Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Nov 1;135(3):797–808. doi: 10.1083/jcb.135.3.797

Synaptic vesicles have two distinct recycling pathways

PMCID: PMC2121054  PMID: 8909551

Abstract

In this paper, evidence is presented that two distinct synaptic vesicle recycling pathways exist within a single terminal. One pathway emanates from the active zone, has a fast time course, involves no intermediate structures, and is blocked by exposure to high Mg2+/low Ca2+ saline, while the second pathway emanates at sites away from the active zone, has a slower time course, involves an endosomal intermediate, and is not sensitive to high Mg2+/low Ca2+. To visualize these two recycling pathways, the temperature-sensitive Drosophila mutant, shibire, in which vesicle recycling is normal at 19 degrees C but is blocked at 29 degrees C, was used. With exposure to 29 degrees C, complete vesicle depletion occurs as exocytosis proceeds while endocytosis is blocked. When the temperature is lowered to 26 degrees C, vesicle recycling membrane begins to accumulate as invaginations of the plasmalemma, but pinch-off is blocked. Under these experimental conditions, it was possible to distinguish the two separate pathways by electron microscopic analysis. These two pathways were further characterized by observing the normal recycling process at the permissive temperature, 19 degrees C. It is suggested that the function of these two recycling pathways might be to produce two distinct vesicle populations: the active zone and nonactive zone populations. The possibility that these two populations have different release characteristics and functions is discussed.

Full Text

The Full Text of this article is available as a PDF (6.1 MB).

Selected References

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

  1. Agoston D. V., Kosh J. W., Lisziewicz J., Whittaker V. P. Separation of recycling and reserve synaptic vesicles from cholinergic nerve terminals of the myenteric plexus of guinea pig ileum. J Neurochem. 1985 Jan;44(1):299–305. doi: 10.1111/j.1471-4159.1985.tb07144.x. [DOI] [PubMed] [Google Scholar]
  2. Bauerfeind R., Régnier-Vigouroux A., Flatmark T., Huttner W. B. Selective storage of acetylcholine, but not catecholamines, in neuroendocrine synaptic-like microvesicles of early endosomal origin. Neuron. 1993 Jul;11(1):105–121. doi: 10.1016/0896-6273(93)90275-v. [DOI] [PubMed] [Google Scholar]
  3. Betz W. J., Bewick G. S. Optical analysis of synaptic vesicle recycling at the frog neuromuscular junction. Science. 1992 Jan 10;255(5041):200–203. doi: 10.1126/science.1553547. [DOI] [PubMed] [Google Scholar]
  4. Betz W. J., Bewick G. S. Optical monitoring of transmitter release and synaptic vesicle recycling at the frog neuromuscular junction. J Physiol. 1993 Jan;460:287–309. doi: 10.1113/jphysiol.1993.sp019472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cameron P. L., Südhof T. C., Jahn R., De Camilli P. Colocalization of synaptophysin with transferrin receptors: implications for synaptic vesicle biogenesis. J Cell Biol. 1991 Oct;115(1):151–164. doi: 10.1083/jcb.115.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ceccarelli B., Grohovaz F., Hurlbut W. P. Freeze-fracture studies of frog neuromuscular junctions during intense release of neurotransmitter. I. Effects of black widow spider venom and Ca2+-free solutions on the structure of the active zone. J Cell Biol. 1979 Apr;81(1):163–177. doi: 10.1083/jcb.81.1.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ceccarelli B., Hurlbut W. P., Mauro A. Turnover of transmitter and synaptic vesicles at the frog neuromuscular junction. J Cell Biol. 1973 May;57(2):499–524. doi: 10.1083/jcb.57.2.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ceccarelli B., Hurlbut W. P. Vesicle hypothesis of the release of quanta of acetylcholine. Physiol Rev. 1980 Apr;60(2):396–441. doi: 10.1152/physrev.1980.60.2.396. [DOI] [PubMed] [Google Scholar]
  9. Chen M. S., Obar R. A., Schroeder C. C., Austin T. W., Poodry C. A., Wadsworth S. C., Vallee R. B. Multiple forms of dynamin are encoded by shibire, a Drosophila gene involved in endocytosis. Nature. 1991 Jun 13;351(6327):583–586. doi: 10.1038/351583a0. [DOI] [PubMed] [Google Scholar]
  10. Clift-O'Grady L., Linstedt A. D., Lowe A. W., Grote E., Kelly R. B. Biogenesis of synaptic vesicle-like structures in a pheochromocytoma cell line PC-12. J Cell Biol. 1990 May;110(5):1693–1703. doi: 10.1083/jcb.110.5.1693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Damke H., Baba T., Warnock D. E., Schmid S. L. Induction of mutant dynamin specifically blocks endocytic coated vesicle formation. J Cell Biol. 1994 Nov;127(4):915–934. doi: 10.1083/jcb.127.4.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fesce R., Grohovaz F., Valtorta F., Meldolesi J. Neurotransmitter release: fusion or 'kiss-and-run'? Trends Cell Biol. 1994 Jan;4(1):1–4. doi: 10.1016/0962-8924(94)90025-6. [DOI] [PubMed] [Google Scholar]
  13. Giompres P. E., Zimmermann H., Whittaker V. P. Purification of small dense vesicles from stimulated Torpedo electric tissue by glass bead column chromatography. Neuroscience. 1981;6(4):765–774. doi: 10.1016/0306-4522(81)90160-3. [DOI] [PubMed] [Google Scholar]
  14. Haimann C., Torri-Tarelli F., Fesce R., Ceccarelli B. Measurement of quantal secretion induced by ouabain and its correlation with depletion of synaptic vesicles. J Cell Biol. 1985 Nov;101(5 Pt 1):1953–1965. doi: 10.1083/jcb.101.5.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Heuser J. E. Review of electron microscopic evidence favouring vesicle exocytosis as the structural basis for quantal release during synaptic transmission. Q J Exp Physiol. 1989 Dec;74(7):1051–1069. doi: 10.1113/expphysiol.1989.sp003333. [DOI] [PubMed] [Google Scholar]
  17. Hinshaw J. E., Schmid S. L. Dynamin self-assembles into rings suggesting a mechanism for coated vesicle budding. Nature. 1995 Mar 9;374(6518):190–192. doi: 10.1038/374190a0. [DOI] [PubMed] [Google Scholar]
  18. Koenig J. H., Ikeda K. Disappearance and reformation of synaptic vesicle membrane upon transmitter release observed under reversible blockage of membrane retrieval. J Neurosci. 1989 Nov;9(11):3844–3860. doi: 10.1523/JNEUROSCI.09-11-03844.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Koenig J. H., Kosaka T., Ikeda K. The relationship between the number of synaptic vesicles and the amount of transmitter released. J Neurosci. 1989 Jun;9(6):1937–1942. doi: 10.1523/JNEUROSCI.09-06-01937.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Koenig J. H., Yamaoka K., Ikeda K. Calcium-induced translocation of synaptic vesicles to the active site. J Neurosci. 1993 Jun;13(6):2313–2322. doi: 10.1523/JNEUROSCI.13-06-02313.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kosaka T., Ikeda K. Possible temperature-dependent blockage of synaptic vesicle recycling induced by a single gene mutation in Drosophila. J Neurobiol. 1983 May;14(3):207–225. doi: 10.1002/neu.480140305. [DOI] [PubMed] [Google Scholar]
  22. Kosaka T., Ikeda K. Reversible blockage of membrane retrieval and endocytosis in the garland cell of the temperature-sensitive mutant of Drosophila melanogaster, shibirets1. J Cell Biol. 1983 Aug;97(2):499–507. doi: 10.1083/jcb.97.2.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kristensen H. K., Lau Y. Y., Ewing A. G. Capillary electrophoresis of single cells: observation of two compartments of neurotransmitter vesicles. J Neurosci Methods. 1994 Mar;51(2):183–188. doi: 10.1016/0165-0270(94)90009-4. [DOI] [PubMed] [Google Scholar]
  24. Pieribone V. A., Shupliakov O., Brodin L., Hilfiker-Rothenfluh S., Czernik A. J., Greengard P. Distinct pools of synaptic vesicles in neurotransmitter release. Nature. 1995 Jun 8;375(6531):493–497. doi: 10.1038/375493a0. [DOI] [PubMed] [Google Scholar]
  25. Prado M. A., Gomez M. V., Collier B. Mobilization of the readily releasable pool of acetylcholine from a sympathetic ganglion by tityustoxin in the presence of vesamicol. J Neurochem. 1992 Aug;59(2):544–552. doi: 10.1111/j.1471-4159.1992.tb09404.x. [DOI] [PubMed] [Google Scholar]
  26. Reetz A., Solimena M., Matteoli M., Folli F., Takei K., De Camilli P. GABA and pancreatic beta-cells: colocalization of glutamic acid decarboxylase (GAD) and GABA with synaptic-like microvesicles suggests their role in GABA storage and secretion. EMBO J. 1991 May;10(5):1275–1284. doi: 10.1002/j.1460-2075.1991.tb08069.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Régnier-Vigouroux A., Tooze S. A., Huttner W. B. Newly synthesized synaptophysin is transported to synaptic-like microvesicles via constitutive secretory vesicles and the plasma membrane. EMBO J. 1991 Dec;10(12):3589–3601. doi: 10.1002/j.1460-2075.1991.tb04925.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schwarz T. L. Genetic analysis of neurotransmitter release at the synapse. Curr Opin Neurobiol. 1994 Oct;4(5):633–639. doi: 10.1016/0959-4388(94)90003-5. [DOI] [PubMed] [Google Scholar]
  29. Takei K., McPherson P. S., Schmid S. L., De Camilli P. Tubular membrane invaginations coated by dynamin rings are induced by GTP-gamma S in nerve terminals. Nature. 1995 Mar 9;374(6518):186–190. doi: 10.1038/374186a0. [DOI] [PubMed] [Google Scholar]
  30. Thomas-Reetz A. C., De Camilli P. A role for synaptic vesicles in non-neuronal cells: clues from pancreatic beta cells and from chromaffin cells. FASEB J. 1994 Feb;8(2):209–216. doi: 10.1096/fasebj.8.2.7907072. [DOI] [PubMed] [Google Scholar]
  31. Torri-Tarelli F., Villa A., Valtorta F., De Camilli P., Greengard P., Ceccarelli B. Redistribution of synaptophysin and synapsin I during alpha-latrotoxin-induced release of neurotransmitter at the neuromuscular junction. J Cell Biol. 1990 Feb;110(2):449–459. doi: 10.1083/jcb.110.2.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Valtorta F., Fesce R., Grohovaz F., Haimann C., Hurlbut W. P., Iezzi N., Torri Tarelli F., Villa A., Ceccarelli B. Neurotransmitter release and synaptic vesicle recycling. Neuroscience. 1990;35(3):477–489. doi: 10.1016/0306-4522(90)90323-v. [DOI] [PubMed] [Google Scholar]
  33. Zimmermann H., Denston C. R. Separation of synaptic vesicles of different functional states from the cholinergic synapses of the Torpedo electric organ. Neuroscience. 1977;2(5):715–730. doi: 10.1016/0306-4522(77)90025-2. [DOI] [PubMed] [Google Scholar]
  34. Zimmermann H., Whittaker V. P. Morphological and biochemical heterogeneity of cholinergic synaptic vesicles. Nature. 1977 Jun 16;267(5612):633–635. doi: 10.1038/267633a0. [DOI] [PubMed] [Google Scholar]
  35. van der Bliek A. M., Meyerowitz E. M. Dynamin-like protein encoded by the Drosophila shibire gene associated with vesicular traffic. Nature. 1991 May 30;351(6325):411–414. doi: 10.1038/351411a0. [DOI] [PubMed] [Google Scholar]
  36. von Gersdorff H., Matthews G. Dynamics of synaptic vesicle fusion and membrane retrieval in synaptic terminals. Nature. 1994 Feb 24;367(6465):735–739. doi: 10.1038/367735a0. [DOI] [PubMed] [Google Scholar]
  37. von Wedel R. J., Carlson S. S., Kelly R. B. Transfer of synaptic vesicle antigens to the presynaptic plasma membrane during exocytosis. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1014–1018. doi: 10.1073/pnas.78.2.1014. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES