Expression of gangliosides in neuronal development of P19 embryonal carcinoma stem cells

Sean S. Liour, Dmitri Kapitonov, Robert K. Yu

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Gangliosides are constituents of the cell membrane and are known to have important functions in neuronal differentiation. We employed an embryonal carcinoma stem cell line P19 as an in vitro model to investigate the expression of gangliosides during neuronal development. After treatment with retinoic acid, these cells differentiate synchronously into neuron-like cells by a series of well-defined events of development. We examined several aspects of ganglioside metabolism, including the changes of ganglioside pattern, the activities and gene expression of several enzymes at different stages of differentiation, and the distribution of gangliosides in differentiating neurons. Undifferentiated P19 cells express mainly GM3 and GD3. After P19 cells were committed to differentiation, the synthesis of complex gangliosides was elevated more than 20-fold, coinciding with the stage of neurite outgrowth. During the maturation of differentiated cells, the expression of c-series gangliosides was downregulated concomitantly with upregulation of the expression of a- and b-series gangliosides. We also examined the distribution of gangliosides in differentiating neurons by confocal and transmission electron microscopy after cholera toxin B subunit and sialidase treatment. Confocal microscopic studies showed that gangliosides were distributed on the growth cones and exhibited a punctate localization on neurites and soma. Electron microscopic studies indicated that they also are enriched on the plasma membranes of neurites and the filopodia as well as on the lamellipodia of growth cones during the early stage of neurite outgrowth. Our data demonstrate that the expression of gangliosides in P19 cells during RA-induced neuronal differentiation resembles that of the in vivo development of the vertebrate brain, and hence validates it as an in vitro model for investigating the function of gangliosides in neuronal development. (C) 2000 Wiley-Liss, Inc.

Original languageEnglish (US)
Pages (from-to)363-373
Number of pages11
JournalJournal of Neuroscience Research
Volume62
Issue number3
DOIs
StatePublished - Nov 1 2000

Fingerprint

Embryonal Carcinoma Stem Cells
Gangliosides
Growth Cones
Pseudopodia
Neurites
Neurons
Cell Membrane
Cholera Toxin
Neuraminidase
Carisoprodol
Tretinoin
Transmission Electron Microscopy
Vertebrates

Keywords

  • Embryonal carcinoma stem cell
  • Ganglioside
  • Glycosyltransferase
  • Growth cone
  • Neurite outgrowth
  • Neuronal differentiation

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

Cite this

Expression of gangliosides in neuronal development of P19 embryonal carcinoma stem cells. / Liour, Sean S.; Kapitonov, Dmitri; Yu, Robert K.

In: Journal of Neuroscience Research, Vol. 62, No. 3, 01.11.2000, p. 363-373.

Research output: Contribution to journalArticle

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abstract = "Gangliosides are constituents of the cell membrane and are known to have important functions in neuronal differentiation. We employed an embryonal carcinoma stem cell line P19 as an in vitro model to investigate the expression of gangliosides during neuronal development. After treatment with retinoic acid, these cells differentiate synchronously into neuron-like cells by a series of well-defined events of development. We examined several aspects of ganglioside metabolism, including the changes of ganglioside pattern, the activities and gene expression of several enzymes at different stages of differentiation, and the distribution of gangliosides in differentiating neurons. Undifferentiated P19 cells express mainly GM3 and GD3. After P19 cells were committed to differentiation, the synthesis of complex gangliosides was elevated more than 20-fold, coinciding with the stage of neurite outgrowth. During the maturation of differentiated cells, the expression of c-series gangliosides was downregulated concomitantly with upregulation of the expression of a- and b-series gangliosides. We also examined the distribution of gangliosides in differentiating neurons by confocal and transmission electron microscopy after cholera toxin B subunit and sialidase treatment. Confocal microscopic studies showed that gangliosides were distributed on the growth cones and exhibited a punctate localization on neurites and soma. Electron microscopic studies indicated that they also are enriched on the plasma membranes of neurites and the filopodia as well as on the lamellipodia of growth cones during the early stage of neurite outgrowth. Our data demonstrate that the expression of gangliosides in P19 cells during RA-induced neuronal differentiation resembles that of the in vivo development of the vertebrate brain, and hence validates it as an in vitro model for investigating the function of gangliosides in neuronal development. (C) 2000 Wiley-Liss, Inc.",
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