Characterization of synaptic vesicles and related neuronal features in nerve growth factor and ras oncogene differentiated PC12 cells

J. H. Tao-Cheng, A. Dosemeci, Joseph Bressler, M. W. Brightman, D. L. Simpson

Research output: Contribution to journalArticle

Abstract

PC12 cells can differentiate into neuron-like cells after treatment with either nerve growth factor (NGF) or transduction with a retrovirus which expresses the K-ras oncogene. The concomitant treatment of NGF plus ras differentiates PC12 cells further than either agent alone with respect to neurite outgrowth, acetylcholinesterase levels, and most strikingly, the number of synaptic vesicle (SV) clusters. These SV clusters in PC12 cell neurites closely resemble those in the presynaptic terminals of neurons. Such SV clusters have not been described in cell lines previously. The SV clusters from all three differentiated groups (NGF, ras, and NGF plus ras) were similar in size, shape, and configuration, except that the ones in the doubly treated group occur in higher frequency and have more vesicles. The synaptic nature of these vesicle clusters was demonstrated by their regulated depletion after potassium stimulation. Furthermore, these vesicle clusters stained positively for two SV-associated proteins, synapsin I and synaptophysin, by EM immunocytochemistry (ICC). Such SV clusters in a cell line are very useful for characterizing the regulated release of SVs and the distribution of SV-related antigens in intact cells. Analysis by SDS-gel electrophoresis and immunoblotting indicated that synapsin I levels are higher in all three differentiated groups compared to untreated cells; whereas synaptophysin levels are lower in cells exposed to NGF alone or with NGF and ras double treatment. Possible convergence and/or divergence on the mechanisms of NGF and ras differentiation in PC12 cells are discussed.

Original languageEnglish (US)
Pages (from-to)323-334
Number of pages12
JournalJournal of Neuroscience Research
Volume42
Issue number3
DOIs
StatePublished - 1995

Fingerprint

ras Genes
Synaptic Vesicles
PC12 Cells
Nerve Growth Factor
Synapsins
Synaptophysin
Neurons
Cell Line
Presynaptic Terminals
Neurites
Retroviridae
Acetylcholinesterase
Immunoblotting
Electrophoresis
Potassium
Therapeutics
Gels
Immunohistochemistry
Antigens

Keywords

  • acetylcholinesterase
  • EM immunocytochemistry
  • potassium stimulated release
  • synapsin I
  • synaptophysin

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Characterization of synaptic vesicles and related neuronal features in nerve growth factor and ras oncogene differentiated PC12 cells. / Tao-Cheng, J. H.; Dosemeci, A.; Bressler, Joseph; Brightman, M. W.; Simpson, D. L.

In: Journal of Neuroscience Research, Vol. 42, No. 3, 1995, p. 323-334.

Research output: Contribution to journalArticle

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