Summary
Cell cultures of the rat cerebellum were immunostained with antibodies to synaptic vesicle antigens, Synapsin I and SV48. Light microscopic immunocytochemistry showed that the initial appearance of demonstrable SV48 and Synapsin I immunoreactivity occurred at different times. Synapsin I immunostaining, unlike SV48 immunostaining, was first seen at 3 daysin vitro as occasional punctate immunofluorescence in neurites, while SV48 immunostaining was first seen at 5 daysin vitro. Both SV48 and Synapsin I punctate immunostaining became frequent at 7 daysin vitro. Double labelling experiments showed coexistence of the above proteins in punctate swellings and growth cones. Using the electron microscope, either SV48 or Synapsin I immunostaining was demonstrated within presynaptic elements in the neuropil. When cultures were incubated with polylysine-coated beads, both types of immunostaining were found in the vesicle containing presynaptic elements formed on the bead surface. It is concluded that Synapsin I and SV48 are (1) co-localized in the same populations of presynaptic elements, (2) co-localized in some growth cones and (3) found in presynaptic elements on beads.
Similar content being viewed by others
References
Bixby, J. L. &Reichardt, L. F. (1985) The expression of synaptic vesicle antigens at neuromuscular junctionsin vitro.Journal of Neuroscience 5, 3070–80.
Bloom, F. E., Ueda, T., Battenberg, E. &Greengard, P. (1979) Immunocytochemical localization, in synapses, of protein I, an endogenous substrate for protein kinase in mammalian brain.Proceedings of the National Academy of Sciences USA 76, 5892–6.
Bunge, M. B. (1973) Fine structure of nerve fibers and growth cones of isolated sympathetic neurons in culture.Journal of Cell Biology 56, 713–35.
Burry, R. W. (1980) Formation of apparent presynaptic elements in response to poly-basic compounds.Brain Research 184, 85–98.
Burry, R. W. (1982) Development of apparent presynaptic elements formed in response to polylysine coated surfaces.Brain Research 247, 1–16.
Burry, R. W. (1983a) Postnatal rat neurons form apparent presynaptic elements on polylysine-coated beadsin vivo.Brain Research 278, 236–9.
Burry, R. W. (1983b) Antimitotic drugs that enhance neuronal survival in olfactory bulb cell cultures.Brain Research 261, 261–75.
Burry, R. W. (1985) Protein synthesis requirement for the formation of synaptic elements.Brain Research 344, 109–19.
Burry, R. W. &Hayes, D. M. (1986) Development and elimination of presynaptic elements on polylysine coated beads implanted in neonatal rat cerebellar.Journal of Neuroscience Research 15, 67–78.
Burry, R. W., Kniss, D. A. &Ho, R. H. (1985) Enhanced survival of apparent presynaptic elements by inhibition of non-neuronal cell proliferation.Brain Research 346, 42–50.
Burry, R. W. &Lasher, R. S. (1978) A quantitative electron microscopic study of synapse formation in dispersed cell cultures in rat cerebellum stained either by Os-UL or by E-PTA.Brain Research 147, 1–15.
De Camilli, P., Cameron, R. &Greengard, P. (1983a) 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.Journal of Cell Biology 96, 1337–54.
De Camilli, P., Harris, S. M., Huttner, W. B. &Greengard, P. (1983b) Synapsin I (protein I), a nerve terminal-specific phosphoprotein. II. Its specific association with synaptic vesicles demonstrated by immunocytochemistry in agarose-embedded synaptosomes.Journal of Cell Biology 96, 1355–73.
De Camilli, P., Ueda, T., Bloom, F. E., Battenberg, E. &Greengard, P. (1979) Widespread distribution of protein I in the central and peripheral nervous system.Proceedings of the National Academy of Sciences USA 76, 5977–81.
Dimpfel, W., Huang, R. T. C. &Habermann, E. (1977) Gangliosides in nervous tissue cultures and binding of125I-labeled tetanus toxin, a neuronal marker.Journal of Neurochemistry 29, 329–34.
Dimpfel, W., Neale, J. H. &Habermann, E. (1975)125I-labelled tetanus toxin as a neuronal marker in tissue cultures derived from embryonic CNS.Naunyn Schniedeberg's Archives of Pharmacology (Berlin) 290, 329–33.
Huttner, W. B., Schiebler, W., Greengard, P. &De Camilli, P. (1983) Synapsin I (protein I), a nerve terminal-specific phosphoprotein. III. Its association with synaptic vesicles studied in a highly purified synaptic vesicle preparation.Journal of Cell Biology 96, 1374–88.
Landis, S. C. (1983) Neuronal growth cones.Annual Review of Physiology 45, 567–80.
Lasher, R. S. (1974) The uptake of (3H) GABA and differentiation of stellate neurons in cultures of dissociated postnatal rat cerebellum.Brain Research 69, 235–54.
Matthew, W. D., Tsavaller, L. &Reichardt, L. F. (1981) Identification of a synaptic vesicle-specific membrane protein with a wide distribution of neuronal and neurosecretory tissue.Journal of Cell Biology 91, 257–69.
Pfenninger, K. H. &Maylie-Pfenninger, M.-F. (1981) Lectin labeling of sprouting neurons. II. Relative movement and appearance of glycoconjugates during plasmalemmal expansion.Journal of Cell Biology 89, 457–559.
Steward, O. &Falk, P. M. (1985) Polyribosomes under developing spine synapses: growth specializations of dendrites at sites of synaptogenesis.Journal of Neuroscience Research 13, 75–88.
Ueda, T. &Greengard, P. (1977) Adenosine 3′,5′-monophosphate-regulated phosphoprotein system of neuronal membranes. I. Solubilization, purification and some properties of an endogeneous phosphoprotein.Journal of Biological Chemistry 252, 5155–63.
Yamada, K. M., Spooner, B. S. &Wessells, N. K. (1971) Ultrastructure and function of growth cones and axons of cultures nerve cells.Journal of Cell Biology 49, 614–35.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Burry, R.W., Raymond, H.H. & Matthew, W.D. Presynaptic elements formed on polylysine-coated beads contain synaptic vesicle antigens. J Neurocytol 15, 409–419 (1986). https://doi.org/10.1007/BF01611725
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01611725