Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1985 May 1;100(5):1736–1752. doi: 10.1083/jcb.100.5.1736

Gliding movement of and bidirectional transport along single native microtubules from squid axoplasm: evidence for an active role of microtubules in cytoplasmic transport

PMCID: PMC2113850  PMID: 2580845

Abstract

Native microtubules prepared from extruded and dissociated axoplasm have been observed to transport organelles and vesicles unidirectionally in fresh preparations and more slowly and bidirectionally in older preparations. Both endogenous and exogenous (fluorescent polystyrene) particles in rapid Brownian motion alight on and adhere to microtubules and are transported along them. Particles can switch from one intersecting microtubule to another and move in either direction. Microtubular segments 1 to 30 microns long, produced by gentle homogenization, glide over glass surfaces for hundreds of micrometers in straight lines unless acted upon by obstacles. While gliding they transport particles either in the same (forward) direction and/or in the backward direction. Particle movement and gliding of microtubule segments require ATP and are insensitive to taxol (30 microM). It appears, therefore, that the mechanisms producing the motive force are very closely associated with the native microtubule itself or with its associated proteins. Although these movements appear irreconcilable with several current theories of fast axoplasmic transport, in this article we propose two models that might explain the observed phenomena and, by extension, the process of fast axoplasmic transport itself. The findings presented and the possible mechanisms proposed for fast axoplasmic transport have potential applications across the spectrum of microtubule-based motility processes.

Full Text

The Full Text of this article is available as a PDF (2.0 MB).

Selected References

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

  1. Adams R. J., Bray D. Rapid transport of foreign particles microinjected into crab axons. Nature. 1983 Jun 23;303(5919):718–720. doi: 10.1038/303718a0. [DOI] [PubMed] [Google Scholar]
  2. Adams R. J. Organelle movement in axons depends on ATP. Nature. 1982 May 27;297(5864):327–329. doi: 10.1038/297327a0. [DOI] [PubMed] [Google Scholar]
  3. Allen R. D., Allen N. S., Travis J. L. Video-enhanced contrast, differential interference contrast (AVEC-DIC) microscopy: a new method capable of analyzing microtubule-related motility in the reticulopodial network of Allogromia laticollaris. Cell Motil. 1981;1(3):291–302. doi: 10.1002/cm.970010303. [DOI] [PubMed] [Google Scholar]
  4. Allen R. D., Allen N. S. Video-enhanced microscopy with a computer frame memory. J Microsc. 1983 Jan;129(Pt 1):3–17. doi: 10.1111/j.1365-2818.1983.tb04157.x. [DOI] [PubMed] [Google Scholar]
  5. Allen R. D., Metuzals J., Tasaki I., Brady S. T., Gilbert S. P. Fast axonal transport in squid giant axon. Science. 1982 Dec 10;218(4577):1127–1129. doi: 10.1126/science.6183744. [DOI] [PubMed] [Google Scholar]
  6. Allen R. D. New observations on cell architecture and dynamics by video-enhanced contrast optical microscopy. Annu Rev Biophys Biophys Chem. 1985;14:265–290. doi: 10.1146/annurev.bb.14.060185.001405. [DOI] [PubMed] [Google Scholar]
  7. Allen R. D., Travis J. L., Allen N. S., Yilmaz H. Video-enhanced contrast polarization (AVEC-POL) microscopy: a new method applied to the detection of birefringence in the motile reticulopodial network of Allogromia laticollaris. Cell Motil. 1981;1(3):275–289. doi: 10.1002/cm.970010302. [DOI] [PubMed] [Google Scholar]
  8. Bajer A. S., Cypher C., Molè-Bajer J., Howard H. M. Taxol-induced anaphase reversal: evidence that elongating microtubules can exert a pushing force in living cells. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6569–6573. doi: 10.1073/pnas.79.21.6569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Beckerle M. C. Microinjected fluorescent polystyrene beads exhibit saltatory motion in tissue culture cells. J Cell Biol. 1984 Jun;98(6):2126–2132. doi: 10.1083/jcb.98.6.2126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Beckerle M. C., Porter K. R. Inhibitors of dynein activity block intracellular transport in erythrophores. Nature. 1982 Feb 25;295(5851):701–703. doi: 10.1038/295701a0. [DOI] [PubMed] [Google Scholar]
  11. Black M. M., Lasek R. J. Slow components of axonal transport: two cytoskeletal networks. J Cell Biol. 1980 Aug;86(2):616–623. doi: 10.1083/jcb.86.2.616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Brady S. T., Lasek R. J., Allen R. D. Fast axonal transport in extruded axoplasm from squid giant axon. Science. 1982 Dec 10;218(4577):1129–1131. doi: 10.1126/science.6183745. [DOI] [PubMed] [Google Scholar]
  13. Brady S. T., Lasek R. J., Allen R. D. Video microscopy of fast axonal transport in extruded axoplasm: a new model for study of molecular mechanisms. Cell Motil. 1985;5(2):81–101. doi: 10.1002/cm.970050203. [DOI] [PubMed] [Google Scholar]
  14. Brady S. T., Lasek R. J., Allen R. D., Yin H. L., Stossel T. P. Gelsolin inhibition of fast axonal transport indicates a requirement for actin microfilaments. Nature. 1984 Jul 5;310(5972):56–58. doi: 10.1038/310056a0. [DOI] [PubMed] [Google Scholar]
  15. Buckley I., Stewart M. Ciliary but not saltatory movements are inhibited by vanadate microinjected into living cultured cells. Cell Motil. 1983;3(2):167–184. doi: 10.1002/cm.970030206. [DOI] [PubMed] [Google Scholar]
  16. Burton P. R., Paige J. L. Polarity of axoplasmic microtubules in the olfactory nerve of the frog. Proc Natl Acad Sci U S A. 1981 May;78(5):3269–3273. doi: 10.1073/pnas.78.5.3269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Collins F. Axon initiation by ciliary neurons in culture. Dev Biol. 1978 Jul;65(1):50–57. doi: 10.1016/0012-1606(78)90178-1. [DOI] [PubMed] [Google Scholar]
  18. Forman D. S., Brown K. J., Livengood D. R. Fast axonal transport in permeabilized lobster giant axons is inhibited by vanadate. J Neurosci. 1983 Jun;3(6):1279–1288. doi: 10.1523/JNEUROSCI.03-06-01279.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Forman D. S. Vanadate inhibits saltatory organelle movement in a permeabilized cell model. Exp Cell Res. 1982 Sep;141(1):139–147. doi: 10.1016/0014-4827(82)90076-3. [DOI] [PubMed] [Google Scholar]
  20. Goldberg D. J., Harris D. A., Lubit B. W., Schwartz J. H. Analysis of the mechanism of fast axonal transport by intracellular injection of potentially inhibitory macromolecules: evidence for a possible role of actin filaments. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7448–7452. doi: 10.1073/pnas.77.12.7448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Goldberg D. J. Microinjection into an identified axon to study the mechanism of fast axonal transport. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4818–4822. doi: 10.1073/pnas.79.15.4818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Grafstein B., Forman D. S. Intracellular transport in neurons. Physiol Rev. 1980 Oct;60(4):1167–1283. doi: 10.1152/physrev.1980.60.4.1167. [DOI] [PubMed] [Google Scholar]
  23. Hayden J. H., Allen R. D. Detection of single microtubules in living cells: particle transport can occur in both directions along the same microtubule. J Cell Biol. 1984 Nov;99(5):1785–1793. doi: 10.1083/jcb.99.5.1785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hayden J. H., Allen R. D., Goldman R. D. Cytoplasmic transport in keratocytes: direct visualization of particle translocation along microtubules. Cell Motil. 1983;3(1):1–19. doi: 10.1002/cm.970030102. [DOI] [PubMed] [Google Scholar]
  25. Heidemann S. R., Landers J. M., Hamborg M. A. Polarity orientation of axonal microtubules. J Cell Biol. 1981 Dec;91(3 Pt 1):661–665. doi: 10.1083/jcb.91.3.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Heslop J. P. Fast transport along nerves. Symp Soc Exp Biol. 1974;(28):209–227. [PubMed] [Google Scholar]
  27. Hodge A. J., Adelman W. J., Jr The neuroplasmic network in Loligo and Hermissenda neurons. J Ultrastruct Res. 1980 Feb;70(2):220–241. doi: 10.1016/s0022-5320(80)80007-4. [DOI] [PubMed] [Google Scholar]
  28. Hollenbeck P. J., Suprynowicz F., Cande W. Z. Cytoplasmic dynein-like ATPase cross-links microtubules in an ATP-sensitive manner. J Cell Biol. 1984 Oct;99(4 Pt 1):1251–1258. doi: 10.1083/jcb.99.4.1251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Isenberg G., Schubert P., Kreutzberg G. W. Experimental approach to test the role of actin in axonal transport. Brain Res. 1980 Aug 4;194(2):588–593. doi: 10.1016/0006-8993(80)91247-0. [DOI] [PubMed] [Google Scholar]
  30. Jarosch R., Foissner I. A rotation model for microtubule and filament sliding. Eur J Cell Biol. 1982 Feb;26(2):295–302. [PubMed] [Google Scholar]
  31. Kerkut G. A. Axoplasmic transport. Comp Biochem Physiol A Comp Physiol. 1975 Aug 1;51(4):701–704. doi: 10.1016/0300-9629(75)90041-9. [DOI] [PubMed] [Google Scholar]
  32. Langford G. M. Length and appearance of projections on neuronal microtubules in vitro after negative staining: evidence against a crosslinking function for MAPs. J Ultrastruct Res. 1983 Oct;85(1):1–10. doi: 10.1016/s0022-5320(83)90111-9. [DOI] [PubMed] [Google Scholar]
  33. Lasek R. J., Garner J. A., Brady S. T. Axonal transport of the cytoplasmic matrix. J Cell Biol. 1984 Jul;99(1 Pt 2):212s–221s. doi: 10.1083/jcb.99.1.212s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Margolis R. L., Wilson L. Microtubule treadmills--possible molecular machinery. Nature. 1981 Oct 29;293(5835):705–711. doi: 10.1038/293705a0. [DOI] [PubMed] [Google Scholar]
  35. Martz D., Lasek R. J., Brady S. T., Allen R. D. Mitochondrial motility in axons: membranous organelles may interact with the force generating system through multiple surface binding sites. Cell Motil. 1984;4(2):89–101. doi: 10.1002/cm.970040203. [DOI] [PubMed] [Google Scholar]
  36. Murphy D. B., Hiebsch R. R., Wallis K. T. Identity and Origin of the ATPase activity associated with neuronal microtubules. I. The ATPase activity is associated with membrane vesicles. J Cell Biol. 1983 May;96(5):1298–1305. doi: 10.1083/jcb.96.5.1298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ochs S. Fast transport of materials in mammalian nerve fibers. Science. 1972 Apr 21;176(4032):252–260. doi: 10.1126/science.176.4032.252. [DOI] [PubMed] [Google Scholar]
  38. Rebhun L. I. Polarized intracellular particle transport: saltatory movements and cytoplasmic streaming. Int Rev Cytol. 1972;32:93–137. doi: 10.1016/s0074-7696(08)60339-3. [DOI] [PubMed] [Google Scholar]
  39. Sasaki S., Stevens J. K., Bodick N. Serial reconstruction of microtubular arrays within dendrites of the cat retinal ganglion cell: the cytoskeleton of a vertebrate dendrite. Brain Res. 1983 Jan 24;259(2):193–206. doi: 10.1016/0006-8993(83)91250-7. [DOI] [PubMed] [Google Scholar]
  40. Satir P., Wais-Steider J., Lebduska S., Nasr A., Avolio J. The mechanochemical cycle of the dynein arm. Cell Motil. 1981;1(3):303–327. doi: 10.1002/cm.970010304. [DOI] [PubMed] [Google Scholar]
  41. Schnapp B. J., Reese T. S. Cytoplasmic structure in rapid-frozen axons. J Cell Biol. 1982 Sep;94(3):667–669. doi: 10.1083/jcb.94.3.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schnapp B. J., Vale R. D., Sheetz M. P., Reese T. S. Single microtubules from squid axoplasm support bidirectional movement of organelles. Cell. 1985 Feb;40(2):455–462. doi: 10.1016/0092-8674(85)90160-6. [DOI] [PubMed] [Google Scholar]
  43. Schwartz J. H. Axonal transport: components, mechanisms, and specificity. Annu Rev Neurosci. 1979;2:467–504. doi: 10.1146/annurev.ne.02.030179.002343. [DOI] [PubMed] [Google Scholar]
  44. 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]
  45. Shimizu H. Dynamic cooperativity of molecular processes in active streaming, muscle contraction, and subcellular dynamics: the molecular mechanism of self-organization at the subcellular level. Adv Biophys. 1979;13:195–278. [PubMed] [Google Scholar]
  46. Smith D. S., Järlfors U., Beránek R. The organization of synaptic axcplasm in the lamprey (petromyzon marinus) central nervous system. J Cell Biol. 1970 Aug;46(2):199–219. doi: 10.1083/jcb.46.2.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tsukita S., Ishikawa H. The movement of membranous organelles in axons. Electron microscopic identification of anterogradely and retrogradely transported organelles. J Cell Biol. 1980 Mar;84(3):513–530. doi: 10.1083/jcb.84.3.513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Tytell M., Black M. M., Garner J. A., Lasek R. J. Axonal transport: each major rate component reflects the movement of distinct macromolecular complexes. Science. 1981 Oct 9;214(4517):179–181. doi: 10.1126/science.6169148. [DOI] [PubMed] [Google Scholar]
  49. Vale R. D., Schnapp B. J., Reese T. S., Sheetz M. P. Movement of organelles along filaments dissociated from the axoplasm of the squid giant axon. Cell. 1985 Feb;40(2):449–454. doi: 10.1016/0092-8674(85)90159-x. [DOI] [PubMed] [Google Scholar]
  50. Wolosewick J. J., Porter K. R. Microtrabecular lattice of the cytoplasmic ground substance. Artifact or reality. J Cell Biol. 1979 Jul;82(1):114–139. doi: 10.1083/jcb.82.1.114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wu T. Y. Hydrodynamics of swimming at low Reynolds numbers. Fortschr Zool. 1977;24(2-3):149–169. [PubMed] [Google Scholar]

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

RESOURCES