Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1995 Mar 1;128(5):905–912. doi: 10.1083/jcb.128.5.905

Dephosphorylated synapsin I anchors synaptic vesicles to actin cytoskeleton: an analysis by videomicroscopy

PMCID: PMC2120389  PMID: 7876313

Abstract

Synapsin I is a synaptic vesicle-associated protein which inhibits neurotransmitter release, an effect which is abolished upon its phosphorylation by Ca2+/calmodulin-dependent protein kinase II (CaM kinase II). Based on indirect evidence, it was suggested that this effect on neurotransmitter release may be achieved by the reversible anchoring of synaptic vesicles to the actin cytoskeleton of the nerve terminal. Using video-enhanced microscopy, we have now obtained experimental evidence in support of this model: the presence of dephosphorylated synapsin I is necessary for synaptic vesicles to bind actin; synapsin I is able to promote actin polymerization and bundling of actin filaments in the presence of synaptic vesicles; the ability to cross-link synaptic vesicles and actin is specific for synapsin I and is not shared by other basic proteins; the cross-linking between synaptic vesicles and actin is specific for the membrane of synaptic vesicles and does not reflect either a non-specific binding of membranes to the highly surface active synapsin I molecule or trapping of vesicles within the thick bundles of actin filaments; the formation of the ternary complex is virtually abolished when synapsin I is phosphorylated by CaM kinase II. The data indicate that synapsin I markedly affects synaptic vesicle traffic and cytoskeleton assembly in the nerve terminal and provide a molecular basis for the ability of synapsin I to regulate the availability of synaptic vesicles for exocytosis and thereby the efficiency of neurotransmitter release.

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. Arpin M., Friederich E., Algrain M., Vernel F., Louvard D. Functional differences between L- and T-plastin isoforms. J Cell Biol. 1994 Dec;127(6 Pt 2):1995–2008. doi: 10.1083/jcb.127.6.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benfenati F., Greengard P., Brunner J., Bähler M. Electrostatic and hydrophobic interactions of synapsin I and synapsin I fragments with phospholipid bilayers. J Cell Biol. 1989 May;108(5):1851–1862. doi: 10.1083/jcb.108.5.1851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benfenati F., Neyroz P., Bähler M., Masotti L., Greengard P. Time-resolved fluorescence study of the neuron-specific phosphoprotein synapsin I. Evidence for phosphorylation-dependent conformational changes. J Biol Chem. 1990 Jul 25;265(21):12584–12595. [PubMed] [Google Scholar]
  4. Benfenati F., Valtorta F., Chieregatti E., Greengard P. Interaction of free and synaptic vesicle-bound synapsin I with F-actin. Neuron. 1992 Feb;8(2):377–386. doi: 10.1016/0896-6273(92)90303-u. [DOI] [PubMed] [Google Scholar]
  5. Benfenati F., Valtorta F., Greengard P. Computer modeling of synapsin I binding to synaptic vesicles and F-actin: implications for regulation of neurotransmitter release. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):575–579. doi: 10.1073/pnas.88.2.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Benfenati F., Valtorta F., Rubenstein J. L., Gorelick F. S., Greengard P., Czernik A. J. Synaptic vesicle-associated Ca2+/calmodulin-dependent protein kinase II is a binding protein for synapsin I. Nature. 1992 Oct 1;359(6394):417–420. doi: 10.1038/359417a0. [DOI] [PubMed] [Google Scholar]
  7. Bähler M., Benfenati F., Valtorta F., Czernik A. J., Greengard P. Characterization of synapsin I fragments produced by cysteine-specific cleavage: a study of their interactions with F-actin. J Cell Biol. 1989 May;108(5):1841–1849. doi: 10.1083/jcb.108.5.1841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bähler M., Greengard P. Synapsin I bundles F-actin in a phosphorylation-dependent manner. Nature. 1987 Apr 16;326(6114):704–707. doi: 10.1038/326704a0. [DOI] [PubMed] [Google Scholar]
  9. Ceccaldi P. E., Benfenati F., Chieregatti E., Greengard P., Valtorta F. Rapid binding of synapsin I to F- and G-actin. A study using fluorescence resonance energy transfer. FEBS Lett. 1993 Aug 30;329(3):301–305. doi: 10.1016/0014-5793(93)80242-m. [DOI] [PubMed] [Google Scholar]
  10. De Camilli P., Benfenati F., Valtorta F., Greengard P. The synapsins. Annu Rev Cell Biol. 1990;6:433–460. doi: 10.1146/annurev.cb.06.110190.002245. [DOI] [PubMed] [Google Scholar]
  11. Fechheimer M., Zigmond S. H. Focusing on unpolymerized actin. J Cell Biol. 1993 Oct;123(1):1–5. doi: 10.1083/jcb.123.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fesce R., Benfenati F., Greengard P., Valtorta F. Effects of the neuronal phosphoprotein synapsin I on actin polymerization. II. Analytical interpretation of kinetic curves. J Biol Chem. 1992 Jun 5;267(16):11289–11299. [PubMed] [Google Scholar]
  13. Goldstein D., Djeu J., Latter G., Burbeck S., Leavitt J. Abundant synthesis of the transformation-induced protein of neoplastic human fibroblasts, plastin, in normal lymphocytes. Cancer Res. 1985 Nov;45(11 Pt 2):5643–5647. [PubMed] [Google Scholar]
  14. Greengard P., Valtorta F., Czernik A. J., Benfenati F. Synaptic vesicle phosphoproteins and regulation of synaptic function. Science. 1993 Feb 5;259(5096):780–785. doi: 10.1126/science.8430330. [DOI] [PubMed] [Google Scholar]
  15. Grohovaz F., Zacchetti D., Clementi E., Lorenzon P., Meldolesi J., Fumagalli G. [Ca2+]i imaging in PC12 cells: multiple response patterns to receptor activation reveal new aspects of transmembrane signaling. J Cell Biol. 1991 Jun;113(6):1341–1350. doi: 10.1083/jcb.113.6.1341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hackett J. T., Cochran S. L., Greenfield L. J., Jr, Brosius D. C., Ueda T. Synapsin I injected presynaptically into goldfish mauthner axons reduces quantal synaptic transmission. J Neurophysiol. 1990 Apr;63(4):701–706. doi: 10.1152/jn.1990.63.4.701. [DOI] [PubMed] [Google Scholar]
  17. Hirokawa N., Sobue K., Kanda K., Harada A., Yorifuji H. The cytoskeletal architecture of the presynaptic terminal and molecular structure of synapsin 1. J Cell Biol. 1989 Jan;108(1):111–126. doi: 10.1083/jcb.108.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Huttner W. B., Schiebler W., Greengard P., De Camilli P. Synapsin I (protein I), a nerve terminal-specific phosphoprotein. III. Its association with synaptic vesicles studied in a highly purified synaptic vesicle preparation. J Cell Biol. 1983 May;96(5):1374–1388. doi: 10.1083/jcb.96.5.1374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jessell T. M., Kandel E. R. Synaptic transmission: a bidirectional and self-modifiable form of cell-cell communication. Cell. 1993 Jan;72 (Suppl):1–30. doi: 10.1016/s0092-8674(05)80025-x. [DOI] [PubMed] [Google Scholar]
  20. Jovin T. M., Arndt-Jovin D. J. Luminescence digital imaging microscopy. Annu Rev Biophys Biophys Chem. 1989;18:271–308. doi: 10.1146/annurev.bb.18.060189.001415. [DOI] [PubMed] [Google Scholar]
  21. Kelly R. B. Storage and release of neurotransmitters. Cell. 1993 Jan;72 (Suppl):43–53. doi: 10.1016/s0092-8674(05)80027-3. [DOI] [PubMed] [Google Scholar]
  22. Landis D. M., Hall A. K., Weinstein L. A., Reese T. S. The organization of cytoplasm at the presynaptic active zone of a central nervous system synapse. Neuron. 1988 May;1(3):201–209. doi: 10.1016/0896-6273(88)90140-7. [DOI] [PubMed] [Google Scholar]
  23. Llinás R., Gruner J. A., Sugimori M., McGuinness T. L., Greengard P. Regulation by synapsin I and Ca(2+)-calmodulin-dependent protein kinase II of the transmitter release in squid giant synapse. J Physiol. 1991 May;436:257–282. doi: 10.1113/jphysiol.1991.sp018549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Llinás R., McGuinness T. L., Leonard C. S., Sugimori M., Greengard P. Intraterminal injection of synapsin I or calcium/calmodulin-dependent protein kinase II alters neurotransmitter release at the squid giant synapse. Proc Natl Acad Sci U S A. 1985 May;82(9):3035–3039. doi: 10.1073/pnas.82.9.3035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. MacLean-Fletcher S., Pollard T. D. Mechanism of action of cytochalasin B on actin. Cell. 1980 Jun;20(2):329–341. doi: 10.1016/0092-8674(80)90619-4. [DOI] [PubMed] [Google Scholar]
  26. McGuinness T. L., Lai Y., Greengard P. Ca2+/calmodulin-dependent protein kinase II. Isozymic forms from rat forebrain and cerebellum. J Biol Chem. 1985 Feb 10;260(3):1696–1704. [PubMed] [Google Scholar]
  27. Nichols R. A., Chilcote T. J., Czernik A. J., Greengard P. Synapsin I regulates glutamate release from rat brain synaptosomes. J Neurochem. 1992 Feb;58(2):783–785. doi: 10.1111/j.1471-4159.1992.tb09788.x. [DOI] [PubMed] [Google Scholar]
  28. Nichols R. A., Sihra T. S., Czernik A. J., Nairn A. C., Greengard P. Calcium/calmodulin-dependent protein kinase II increases glutamate and noradrenaline release from synaptosomes. Nature. 1990 Feb 15;343(6259):647–651. doi: 10.1038/343647a0. [DOI] [PubMed] [Google Scholar]
  29. Petrucci T. C., Morrow J. S. Synapsin I: an actin-bundling protein under phosphorylation control. J Cell Biol. 1987 Sep;105(3):1355–1363. doi: 10.1083/jcb.105.3.1355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schiebler W., Jahn R., Doucet J. P., Rothlein J., Greengard P. Characterization of synapsin I binding to small synaptic vesicles. J Biol Chem. 1986 Jun 25;261(18):8383–8390. [PubMed] [Google Scholar]
  31. Silva A. J., Stevens C. F., Tonegawa S., Wang Y. Deficient hippocampal long-term potentiation in alpha-calcium-calmodulin kinase II mutant mice. Science. 1992 Jul 10;257(5067):201–206. doi: 10.1126/science.1378648. [DOI] [PubMed] [Google Scholar]
  32. Spudich J. A., Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed] [Google Scholar]
  33. 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]
  34. Valtorta F., Benfenati F., Greengard P. Structure and function of the synapsins. J Biol Chem. 1992 Apr 15;267(11):7195–7198. [PubMed] [Google Scholar]
  35. Valtorta F., Benfenati F. Membrane trafficking in nerve terminals. Adv Pharmacol. 1995;32:505–557. doi: 10.1016/s1054-3589(08)61021-2. [DOI] [PubMed] [Google Scholar]
  36. Valtorta F., Greengard P., Fesce R., Chieregatti E., Benfenati F. Effects of the neuronal phosphoprotein synapsin I on actin polymerization. I. Evidence for a phosphorylation-dependent nucleating effect. J Biol Chem. 1992 Jun 5;267(16):11281–11288. [PubMed] [Google Scholar]
  37. Zucker R. S. Short-term synaptic plasticity. Annu Rev Neurosci. 1989;12:13–31. doi: 10.1146/annurev.ne.12.030189.000305. [DOI] [PubMed] [Google Scholar]

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

RESOURCES