Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1983 May 1;96(5):1337–1354. doi: 10.1083/jcb.96.5.1337

Synapsin I (protein I), a nerve terminal-specific phosphoprotein. I. Its general distribution in synapses of the central and peripheral nervous system demonstrated by immunofluorescence in frozen and plastic sections

PMCID: PMC2112636  PMID: 6404910

Abstract

Synapsin I (formerly referred to as protein I) is the collective name for two almost identical phosphoproteins, synapsin Ia and synapsin Ib (protein Ia and protein Ib), present in the nervous system. Synapsin I has previously been shown by immunoperoxidase studies (De Camilli, P., T. Ueda, F. E. Bloom, E. Battenberg, and P. Greengard, 1979, Proc. Natl. Acad. Sci. USA, 76:5977-5981; Bloom, F. E., T. Ueda, E. Battenberg, and P. Greengard, 1979, Proc. Natl. Acad. Sci. USA 76:5982- 5986) to be a neuron-specific protein, present in both the central and peripheral nervous systems and concentrated in the synaptic region of nerve cells. In those preliminary studies, the occurrence of synapsin I could be demonstrated in only a portion of synapses. We have now carried out a detailed examination of the distribution of synapsin I immunoreactivity in the central and peripheral nervous systems. In this study we have attempted to maximize the level of resolution of immunohistochemical light microscopy images in order to estimate the proportion of immunoreactive synapses and to establish their precise distribution. Optimal results were obtained by the use of immunofluorescence in semithin sections (approximately 1 micron) prepared from Epon-embedded nonosmicated tissues after the Epon had been removed. Our results confirm the previous observations on the specific localization of synapsin I in nerve cells and synapses. In addition, the results strongly suggest that, with a few possible exceptions involving highly specialized neurons, all synapses contain synapsin I. Finally, immunocytochemical experiments indicate that synapsin I appearance in the various regions of the developing nervous system correlates topographically and temporally with the appearance of synapses. In two accompanying papers (De Camilli, P., S. M. Harris, Jr., W. B. Huttner, and P. Greengard, and Huttner, W. B., W. Schiebler, P. Greengard, and P. De Camilli, 1983, J. Cell Biol. 96:1355-1373 and 1374-1388, respectively), evidence is presented that synapsin I is specifically associated with synaptic vesicles in nerve endings.

Full Text

The Full Text of this article is available as a PDF (10.8 MB).

Selected References

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

  1. Baskin D. G., Erlandsen S. L., Parsons J. A. Influence of hydrogen peroxide or alcoholic sodium hydroxide on the immunocytochemical detection of growth hormone and prolactin after osmium fixation. J Histochem Cytochem. 1979 Sep;27(9):1290–1292. doi: 10.1177/27.9.383831. [DOI] [PubMed] [Google Scholar]
  2. Bendayan M., Roth J., Perrelet A., Orci L. Quantitative immunocytochemical localization of pancreatic secretory proteins in subcellular compartments of the rat acinar cell. J Histochem Cytochem. 1980 Feb;28(2):149–160. doi: 10.1177/28.2.7354212. [DOI] [PubMed] [Google Scholar]
  3. Bloom F. E., Ueda T., Battenberg E., Greengard P. Immunocytochemical localization, in synapses, of protein I, an endogenous substrate for protein kinases in mammalian brain. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5982–5986. doi: 10.1073/pnas.76.11.5982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Conradi S. Ultrastructure and distribution of neuronal and glial elements on the motoneuron surface in the lumbosacral spinal cord of the adult cat. Acta Physiol Scand Suppl. 1969;332:5–48. [PubMed] [Google Scholar]
  5. De Camilli P., Harris S. M., Jr, Huttner W. B., Greengard P. Synapsin I (Protein I), a nerve terminal-specific phosphoprotein. II. Its specific association with synaptic vesicles demonstrated by immunocytochemistry in agarose-embedded synaptosomes. J Cell Biol. 1983 May;96(5):1355–1373. doi: 10.1083/jcb.96.5.1355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. De Camilli P., Ueda T., Bloom F. E., Battenberg E., Greengard P. Widespread distribution of protein I in the central and peripheral nervous systems. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5977–5981. doi: 10.1073/pnas.76.11.5977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dolphin A. C., Greengard P. Serotonin stimulates phosphorylation of protein I in the facial motor nucleus of rat brain. Nature. 1981 Jan 1;289(5793):76–79. doi: 10.1038/289076a0. [DOI] [PubMed] [Google Scholar]
  8. Erlandsen S. L., Parsons J. A., Rodning C. B. Technical parameters of immunostaining of osmicated tissue in epoxy sections. J Histochem Cytochem. 1979 Sep;27(9):1286–1289. doi: 10.1177/27.9.90080. [DOI] [PubMed] [Google Scholar]
  9. Forn J., Greengard P. Depolarizing agents and cyclic nucleotides regulate the phosphorylation of specific neuronal proteins in rat cerebral cortex slices. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5195–5199. doi: 10.1073/pnas.75.10.5195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fried G., Nestler E. J., De Camilli P., Stjärne L., Olson L., Lundberg J. M., Hökfelt T., Ouimet C. C., Greengard P. Cellular and subcellular localization of protein I in the peripheral nervous system. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2717–2721. doi: 10.1073/pnas.79.8.2717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gerace L., Blum A., Blobel G. Immunocytochemical localization of the major polypeptides of the nuclear pore complex-lamina fraction. Interphase and mitotic distribution. J Cell Biol. 1978 Nov;79(2 Pt 1):546–566. doi: 10.1083/jcb.79.2.546. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Goelz S. E., Nestler E. J., Chehrazi B., Greengard P. Distribution of protein I in mammalian brain as determined by a detergent-based radioimmunoassay. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2130–2134. doi: 10.1073/pnas.78.4.2130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Huang C. K., Browning M. D., Greengard P. Purification and characterization of protein IIIb, a mammalian brain phosphoprotein. J Biol Chem. 1982 Jun 10;257(11):6524–6528. [PubMed] [Google Scholar]
  14. Huttner W. B., DeGennaro L. J., Greengard P. Differential phosphorylation of multiple sites in purified protein I by cyclic AMP-dependent and calcium-dependent protein kinases. J Biol Chem. 1981 Feb 10;256(3):1482–1488. [PubMed] [Google Scholar]
  15. Huttner W. B., Greengard P. Multiple phosphorylation sites in protein I and their differential regulation by cyclic AMP and calcium. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5402–5406. doi: 10.1073/pnas.76.10.5402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Johnson E. M., Ueda T., Maeno H., Greengard P. Adenosine 3',5-monophosphate-dependent phosphorylation of a specific protein in synaptic membrane fractions from rat cerebrum. J Biol Chem. 1972 Sep 10;247(17):5650–5652. [PubMed] [Google Scholar]
  18. Kennedy M. B., Greengard P. Two calcium/calmodulin-dependent protein kinases, which are highly concentrated in brain, phosphorylate protein I at distinct sites. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1293–1297. doi: 10.1073/pnas.78.2.1293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Krueger B. K., Forn J., Greengard P. Depolarization-induced phosphorylation of specific proteins, mediated by calcium ion influx, in rat brain synaptosomes. J Biol Chem. 1977 Apr 25;252(8):2764–2773. [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Matthew W. D., Tsavaler L., Reichardt L. F. Identification of a synaptic vesicle-specific membrane protein with a wide distribution in neuronal and neurosecretory tissue. J Cell Biol. 1981 Oct;91(1):257–269. doi: 10.1083/jcb.91.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Maxwell M. H. Two rapid and simple methods used for the removal of resins from 1.0 micron thick epoxy sections. J Microsc. 1978 Mar;112(2):253–255. doi: 10.1111/j.1365-2818.1978.tb01174.x. [DOI] [PubMed] [Google Scholar]
  24. Moriarty G. C. Adenohypophysis: ultrastructural cytochemistry. A review. J Histochem Cytochem. 1973 Oct;21(10):855–894. doi: 10.1177/21.10.855. [DOI] [PubMed] [Google Scholar]
  25. Nestler E. J., Greengard P. Dopamine and depolarizing agents regulate the state of phosphorylation of protein I in the mammalian superior cervical sympathetic ganglion. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7479–7483. doi: 10.1073/pnas.77.12.7479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
  27. Pelletier G., Puviani R., Bosler O., Descarries L. Immunocytochemical detection of peptides in osmicated and plastic-embedded tissue. An electron microscopic study. J Histochem Cytochem. 1981 Jun;29(6):759–764. doi: 10.1177/29.6.7019313. [DOI] [PubMed] [Google Scholar]
  28. Pérez de la Mora M., Possani L. D., Tapia R., Teran L., Palacios R., Fuxe K., Hökfelt T., Ljungdahl A. Demonstration of central gamma-aminobutyrate-containing nerve terminals by means of antibodies against glutamate decarboxylase. Neuroscience. 1981;6(5):875–895. doi: 10.1016/0306-4522(81)90169-x. [DOI] [PubMed] [Google Scholar]
  29. Rinvik E., Grofová I. Observations on the fine structure of the substantia nigra in the cat. Exp Brain Res. 1970;11(3):229–248. doi: 10.1007/BF01474384. [DOI] [PubMed] [Google Scholar]
  30. Schaeffer S. F., Raviola E. Membrane recycling in the cone cell endings of the turtle retina. J Cell Biol. 1978 Dec;79(3):802–825. doi: 10.1083/jcb.79.3.802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sieghart W., Forn J., Greengard P. Ca2+ and cyclic AMP regulate phosphorylation of same two membrane-associated proteins specific to nerve tissue. Proc Natl Acad Sci U S A. 1979 May;76(5):2475–2479. doi: 10.1073/pnas.76.5.2475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Swanson L. W., Wyss J. M., Cowan W. M. An autoradiographic study of the organization of intrahippocampal association pathways in the rat. J Comp Neurol. 1978 Oct 15;181(4):681–715. doi: 10.1002/cne.901810402. [DOI] [PubMed] [Google Scholar]
  33. Thureson-Klein A., Klein R. L., Johansson O. Catecholamine-rich cells and varicosities in bovine splenic nerve, vesicle contents and evidence for exocytosis. J Neurobiol. 1979 May;10(3):309–324. doi: 10.1002/neu.480100309. [DOI] [PubMed] [Google Scholar]
  34. Ueda T., Greengard P. Adenosine 3':5'-monophosphate-regulated phosphoprotein system of neuronal membranes. I. Solubilization, purification, and some properties of an endogenous phosphoprotein. J Biol Chem. 1977 Jul 25;252(14):5155–5163. [PubMed] [Google Scholar]
  35. Ueda T., Greengard P., Berzins K., Cohen R. S., Blomberg F., Grab D. J., Siekevitz P. Subcellular distribution in cerebral cortex of two proteins phosphorylated by a cAMP-dependent protein kinase. J Cell Biol. 1979 Nov;83(2 Pt 1):308–319. doi: 10.1083/jcb.83.2.308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Walaas I., Fonnum F. The distribution and origin of glutamate decarboxylase and choline acetyltransferase in ventral pallidum and other basal forebrain regions. Brain Res. 1979 Nov 16;177(2):325–336. doi: 10.1016/0006-8993(79)90783-2. [DOI] [PubMed] [Google Scholar]
  37. Weeke B. A manual of quantitative immunoelectrophoresis. Methods and applications. 1. General remarks on principles, equipment, reagents and procedures. Scand J Immunol Suppl. 1973;1:15–35. doi: 10.1111/j.1365-3083.1973.tb03776.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES