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. 1983 Dec;80(23):7342–7346. doi: 10.1073/pnas.80.23.7342

A synaptic vesicle antigen is restricted to the junctional region of the presynaptic plasma membrane.

K M Buckley, E S Schweitzer, G P Miljanich, L Clift-O'Grady, P D Kushner, L F Reichardt, R B Kelly
PMCID: PMC390051  PMID: 6359167

Abstract

The plasma membrane of electric organ nerve terminals has two domains that can be distinguished by monoclonal antibodies. A library of 111 mouse monoclonal antibodies raised to nerve terminals from Torpedo californica contains 4 antibodies that bind specifically to the outside of intact synaptosomes. The distribution of the binding sites of these monoclonal antibodies on the outside of intact nerve terminals was examined by immunofluorescence and immunoelectron microscopy. The binding sites of 3 (tor23, 25, and 132) are distributed uniformly over nerve trunks and fine terminal branches. The binding site of the fourth (tor70) is restricted to synaptic junctional regions. This antibody, but not the other 3, recognizes a major component of synaptic vesicles, a proteoglycan associated with the inner surface of the vesicle membrane. The difference in the pattern of binding of these monoclonal antibodies suggests that the region of the plasma membrane containing active zones is antigenically distinguishable from other nerve terminal plasma membrane. We suggest that the antigen recognized by tor70 is externalized by exocytosis of synaptic vesicles while other plasma antigens take a different route to the surface. The unexpected observation that the vesicle antigen remains on the surface after exocytosis and is prevented from diffusion from the synaptic junctional region would be consistent with an interaction between the vesicle proteoglycan and elements of the synaptic cleft.

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Selected References

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

  1. Ash J. F., Louvard D., Singer S. J. Antibody-induced linkages of plasma membrane proteins to intracellular actomyosin-containing filaments in cultured fibroblasts. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5584–5588. doi: 10.1073/pnas.74.12.5584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Branton D., Cohen C. M., Tyler J. Interaction of cytoskeletal proteins on the human erythrocyte membrane. Cell. 1981 Apr;24(1):24–32. doi: 10.1016/0092-8674(81)90497-9. [DOI] [PubMed] [Google Scholar]
  4. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  5. Carlson S. S., Kelly R. B. A highly antigenic proteoglycan-like component of cholinergic synaptic vesicles. J Biol Chem. 1983 Sep 25;258(18):11082–11091. [PubMed] [Google Scholar]
  6. Carlson S. S., Kelly R. B. An antiserum specific for cholinergic synaptic vesicles from electric organ. J Cell Biol. 1980 Oct;87(1):98–103. doi: 10.1083/jcb.87.1.98. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Carlson S. S., Wagner J. A., Kelly R. B. Purification of synaptic vesicles from elasmobranch electric organ and the use of biophysical criteria to demonstrate purity. Biochemistry. 1978 Apr 4;17(7):1188–1199. doi: 10.1021/bi00600a009. [DOI] [PubMed] [Google Scholar]
  8. Ceccarelli B., Grohovaz F., Hurlbut W. P. Freeze-fracture studies of frog neuromuscular junctions during intense release of neurotransmitter. II. Effects of electrical stimulation and high potassium. J Cell Biol. 1979 Apr;81(1):178–192. doi: 10.1083/jcb.81.1.178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Flanagan J., Koch G. L. Cross-linked surface Ig attaches to actin. Nature. 1978 May 25;273(5660):278–281. doi: 10.1038/273278a0. [DOI] [PubMed] [Google Scholar]
  10. Gumbiner B., Kelly R. B. Two distinct intracellular pathways transport secretory and membrane glycoproteins to the surface of pituitary tumor cells. Cell. 1982 Jan;28(1):51–59. doi: 10.1016/0092-8674(82)90374-9. [DOI] [PubMed] [Google Scholar]
  11. Hay E. D. Extracellular matrix. J Cell Biol. 1981 Dec;91(3 Pt 2):205s–223s. doi: 10.1083/jcb.91.3.205s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Heuser J. E., Reese T. S., Dennis M. J., Jan Y., Jan L., Evans L. Synaptic vesicle exocytosis captured by quick freezing and correlated with quantal transmitter release. J Cell Biol. 1979 May;81(2):275–300. doi: 10.1083/jcb.81.2.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Heuser J. E., Salpeter S. R. Organization of acetylcholine receptors in quick-frozen, deep-etched, and rotary-replicated Torpedo postsynaptic membrane. J Cell Biol. 1979 Jul;82(1):150–173. doi: 10.1083/jcb.82.1.150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hooper J. E., Carlson S. S., Kelly R. B. Antibodies to synaptic vesicles purified from Narcine electric organ bind a subclass of mammalian nerve terminals. J Cell Biol. 1980 Oct;87(1):104–113. doi: 10.1083/jcb.87.1.104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hooper J. E., Deutsch J. W., Miljanich G. P., Brasier A. R., Kelly R. B. Nerve terminal components from normal and denervated Narcine electric organ. J Physiol (Paris) 1982;78(4):443–453. [PubMed] [Google Scholar]
  16. Labat-Robert J., Saitoh T., Godeau G., Robert L., Changeux J. P. Distribution of macromolecules from the intercellular matrix in the electroplaque of Electrophorus electricus. FEBS Lett. 1980 Nov 3;120(2):259–263. doi: 10.1016/0014-5793(80)80311-5. [DOI] [PubMed] [Google Scholar]
  17. Lingg G., Fischer-Colbrie R., Schmidt W., Winkler H. Exposure of an antigen of chromaffin granules on cell surface during exocytosis. Nature. 1983 Feb 17;301(5901):610–611. doi: 10.1038/301610a0. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
  20. Miljanich G. P., Brasier A. R., Kelly R. B. Partial purification of presynaptic plasma membrane by immunoadsorption. J Cell Biol. 1982 Jul;94(1):88–96. doi: 10.1083/jcb.94.1.88. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Suszkiw J. B. Kinetics of acetylcholine recovery in Torpedo electromotor synapses depleted of synaptic vesicles. Neuroscience. 1980;5(7):1341–1349. doi: 10.1016/0306-4522(80)90206-7. [DOI] [PubMed] [Google Scholar]
  22. 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]

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