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. 1981 Jan 1;88(1):172–178. doi: 10.1083/jcb.88.1.172

Clathrin is axonally transported as part of slow component b: the microfilament complex

PMCID: PMC2111710  PMID: 6162851

Abstract

During axonal transport, membranes travel down axons at a rapid rate, whereas the cytoskeletal elements travel in either of two slow components, SCa (with tubulin and neurofilament protein) and SCb (with actin). Clathrin, the highly ordered, structural coat protein of coated vesicles, has recently been shown to be able to interact in vitro with cytoskeletal proteins in addition to membranes. The present study examines whether clathrin travels preferentially with the membrane elements or the cytoskeletal elements when it is axonally transported. Guinea pig visual system was labeled with tritiated amino acids. Radioactive SDS-polyacrylamide gel electrophoresis profiles from the major components of transport were coelectrophoresed with clathrin. Only SCb had a band comigrating with clathrin. In addition, radioactive clathrin was purified from guinea pig brain containing only radioactive SCb polypeptides. Kinetic analysis of the putative clathrin band in SCb revealed that it travels entirely within the SCb wave. Thus we conclude that clathrin travels preferentially with the cytoskeletal proteins making up SCb, rather than with the membranes and membrane-associated proteins in the fast component.

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Selected References

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

  1. Black M. M., Lasek R. J. Axonal transport of actin: slow component b is the principal source of actin for the axon. Brain Res. 1979 Aug 10;171(3):401–413. doi: 10.1016/0006-8993(79)91045-x. [DOI] [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. Blitz A. L., Fine R. E., Toselli P. A. Evidence that coated vesicles isolated from brain are calcium-sequestering organelles resembling sarcoplasmic reticulum. J Cell Biol. 1977 Oct;75(1):135–147. doi: 10.1083/jcb.75.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  5. Brimijoin S. Stop-flow: a new technique for measuring axonal transport, and its application to the transport of dopamine-beta-hydroxylase. J Neurobiol. 1975 Jul;6(4):379–394. doi: 10.1002/neu.480060404. [DOI] [PubMed] [Google Scholar]
  6. Cancalon P., Beidler L. M. Distribution along the axon and into various subcellular fractions of molecules labeled with (3H)leucine and rapidly transported in the garfish olfactory nerve. Brain Res. 1975 May 23;89(2):225–244. doi: 10.1016/0006-8993(75)90715-5. [DOI] [PubMed] [Google Scholar]
  7. Crowther R. A., Finch J. T., Pearse B. M. On the structure of coated vesicles. J Mol Biol. 1976 Jun 5;103(4):785–798. doi: 10.1016/0022-2836(76)90209-6. [DOI] [PubMed] [Google Scholar]
  8. Droz B., Rambourg A., Koenig H. L. The smooth endoplasmic reticulum: structure and role in the renewal of axonal membrane and synaptic vesicles by fast axonal transport. Brain Res. 1975 Jul 25;93(1):1–13. doi: 10.1016/0006-8993(75)90282-6. [DOI] [PubMed] [Google Scholar]
  9. Forman D. S., Grafstein B., McEwen B. S. Rapid axonal transport of ( 3 H)fucosyl glycoproteins in the goldfish optic system. Brain Res. 1972 Dec 24;48:327–342. doi: 10.1016/0006-8993(72)90187-4. [DOI] [PubMed] [Google Scholar]
  10. Forman D. S., Ledeen R. W. Axonal transport of gangliosides in the goldfish optic nerve. Science. 1972 Aug 18;177(4049):630–633. doi: 10.1126/science.177.4049.630. [DOI] [PubMed] [Google Scholar]
  11. Friend D. S., Farquhar M. G. Functions of coated vesicles during protein absorption in the rat vas deferens. J Cell Biol. 1967 Nov;35(2):357–376. doi: 10.1083/jcb.35.2.357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Goldstein J. L., Anderson R. G., Brown M. S. Coated pits, coated vesicles, and receptor-mediated endocytosis. Nature. 1979 Jun 21;279(5715):679–685. doi: 10.1038/279679a0. [DOI] [PubMed] [Google Scholar]
  13. Gorden P., Carpentier J. L., Cohen S., Orci L. Epidermal growth factor: morphological demonstration of binding, internalization, and lysosomal association in human fibroblasts. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5025–5029. doi: 10.1073/pnas.75.10.5025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heuser J. E., Reese T. S. Evidence for recycling of synaptic vesicle membrane during transmitter release at the frog neuromuscular junction. J Cell Biol. 1973 May;57(2):315–344. doi: 10.1083/jcb.57.2.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Heuser J. Three-dimensional visualization of coated vesicle formation in fibroblasts. J Cell Biol. 1980 Mar;84(3):560–583. doi: 10.1083/jcb.84.3.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Holtzman E. The origin and fate of secretory packages, especially synaptic vesicles. Neuroscience. 1977;2(3):327–355. doi: 10.1016/0306-4522(77)90001-x. [DOI] [PubMed] [Google Scholar]
  18. Keen J. H., Willingham M. C., Pastan I. H. Clathrin-coated vesicles: isolation, dissociation and factor-dependent reassociation of clathrin baskets. Cell. 1979 Feb;16(2):303–312. doi: 10.1016/0092-8674(79)90007-2. [DOI] [PubMed] [Google Scholar]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Lasek R. J., Gainer H., Przybylski R. J. Transfer of newly synthesized proteins from Schwann cells to the squid giant axon. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1188–1192. doi: 10.1073/pnas.71.4.1188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  22. Neville D. M., Jr Molecular weight determination of protein-dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer system. J Biol Chem. 1971 Oct 25;246(20):6328–6334. [PubMed] [Google Scholar]
  23. 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]
  24. Ockleford C. D., Whyte A. Differeniated regions of human placental cell surface associated with exchange of materials between maternal and foetal blood: coated vesicles. J Cell Sci. 1977 Jun;25:293–312. doi: 10.1242/jcs.25.1.293. [DOI] [PubMed] [Google Scholar]
  25. Pearse B. M. Clathrin: a unique protein associated with intracellular transfer of membrane by coated vesicles. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1255–1259. doi: 10.1073/pnas.73.4.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pearse B. M. Coated vesicles from pig brain: purification and biochemical characterization. J Mol Biol. 1975 Sep 5;97(1):93–98. doi: 10.1016/s0022-2836(75)80024-6. [DOI] [PubMed] [Google Scholar]
  27. Rothman J. E., Fine R. E. Coated vesicles transport newly synthesized membrane glycoproteins from endoplasmic reticulum to plasma membrane in two successive stages. Proc Natl Acad Sci U S A. 1980 Feb;77(2):780–784. doi: 10.1073/pnas.77.2.780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schook W., Puszkin S., Bloom W., Ores C., Kochwa S. Mechanochemical properties of brain clathrin: interactions with actin and alpha-actinin and polymerization into basketlike structures or filaments. Proc Natl Acad Sci U S A. 1979 Jan;76(1):116–120. doi: 10.1073/pnas.76.1.116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schwartz J. H., Goldman J. E., Ambron R. T., Goldberg D. J. Axonal transport of vesicles carrying (3H)serotonin in the metacerebral neuron of Aplysia californica. Cold Spring Harb Symp Quant Biol. 1976;40:83–92. doi: 10.1101/sqb.1976.040.01.010. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Willard M. The identification of two intra-axonally transported polypeptides resembling myosin in some respects in the rabbit visual system. J Cell Biol. 1977 Oct;75(1):1–11. doi: 10.1083/jcb.75.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Willard M., Wiseman M., Levine J., Skene P. Axonal transport of actin in rabbit retinal ganglion cells. J Cell Biol. 1979 Jun;81(3):581–591. doi: 10.1083/jcb.81.3.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Woods J. W., Woodward M. P., Roth T. F. Common features of coated vesicles from dissimilar tissues: composition and structure. J Cell Sci. 1978 Apr;30:87–97. doi: 10.1242/jcs.30.1.87. [DOI] [PubMed] [Google Scholar]
  34. Woodward M. P., Roth T. F. Coated vesicles: characterization, selective dissociation, and reassembly. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4394–4398. doi: 10.1073/pnas.75.9.4394. [DOI] [PMC free article] [PubMed] [Google Scholar]

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