Expression and localization of Lewisx glycolipids and GD1a ganglioside in human glioma cells

Toshio Ariga, Shama Bhat, Takashi Kanda, Masanaga Yamawaki, Tadashi Tai, Yasunori Kushi, Takeshi Kasama, Shizuo Handa, Robert K Yu

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

We analysed the glycolipid composition of glioma cells (N-370 FG cells), which are derived from a culture of transformed human fetal glial cells. The neutral and acidic glycolipid fractions were isolated by column chromatography on DEAE-Sephadex and analysed by high-performance thin-layer chromatography (HPTLC). The neutral glycolipid fraction contained 1.6 μg of lipid-bound glucose/galactose per mg protein and consisted of GlcCer (11.4% of total neutral glycolipids), GalCer (21.5%), LacCer (21.4%), Gb4 (21.1%), and three unknown neutral glycolipids (23%). These unknown glycolipids were characterized as Lewisx (fucosylneolactonorpentaosyl ceramide; Lex), difucosylneolactonorhexaosyl ceramide (dimeric Lex), and neolactonorhexaosyl ceramide (nLc6) by an HPTLC-overlay method for glycolipids using specific mouse anti-glycolipid antibodies against glycolipid and/or liquid-secondary ion (LSI) mass spectrometry. The ganglioside fraction contained 0.6 μg of lipid-bound sialic acid per mg protein with GD1a as the predominant ganglioside species (83% of the total gangliosides) and GM3, GM2, and GM1 as minor components. Trace amounts of sialyl-Lex and the complex type of sialyl-Lex derivatives were also present. Immunocytochemical studies revealed that GD1a and GalCer were primarily localized on the surface of cell bodies. Interestingly, Lex glycolipids and sialyl-Lex were localized not only on the cell bodies but also on short cell processes. Especially, sialyl-Lex glycolipid was located on the tip of fine cellular processes. The unique localization of the Lex glycolipids suggests that they may be involved in cellular differentiation and initiation of cellular growth in this cell line.

Original languageEnglish (US)
Pages (from-to)135-145
Number of pages11
JournalGlycoconjugate Journal
Volume13
Issue number2
DOIs
StatePublished - Jan 1 1996
Externally publishedYes

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Glycolipids
Glioma
Ceramides
Thin layer chromatography
Gangliosides
Cells
Thin Layer Chromatography
GD1a ganglioside
G(M2) Ganglioside
G(M3) Ganglioside
Secondary Ion Mass Spectrometry
G(M1) Ganglioside
Lipids
DEAE-Dextran
Column chromatography
N-Acetylneuraminic Acid
Secondary ion mass spectrometry
Galactose
Neuroglia
Chromatography

Keywords

  • Gangliosides
  • Glycolipids
  • Human glioma cells
  • Lewis
  • N-370 FG cells
  • Neutral glycolipids

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Expression and localization of Lewisx glycolipids and GD1a ganglioside in human glioma cells. / Ariga, Toshio; Bhat, Shama; Kanda, Takashi; Yamawaki, Masanaga; Tai, Tadashi; Kushi, Yasunori; Kasama, Takeshi; Handa, Shizuo; Yu, Robert K.

In: Glycoconjugate Journal, Vol. 13, No. 2, 01.01.1996, p. 135-145.

Research output: Contribution to journalArticle

Ariga, T, Bhat, S, Kanda, T, Yamawaki, M, Tai, T, Kushi, Y, Kasama, T, Handa, S & Yu, RK 1996, 'Expression and localization of Lewisx glycolipids and GD1a ganglioside in human glioma cells', Glycoconjugate Journal, vol. 13, no. 2, pp. 135-145. https://doi.org/10.1007/BF00731487
Ariga, Toshio ; Bhat, Shama ; Kanda, Takashi ; Yamawaki, Masanaga ; Tai, Tadashi ; Kushi, Yasunori ; Kasama, Takeshi ; Handa, Shizuo ; Yu, Robert K. / Expression and localization of Lewisx glycolipids and GD1a ganglioside in human glioma cells. In: Glycoconjugate Journal. 1996 ; Vol. 13, No. 2. pp. 135-145.
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abstract = "We analysed the glycolipid composition of glioma cells (N-370 FG cells), which are derived from a culture of transformed human fetal glial cells. The neutral and acidic glycolipid fractions were isolated by column chromatography on DEAE-Sephadex and analysed by high-performance thin-layer chromatography (HPTLC). The neutral glycolipid fraction contained 1.6 μg of lipid-bound glucose/galactose per mg protein and consisted of GlcCer (11.4{\%} of total neutral glycolipids), GalCer (21.5{\%}), LacCer (21.4{\%}), Gb4 (21.1{\%}), and three unknown neutral glycolipids (23{\%}). These unknown glycolipids were characterized as Lewisx (fucosylneolactonorpentaosyl ceramide; Lex), difucosylneolactonorhexaosyl ceramide (dimeric Lex), and neolactonorhexaosyl ceramide (nLc6) by an HPTLC-overlay method for glycolipids using specific mouse anti-glycolipid antibodies against glycolipid and/or liquid-secondary ion (LSI) mass spectrometry. The ganglioside fraction contained 0.6 μg of lipid-bound sialic acid per mg protein with GD1a as the predominant ganglioside species (83{\%} of the total gangliosides) and GM3, GM2, and GM1 as minor components. Trace amounts of sialyl-Lex and the complex type of sialyl-Lex derivatives were also present. Immunocytochemical studies revealed that GD1a and GalCer were primarily localized on the surface of cell bodies. Interestingly, Lex glycolipids and sialyl-Lex were localized not only on the cell bodies but also on short cell processes. Especially, sialyl-Lex glycolipid was located on the tip of fine cellular processes. The unique localization of the Lex glycolipids suggests that they may be involved in cellular differentiation and initiation of cellular growth in this cell line.",
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