Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1993 Jun 2;121(6):1369–1383. doi: 10.1083/jcb.121.6.1369

Cytoskeletal remodeling during growth cone-target interactions

PMCID: PMC2119712  PMID: 8509456

Abstract

Reorganization of the cytoskeleton of neuronal growth cones in response to environmental cues underlies the process of axonal guidance. Most previous studies addressing cytoskeletal changes during growth cone pathfinding have focused on the dynamics of a single cytoskeletal component. We report here an investigation of homophilic growth cone- target interactions between Aplysia bag cell neurons using digitally enhanced video microscopy, which addresses dynamic interactions between actin filaments and microtubules. After physical contact of a growth cone with a physiological target, mechanical coupling occurred after a delay; and then the growth cone exerted forces on and displaced the target object. Subsequent to coupling, F-actin accumulation was observed at the target contact zone, followed by preferential microtubule extension to the same site. After successful target interactions, growth cones typically moved off highly adhesive poly-L- lysine substrates into native target cell surfaces. These events were associated with modulation of both the direction and rate of neurite outgrowth: growth cone migration was typically reoriented to a trajectory along the target interaction axis and rates of advance increased by about one order of magnitude. Directed microtubule movements toward the contact site appeared to be F-actin dependent as target site-specific microtubule extension and bundling could be reversibly randomized by micromolar levels of cytochalasin B in a characteristic manner. Our results suggest that target contacts can induce focal F-actin assembly and reorganization which, in turn, guides target site-directed microtubule redistribution.

Full Text

The Full Text of this article is available as a PDF (5.1 MB).

Selected References

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

  1. Abercrombie M., Heaysman J. E., Pegrum S. M. The locomotion of fibroblasts in culture. 3. Movements of particles on the dorsal surface of the leading lamella. Exp Cell Res. 1970 Oct;62(2):389–398. doi: 10.1016/0014-4827(70)90570-7. [DOI] [PubMed] [Google Scholar]
  2. Bentley D., Toroian-Raymond A. Disoriented pathfinding by pioneer neurone growth cones deprived of filopodia by cytochalasin treatment. Nature. 1986 Oct 23;323(6090):712–715. doi: 10.1038/323712a0. [DOI] [PubMed] [Google Scholar]
  3. Bray D. Axonal growth in response to experimentally applied mechanical tension. Dev Biol. 1984 Apr;102(2):379–389. doi: 10.1016/0012-1606(84)90202-1. [DOI] [PubMed] [Google Scholar]
  4. Bray D. Mechanical tension produced by nerve cells in tissue culture. J Cell Sci. 1979 Jun;37:391–410. doi: 10.1242/jcs.37.1.391. [DOI] [PubMed] [Google Scholar]
  5. Bray D. Surface movements during the growth of single explanted neurons. Proc Natl Acad Sci U S A. 1970 Apr;65(4):905–910. doi: 10.1073/pnas.65.4.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bridgman P. C., Dailey M. E. The organization of myosin and actin in rapid frozen nerve growth cones. J Cell Biol. 1989 Jan;108(1):95–109. doi: 10.1083/jcb.108.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bridgman P. C., Kachar B., Reese T. S. The structure of cytoplasm in directly frozen cultured cells. II. Cytoplasmic domains associated with organelle movements. J Cell Biol. 1986 Apr;102(4):1510–1521. doi: 10.1083/jcb.102.4.1510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cheney R. E., Mooseker M. S. Unconventional myosins. Curr Opin Cell Biol. 1992 Feb;4(1):27–35. doi: 10.1016/0955-0674(92)90055-h. [DOI] [PubMed] [Google Scholar]
  9. Cooper M. S., Schliwa M. Electrical and ionic controls of tissue cell locomotion in DC electric fields. J Neurosci Res. 1985;13(1-2):223–244. doi: 10.1002/jnr.490130116. [DOI] [PubMed] [Google Scholar]
  10. Correas I., Padilla R., Avila J. The tubulin-binding sequence of brain microtubule-associated proteins, tau and MAP-2, is also involved in actin binding. Biochem J. 1990 Jul 1;269(1):61–64. doi: 10.1042/bj2690061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Danowski B. A. Fibroblast contractility and actin organization are stimulated by microtubule inhibitors. J Cell Sci. 1989 Jun;93(Pt 2):255–266. doi: 10.1242/jcs.93.2.255. [DOI] [PubMed] [Google Scholar]
  12. Dennerll T. J., Joshi H. C., Steel V. L., Buxbaum R. E., Heidemann S. R. Tension and compression in the cytoskeleton of PC-12 neurites. II: Quantitative measurements. J Cell Biol. 1988 Aug;107(2):665–674. doi: 10.1083/jcb.107.2.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dennerll T. J., Lamoureux P., Buxbaum R. E., Heidemann S. R. The cytomechanics of axonal elongation and retraction. J Cell Biol. 1989 Dec;109(6 Pt 1):3073–3083. doi: 10.1083/jcb.109.6.3073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dodd J., Jessell T. M. Axon guidance and the patterning of neuronal projections in vertebrates. Science. 1988 Nov 4;242(4879):692–699. doi: 10.1126/science.3055291. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Euteneuer U., Schliwa M. Mechanism of centrosome positioning during the wound response in BSC-1 cells. J Cell Biol. 1992 Mar;116(5):1157–1166. doi: 10.1083/jcb.116.5.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Forscher P., Kaczmarek L. K., Buchanan J. A., Smith S. J. Cyclic AMP induces changes in distribution and transport of organelles within growth cones of Aplysia bag cell neurons. J Neurosci. 1987 Nov;7(11):3600–3611. doi: 10.1523/JNEUROSCI.07-11-03600.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Forscher P., Lin C. H., Thompson C. Novel form of growth cone motility involving site-directed actin filament assembly. Nature. 1992 Jun 11;357(6378):515–518. doi: 10.1038/357515a0. [DOI] [PubMed] [Google Scholar]
  19. Forscher P., Smith S. J. Actions of cytochalasins on the organization of actin filaments and microtubules in a neuronal growth cone. J Cell Biol. 1988 Oct;107(4):1505–1516. doi: 10.1083/jcb.107.4.1505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Goodman C. S., Bastiani M. J., Doe C. Q., du Lac S., Helfand S. L., Kuwada J. Y., Thomas J. B. Cell recognition during neuronal development. Science. 1984 Sep 21;225(4668):1271–1279. doi: 10.1126/science.6474176. [DOI] [PubMed] [Google Scholar]
  21. Goslin K., Birgbauer E., Banker G., Solomon F. The role of cytoskeleton in organizing growth cones: a microfilament-associated growth cone component depends upon microtubules for its localization. J Cell Biol. 1989 Oct;109(4 Pt 1):1621–1631. doi: 10.1083/jcb.109.4.1621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Griffith L. M., Pollard T. D. Evidence for actin filament-microtubule interaction mediated by microtubule-associated proteins. J Cell Biol. 1978 Sep;78(3):958–965. doi: 10.1083/jcb.78.3.958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Gundersen R. W., Barrett J. N. Characterization of the turning response of dorsal root neurites toward nerve growth factor. J Cell Biol. 1980 Dec;87(3 Pt 1):546–554. doi: 10.1083/jcb.87.3.546. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Heidemann S. R., Lamoureux P., Buxbaum R. E. Growth cone behavior and production of traction force. J Cell Biol. 1990 Nov;111(5 Pt 1):1949–1957. doi: 10.1083/jcb.111.5.1949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Hortsch M., Goodman C. S. Cell and substrate adhesion molecules in Drosophila. Annu Rev Cell Biol. 1991;7:505–557. doi: 10.1146/annurev.cb.07.110191.002445. [DOI] [PubMed] [Google Scholar]
  26. Jessell T. M. Adhesion molecules and the hierarchy of neural development. Neuron. 1988 Mar;1(1):3–13. doi: 10.1016/0896-6273(88)90204-8. [DOI] [PubMed] [Google Scholar]
  27. Kaczmarek L. K., Finbow M., Revel J. P., Strumwasser F. The morphology and coupling of Aplysia bag cells within the abdominal ganglion and in cell culture. J Neurobiol. 1979 Nov;10(6):535–550. doi: 10.1002/neu.480100604. [DOI] [PubMed] [Google Scholar]
  28. Kucik D. F., Elson E. L., Sheetz M. P. Forward transport of glycoproteins on leading lamellipodia in locomoting cells. Nature. 1989 Jul 27;340(6231):315–317. doi: 10.1038/340315a0. [DOI] [PubMed] [Google Scholar]
  29. Kuznetsov S. A., Langford G. M., Weiss D. G. Actin-dependent organelle movement in squid axoplasm. Nature. 1992 Apr 23;356(6371):722–725. doi: 10.1038/356722a0. [DOI] [PubMed] [Google Scholar]
  30. Lamoureux P., Buxbaum R. E., Heidemann S. R. Direct evidence that growth cones pull. Nature. 1989 Jul 13;340(6229):159–162. doi: 10.1038/340159a0. [DOI] [PubMed] [Google Scholar]
  31. Lamoureux P., Zheng J., Buxbaum R. E., Heidemann S. R. A cytomechanical investigation of neurite growth on different culture surfaces. J Cell Biol. 1992 Aug;118(3):655–661. doi: 10.1083/jcb.118.3.655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lasek R. J. Polymer sliding in axons. J Cell Sci Suppl. 1986;5:161–179. doi: 10.1242/jcs.1986.supplement_5.10. [DOI] [PubMed] [Google Scholar]
  33. Letourneau P. C., Shattuck T. A. Distribution and possible interactions of actin-associated proteins and cell adhesion molecules of nerve growth cones. Development. 1989 Mar;105(3):505–519. doi: 10.1242/dev.105.3.505. [DOI] [PubMed] [Google Scholar]
  34. Letourneau P. C., Shattuck T. A., Ressler A. H. "Pull" and "push" in neurite elongation: observations on the effects of different concentrations of cytochalasin B and taxol. Cell Motil Cytoskeleton. 1987;8(3):193–209. doi: 10.1002/cm.970080302. [DOI] [PubMed] [Google Scholar]
  35. Lewis A. K., Bridgman P. C. Nerve growth cone lamellipodia contain two populations of actin filaments that differ in organization and polarity. J Cell Biol. 1992 Dec;119(5):1219–1243. doi: 10.1083/jcb.119.5.1219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Marsh L., Letourneau P. C. Growth of neurites without filopodial or lamellipodial activity in the presence of cytochalasin B. J Cell Biol. 1984 Dec;99(6):2041–2047. doi: 10.1083/jcb.99.6.2041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Miller M., Bower E., Levitt P., Li D., Chantler P. D. Myosin II distribution in neurons is consistent with a role in growth cone motility but not synaptic vesicle mobilization. Neuron. 1992 Jan;8(1):25–44. doi: 10.1016/0896-6273(92)90106-n. [DOI] [PubMed] [Google Scholar]
  38. Mitchison T., Kirschner M. Cytoskeletal dynamics and nerve growth. Neuron. 1988 Nov;1(9):761–772. doi: 10.1016/0896-6273(88)90124-9. [DOI] [PubMed] [Google Scholar]
  39. Morales M., Fifkova E. Distribution of MAP2 in dendritic spines and its colocalization with actin. An immunogold electron-microscope study. Cell Tissue Res. 1989 Jun;256(3):447–456. doi: 10.1007/BF00225592. [DOI] [PubMed] [Google Scholar]
  40. O'Connor T. P., Duerr J. S., Bentley D. Pioneer growth cone steering decisions mediated by single filopodial contacts in situ. J Neurosci. 1990 Dec;10(12):3935–3946. doi: 10.1523/JNEUROSCI.10-12-03935.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Okabe S., Hirokawa N. Actin dynamics in growth cones. J Neurosci. 1991 Jul;11(7):1918–1929. doi: 10.1523/JNEUROSCI.11-07-01918.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Patel N., Poo M. M. Orientation of neurite growth by extracellular electric fields. J Neurosci. 1982 Apr;2(4):483–496. doi: 10.1523/JNEUROSCI.02-04-00483.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Peng H. B., Cheng P. C. Formation of postsynaptic specializations induced by latex beads in cultured muscle cells. J Neurosci. 1982 Dec;2(12):1760–1774. doi: 10.1523/JNEUROSCI.02-12-01760.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Peng H. B., Markey D. R., Muhlach W. L., Pollack E. D. Development of presynaptic specializations induced by basic polypeptide-coated latex beads in spinal cord cultures. Synapse. 1987;1(1):10–19. doi: 10.1002/syn.890010104. [DOI] [PubMed] [Google Scholar]
  45. Reichardt L. F., Tomaselli K. J. Extracellular matrix molecules and their receptors: functions in neural development. Annu Rev Neurosci. 1991;14:531–570. doi: 10.1146/annurev.ne.14.030191.002531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sabry J. H., O'Connor T. P., Evans L., Toroian-Raymond A., Kirschner M., Bentley D. Microtubule behavior during guidance of pioneer neuron growth cones in situ. J Cell Biol. 1991 Oct;115(2):381–395. doi: 10.1083/jcb.115.2.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Schliwa M., van Blerkom J. Structural interaction of cytoskeletal components. J Cell Biol. 1981 Jul;90(1):222–235. doi: 10.1083/jcb.90.1.222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Selden S. C., Pollard T. D. Phosphorylation of microtubule-associated proteins regulates their interaction with actin filaments. J Biol Chem. 1983 Jun 10;258(11):7064–7071. [PubMed] [Google Scholar]
  49. Sheetz M. P., Turney S., Qian H., Elson E. L. Nanometre-level analysis demonstrates that lipid flow does not drive membrane glycoprotein movements. Nature. 1989 Jul 27;340(6231):284–288. doi: 10.1038/340284a0. [DOI] [PubMed] [Google Scholar]
  50. Shpetner H. S., Vallee R. B. Identification of dynamin, a novel mechanochemical enzyme that mediates interactions between microtubules. Cell. 1989 Nov 3;59(3):421–432. doi: 10.1016/0092-8674(89)90027-5. [DOI] [PubMed] [Google Scholar]
  51. Smith S. J. Neuronal cytomechanics: the actin-based motility of growth cones. Science. 1988 Nov 4;242(4879):708–715. doi: 10.1126/science.3055292. [DOI] [PubMed] [Google Scholar]
  52. Takeichi M. Cadherin cell adhesion receptors as a morphogenetic regulator. Science. 1991 Mar 22;251(5000):1451–1455. doi: 10.1126/science.2006419. [DOI] [PubMed] [Google Scholar]
  53. Tanaka E. M., Kirschner M. W. Microtubule behavior in the growth cones of living neurons during axon elongation. J Cell Biol. 1991 Oct;115(2):345–363. doi: 10.1083/jcb.115.2.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Tessier-Lavigne M., Placzek M., Lumsden A. G., Dodd J., Jessell T. M. Chemotropic guidance of developing axons in the mammalian central nervous system. Nature. 1988 Dec 22;336(6201):775–778. doi: 10.1038/336775a0. [DOI] [PubMed] [Google Scholar]
  55. Theriot J. A., Mitchison T. J. Actin microfilament dynamics in locomoting cells. Nature. 1991 Jul 11;352(6331):126–131. doi: 10.1038/352126a0. [DOI] [PubMed] [Google Scholar]
  56. Theriot J. A., Mitchison T. J. Comparison of actin and cell surface dynamics in motile fibroblasts. J Cell Biol. 1992 Oct;119(2):367–377. doi: 10.1083/jcb.119.2.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Travis J. L., Bowser S. S. Microtubule-dependent reticulopodial motility: is there a role for actin? Cell Motil Cytoskeleton. 1986;6(2):146–152. doi: 10.1002/cm.970060212. [DOI] [PubMed] [Google Scholar]
  58. de Brabander M., Nuydens R., Ishihara A., Holifield B., Jacobson K., Geerts H. Lateral diffusion and retrograde movements of individual cell surface components on single motile cells observed with Nanovid microscopy. J Cell Biol. 1991 Jan;112(1):111–124. doi: 10.1083/jcb.112.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES