Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1987 Dec 1;105(6):2827–2835. doi: 10.1083/jcb.105.6.2827

Rapidly transported organelles containing membrane and cytoskeletal components: their relation to axonal growth

PMCID: PMC2114729  PMID: 3693400

Abstract

We have examined the movements, composition, and cellular origin of phase-dense varicosities in cultures of chick sympathetic and sensory neurons. These organelles are variable in diameter (typically between 0.2 and 2 microns) and undergo saltatory movements both towards and away from the neuronal cell body. Their mean velocities vary inversely with the size of the organelle and are greater in the retrograde than the anterograde direction. Organelles stain with the lipophilic dye 1, 1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine and with antibodies to cytoskeletal components. In cultures double-stained with antibodies to alpha-tubulin and 70-kD neurofilament protein (NF-L), approximately 40% of the organelles stain for tubulin, 30% stain for NF- L, 10% stain for both tubulin and NF-L, and 40% show no staining with either antibody. The association of cytoskeletal proteins with the organelles shows that these proteins are able to move by a form of rapid axonal transport. Under most culture conditions the predominant direction of movement is towards the cell body, suggesting that the organelles are produced at or near the growth cone. Retrograde movements continue in culture medium lacking protein or high molecular mass components and increase under conditions in which the advance of the growth cone is arrested. There is a fourfold increase in the number of organelles moving retrogradely in neurites that encounter a substratum-associated barrier to elongation; retrograde movements increase similarly in cultures exposed to cytochalasin at levels known to block growth cone advance. No previously described organelle shows behavior coordinated with axonal growth in this way. We propose that the organelles contain membrane and cytoskeletal components that have been delivered to the growth cone, by slow or fast anterograde transport, in excess of the amounts required to synthesize more axon. In view of their rapid mobility and variable contents, we suggest that they be called "neuronal parcels."

Full Text

The Full Text of this article is available as a PDF (2.7 MB).

Selected References

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

  1. Axelrod D. Carbocyanine dye orientation in red cell membrane studied by microscopic fluorescence polarization. Biophys J. 1979 Jun;26(3):557–573. doi: 10.1016/S0006-3495(79)85271-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Brown S., Levinson W., Spudich J. A. Cytoskeletal elements of chick embryo fibroblasts revealed by detergent extraction. J Supramol Struct. 1976;5(2):119–130. doi: 10.1002/jss.400050203. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Heriot K., Gambetti P., Lasek R. J. Proteins transported in slow components a and b of axonal transport are distributed differently in the transverse plane of the axon. J Cell Biol. 1985 Apr;100(4):1167–1172. doi: 10.1083/jcb.100.4.1167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hoffman P. N., Lasek R. J. The slow component of axonal transport. Identification of major structural polypeptides of the axon and their generality among mammalian neurons. J Cell Biol. 1975 Aug;66(2):351–366. doi: 10.1083/jcb.66.2.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hollenbeck P. J., Bray D., Adams R. J. Effects of the uncoupling agents FCCP and CCCP on the saltatory movements of cytoplasmic organelles. Cell Biol Int Rep. 1985 Feb;9(2):193–199. doi: 10.1016/0309-1651(85)90094-3. [DOI] [PubMed] [Google Scholar]
  8. Honig M. G., Hume R. I. Fluorescent carbocyanine dyes allow living neurons of identified origin to be studied in long-term cultures. J Cell Biol. 1986 Jul;103(1):171–187. doi: 10.1083/jcb.103.1.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jacobs M., Choo Q. L., Thomas C. Vimentin and 70K neurofilament protein co-exist in embryonic neurones from spinal ganglia. J Neurochem. 1982 Apr;38(4):969–977. doi: 10.1111/j.1471-4159.1982.tb05337.x. [DOI] [PubMed] [Google Scholar]
  10. Jacobson K., Hou Y., Derzko Z., Wojcieszyn J., Organisciak D. Lipid lateral diffusion in the surface membrane of cells and in multibilayers formed from plasma membrane lipids. Biochemistry. 1981 Sep 1;20(18):5268–5275. doi: 10.1021/bi00521a027. [DOI] [PubMed] [Google Scholar]
  11. Johnson G. D., Nogueira Araujo G. M. A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods. 1981;43(3):349–350. doi: 10.1016/0022-1759(81)90183-6. [DOI] [PubMed] [Google Scholar]
  12. Kilmartin J. V., Wright B., Milstein C. Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line. J Cell Biol. 1982 Jun;93(3):576–582. doi: 10.1083/jcb.93.3.576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Koenig E., Kinsman S., Repasky E., Sultz L. Rapid mobility of motile varicosities and inclusions containing alpha-spectrin, actin, and calmodulin in regenerating axons in vitro. J Neurosci. 1985 Mar;5(3):715–729. doi: 10.1523/JNEUROSCI.05-03-00715.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lander A. D., Fujii D. K., Gospodarowicz D., Reichardt L. F. Characterization of a factor that promotes neurite outgrowth: evidence linking activity to a heparan sulfate proteoglycan. J Cell Biol. 1982 Sep;94(3):574–585. doi: 10.1083/jcb.94.3.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Ludueña M. A. The growth of spinal ganglion neurons in serum-free medium. Dev Biol. 1973 Aug;33(2):470–476. doi: 10.1016/0012-1606(73)90152-8. [DOI] [PubMed] [Google Scholar]
  17. McGee-Russell S. M. Dynamic activities and labile microtubules in cytoplasmic transport in the marine foraminiferan Allogromia. Symp Soc Exp Biol. 1974;(28):157–189. [PubMed] [Google Scholar]
  18. Nixon R. A., Logvinenko K. B. Multiple fates of newly synthesized neurofilament proteins: evidence for a stationary neurofilament network distributed nonuniformly along axons of retinal ganglion cell neurons. J Cell Biol. 1986 Feb;102(2):647–659. doi: 10.1083/jcb.102.2.647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Roots B. I. Neurofilament accumulation induced in synapses by leupeptin. Science. 1983 Sep 2;221(4614):971–972. doi: 10.1126/science.6192501. [DOI] [PubMed] [Google Scholar]
  20. Schlessinger J., Axelrod D., Koppel D. E., Webb W. W., Elson E. L. Lateral transport of a lipid probe and labeled proteins on a cell membrane. Science. 1977 Jan 21;195(4275):307–309. doi: 10.1126/science.556653. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Weber K., Rathke P. C., Osborn M. Cytoplasmic microtubular images in glutaraldehyde-fixed tissue culture cells by electron microscopy and by immunofluorescence microscopy. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1820–1824. doi: 10.1073/pnas.75.4.1820. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES