Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1994 Aug 1;126(3):713–726. doi: 10.1083/jcb.126.3.713

Lipids trigger changes in the elasticity of the cytoskeleton in plant cells: a cell optical displacement assay for live cell measurements

PMCID: PMC2120140  PMID: 8045935

Abstract

An assay has been developed to quantitatively measure the tension and elasticity of the cytoskeleton in living plant cells. The cell optical displacement assay (CODA) uses a focused laser beam to optically trap and displace transvacuolar and cortical strands through a defined distance within the cell. Results from these experiments provide evidence for the classification of at least two rheologically distinct cytoskeletal assemblies, cortical and transvacuolar, that differ in their tension and response to both signaling molecules and reagents that perturb the cytoskeleton. It is further demonstrated that the tension of the transvacuolar strands can be significantly decreased by the addition of either linoleic acid, 1,2 dioctanoyl-sn-glycerol, or 1,3 dioctanoylglycerol. These decreases in tension could also be induced by lowering the cytoplasmic pH. In contrast, addition of Ca2+, Mg2+, or the ionophore A23187 to the cells caused a considerable increase in the tension of the transvacuolar strands. The data provides evidence that: (a) linoleic acid may be a signaling molecule in plant cells; (b) diacylglycerol functions as a signaling molecule through a protein kinase C-independent pathway mediated by PLA2; and (c) Ca2+ and pH have regulatory roles for controlling cytoskeleton tension and organization.

Full Text

The Full Text of this article is available as a PDF (4.8 MB).

Selected References

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

  1. Ashkin A., Dziedzic J. M. Internal cell manipulation using infrared laser traps. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7914–7918. doi: 10.1073/pnas.86.20.7914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berridge M. J., Irvine R. F. Inositol phosphates and cell signalling. Nature. 1989 Sep 21;341(6239):197–205. doi: 10.1038/341197a0. [DOI] [PubMed] [Google Scholar]
  3. Block S. M. Making light work with optical tweezers. Nature. 1992 Dec 3;360(6403):493–495. doi: 10.1038/360493a0. [DOI] [PubMed] [Google Scholar]
  4. Burridge K. Are stress fibres contractile? Nature. 1981 Dec 24;294(5843):691–692. doi: 10.1038/294691a0. [DOI] [PubMed] [Google Scholar]
  5. Bush D. S., Jones R. L. Measuring intracellular ca levels in plant cells using the fluorescent probes, indo-1 and fura-2 : progress and prospects. Plant Physiol. 1990 Jul;93(3):841–845. doi: 10.1104/pp.93.3.841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cao L. G., Babcock G. G., Rubenstein P. A., Wang Y. L. Effects of profilin and profilactin on actin structure and function in living cells. J Cell Biol. 1992 Jun;117(5):1023–1029. doi: 10.1083/jcb.117.5.1023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Carafoli E. The Ca2+ pump of the plasma membrane. J Biol Chem. 1992 Feb 5;267(4):2115–2118. [PubMed] [Google Scholar]
  8. Clark J. D., Lin L. L., Kriz R. W., Ramesha C. S., Sultzman L. A., Lin A. Y., Milona N., Knopf J. L. A novel arachidonic acid-selective cytosolic PLA2 contains a Ca(2+)-dependent translocation domain with homology to PKC and GAP. Cell. 1991 Jun 14;65(6):1043–1051. doi: 10.1016/0092-8674(91)90556-e. [DOI] [PubMed] [Google Scholar]
  9. Elson E. L. Cellular mechanics as an indicator of cytoskeletal structure and function. Annu Rev Biophys Biophys Chem. 1988;17:397–430. doi: 10.1146/annurev.bb.17.060188.002145. [DOI] [PubMed] [Google Scholar]
  10. Enyedi A., Flura M., Sarkadi B., Gardos G., Carafoli E. The maximal velocity and the calcium affinity of the red cell calcium pump may be regulated independently. J Biol Chem. 1987 May 5;262(13):6425–6430. [PubMed] [Google Scholar]
  11. Evans E. A., La Celle P. L. Intrinsic material properties of the erythrocyte membrane indicated by mechanical analysis of deformation. Blood. 1975 Jan;45(1):29–43. [PubMed] [Google Scholar]
  12. Farmer E. E., Ryan C. A. Interplant communication: airborne methyl jasmonate induces synthesis of proteinase inhibitors in plant leaves. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7713–7716. doi: 10.1073/pnas.87.19.7713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gibbon B. C., Kropf D. L. Cytosolic pH Gradients Associated with Tip Growth. Science. 1994 Mar 11;263(5152):1419–1421. doi: 10.1126/science.263.5152.1419. [DOI] [PubMed] [Google Scholar]
  14. Gong M. C., Fuglsang A., Alessi D., Kobayashi S., Cohen P., Somlyo A. V., Somlyo A. P. Arachidonic acid inhibits myosin light chain phosphatase and sensitizes smooth muscle to calcium. J Biol Chem. 1992 Oct 25;267(30):21492–21498. [PubMed] [Google Scholar]
  15. Grabski S., De Feijter A. W., Schindler M. Endoplasmic Reticulum Forms a Dynamic Continuum for Lipid Diffusion between Contiguous Soybean Root Cells. Plant Cell. 1993 Jan;5(1):25–38. doi: 10.1105/tpc.5.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ha K. S., Exton J. H. Activation of actin polymerization by phosphatidic acid derived from phosphatidylcholine in IIC9 fibroblasts. J Cell Biol. 1993 Dec;123(6 Pt 2):1789–1796. doi: 10.1083/jcb.123.6.1789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hahne G., Hoffmann F. The effect of laser microsurgery on cytoplasmic strands and cytoplasmic streaming in isolated plant protoplasts. Eur J Cell Biol. 1984 Mar;33(2):175–179. [PubMed] [Google Scholar]
  18. Hanahan D. J. Platelet activating factor: a biologically active phosphoglyceride. Annu Rev Biochem. 1986;55:483–509. doi: 10.1146/annurev.bi.55.070186.002411. [DOI] [PubMed] [Google Scholar]
  19. Harper J. F., Sussman M. R., Schaller G. E., Putnam-Evans C., Charbonneau H., Harmon A. C. A calcium-dependent protein kinase with a regulatory domain similar to calmodulin. Science. 1991 May 17;252(5008):951–954. doi: 10.1126/science.1852075. [DOI] [PubMed] [Google Scholar]
  20. Herman B., Pledger W. J. Platelet-derived growth factor-induced alterations in vinculin and actin distribution in BALB/c-3T3 cells. J Cell Biol. 1985 Apr;100(4):1031–1040. doi: 10.1083/jcb.100.4.1031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Irvine R. F. How is the level of free arachidonic acid controlled in mammalian cells? Biochem J. 1982 Apr 15;204(1):3–16. doi: 10.1042/bj2040003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Irvine R. F., Letcher A. J., Dawson R. M. Phosphatidylinositol phosphodiesterase in higher plants. Biochem J. 1980 Oct 15;192(1):279–283. doi: 10.1042/bj1920279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Kolodney M. S., Elson E. L. Correlation of myosin light chain phosphorylation with isometric contraction of fibroblasts. J Biol Chem. 1993 Nov 15;268(32):23850–23855. [PubMed] [Google Scholar]
  25. Kramer R. M., Checani G. C., Deykin D. Stimulation of Ca2+-activated human platelet phospholipase A2 by diacylglycerol. Biochem J. 1987 Dec 15;248(3):779–783. doi: 10.1042/bj2480779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kuo S. C., Sheetz M. P. Force of single kinesin molecules measured with optical tweezers. Science. 1993 Apr 9;260(5105):232–234. doi: 10.1126/science.8469975. [DOI] [PubMed] [Google Scholar]
  27. Leslie C. C., Channon J. Y. Anionic phospholipids stimulate an arachidonoyl-hydrolyzing phospholipase A2 from macrophages and reduce the calcium requirement for activity. Biochim Biophys Acta. 1990 Aug 6;1045(3):261–270. doi: 10.1016/0005-2760(90)90129-l. [DOI] [PubMed] [Google Scholar]
  28. Lloyd C. W. The plant cytoskeleton. Curr Opin Cell Biol. 1989 Feb;1(1):30–35. doi: 10.1016/s0955-0674(89)80033-x. [DOI] [PubMed] [Google Scholar]
  29. Luna E. J., Hitt A. L. Cytoskeleton--plasma membrane interactions. Science. 1992 Nov 6;258(5084):955–964. doi: 10.1126/science.1439807. [DOI] [PubMed] [Google Scholar]
  30. Metcalf T. N., 3rd, Villanueva M. A., Schindler M., Wang J. L. Monoclonal antibodies directed against protoplasts of soybean cells: analysis of the lateral mobility of plasma membrane-bound antibody MVS-1. J Cell Biol. 1986 Apr;102(4):1350–1357. doi: 10.1083/jcb.102.4.1350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Metcalf T. N., 3rd, Wang J. L., Schubert K. R., Schindler M. Lectin receptors on the plasma membrane of soybean cells. Binding and lateral diffusion of lectins. Biochemistry. 1983 Aug 2;22(16):3969–3975. doi: 10.1021/bi00285a037. [DOI] [PubMed] [Google Scholar]
  32. Miller A. J., Vogg G., Sanders D. Cytosolic calcium homeostasis in fungi: roles of plasma membrane transport and intracellular sequestration of calcium. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9348–9352. doi: 10.1073/pnas.87.23.9348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Pace J., Hayman M. J., Galán J. E. Signal transduction and invasion of epithelial cells by S. typhimurium. Cell. 1993 Feb 26;72(4):505–514. doi: 10.1016/0092-8674(93)90070-7. [DOI] [PubMed] [Google Scholar]
  34. Pagano R. E., Longmuir K. J. Phosphorylation, transbilayer movement, and facilitated intracellular transport of diacylglycerol are involved in the uptake of a fluorescent analog of phosphatidic acid by cultured fibroblasts. J Biol Chem. 1985 Feb 10;260(3):1909–1916. [PubMed] [Google Scholar]
  35. Pantoja O., Gelli A., Blumwald E. Voltage-dependent calcium channels in plant vacuoles. Science. 1992 Mar 20;255(5051):1567–1570. doi: 10.1126/science.255.5051.1567. [DOI] [PubMed] [Google Scholar]
  36. Pasternak C., Elson E. L. Lymphocyte mechanical response triggered by cross-linking surface receptors. J Cell Biol. 1985 Mar;100(3):860–872. doi: 10.1083/jcb.100.3.860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Petersen N. O., McConnaughey W. B., Elson E. L. Dependence of locally measured cellular deformability on position on the cell, temperature, and cytochalasin B. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5327–5331. doi: 10.1073/pnas.79.17.5327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Putnam-Evans C. L., Harmon A. C., Cormier M. J. Purification and characterization of a novel calcium-dependent protein kinase from soybean. Biochemistry. 1990 Mar 13;29(10):2488–2495. doi: 10.1021/bi00462a008. [DOI] [PubMed] [Google Scholar]
  39. Qualliotine-Mann D., Agwu D. E., Ellenburg M. D., McCall C. E., McPhail L. C. Phosphatidic acid and diacylglycerol synergize in a cell-free system for activation of NADPH oxidase from human neutrophils. J Biol Chem. 1993 Nov 15;268(32):23843–23849. [PubMed] [Google Scholar]
  40. Rayle D. L., Cleland R. E. The Acid Growth Theory of auxin-induced cell elongation is alive and well. Plant Physiol. 1992 Aug;99(4):1271–1274. doi: 10.1104/pp.99.4.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Ridley A. J., Hall A. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell. 1992 Aug 7;70(3):389–399. doi: 10.1016/0092-8674(92)90163-7. [DOI] [PubMed] [Google Scholar]
  42. Roldan E. R., Fragio C. Diradylglycerols stimulate phospholipase A2 and subsequent exocytosis in ram spermatozoa. Evidence that the effect is not mediated via protein kinase C. Biochem J. 1994 Jan 1;297(Pt 1):225–232. doi: 10.1042/bj2970225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Rosenthal M. D., Lattanzio K. S., Franson R. C. 1,3-Dioctanoylglycerol modulates arachidonate mobilization in human neutrophils and its inhibition by PGBx: evidence of a protein-kinase-C-independent role for diacylglycerols in signal transduction. Biochim Biophys Acta. 1993 May 8;1177(1):79–86. doi: 10.1016/0167-4889(93)90161-h. [DOI] [PubMed] [Google Scholar]
  44. Schaller G. E., Harmon A. C., Sussman M. R. Characterization of a calcium- and lipid-dependent protein kinase associated with the plasma membrane of oat. Biochemistry. 1992 Feb 18;31(6):1721–1727. doi: 10.1021/bi00121a020. [DOI] [PubMed] [Google Scholar]
  45. Scherer G. F., André B. A rapid response to a plant hormone: auxin stimulates phospholipase A2 in vivo and in vitro. Biochem Biophys Res Commun. 1989 Aug 30;163(1):111–117. doi: 10.1016/0006-291x(89)92106-2. [DOI] [PubMed] [Google Scholar]
  46. Schiff P. B., Horwitz S. B. Taxol stabilizes microtubules in mouse fibroblast cells. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1561–1565. doi: 10.1073/pnas.77.3.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Schliwa M., Nakamura T., Porter K. R., Euteneuer U. A tumor promoter induces rapid and coordinated reorganization of actin and vinculin in cultured cells. J Cell Biol. 1984 Sep;99(3):1045–1059. doi: 10.1083/jcb.99.3.1045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Shariff A., Luna E. J. Diacylglycerol-stimulated formation of actin nucleation sites at plasma membranes. Science. 1992 Apr 10;256(5054):245–247. doi: 10.1126/science.1373523. [DOI] [PubMed] [Google Scholar]
  49. 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]
  50. Tsai M. H., Yu C. L., Wei F. S., Stacey D. W. The effect of GTPase activating protein upon ras is inhibited by mitogenically responsive lipids. Science. 1989 Jan 27;243(4890):522–526. doi: 10.1126/science.2536192. [DOI] [PubMed] [Google Scholar]
  51. Wade M. H., de Feijter A. W., Frame M. K. Quantitative fluorescence imaging techniques for the study of organization and signaling mechanisms in cells. Methods Biochem Anal. 1994;37:117–141. doi: 10.1002/9780470110584.ch3. [DOI] [PubMed] [Google Scholar]
  52. Williams D. A., Cody S. H., Gehring C. A., Parish R. W., Harris P. J. Confocal imaging of ionised calcium in living plant cells. Cell Calcium. 1990 Apr;11(4):291–297. doi: 10.1016/0143-4160(90)90006-g. [DOI] [PubMed] [Google Scholar]

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

RESOURCES