)61821-5. [DOI] [PubMed] [Google Scholar]

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

RESOURCES