| Project/Area Number |
02680130
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
物質生物化学
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| Research Institution | Shinshu University |
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Principal Investigator |
UEMURA Kei-ichi Associate Professor, Department of Lipid Biochemistry, Shinshu University School of Medicine, 医・心脈管病研, 助教授 (80012756)
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| Co-Investigator(Kenkyū-buntansha) |
TAKETOMI Tamotsu Professor, Department of Lipid Biochemistry, Shinshu University School of Medici, 医・心脈管病研, 教授 (30020704)
HARA Atsushi Assistant Professor, Department of Lipid Biochemistry, Shinshu University School, 医・心脈管病研, 講師 (70126697)
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| Project Period (FY) |
1990 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1992: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1990: ¥700,000 (Direct Cost: ¥700,000)
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| Keywords | Glycosphingolipid / Glycolipid biosynthesis / Sphingolipid / Ganglioside / Neuroblastoma cell / Neurite outgrowth / Cell differentiation / Cell growth / 糖転移酵素 |
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| Research Abstract |
1-Phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), an effective inhibitor of UDP-glucose:ceramide glucosyltransferase, caused inhibition of cell growth in murine neuroblastoma cell lines. Metabolic labeling of glycosphingolipids with [14C]galactose in NS-20Y, Neuro2a, and N1E-115 cells showed reduced incorporation of radioactivity into gangliosides and neutral glycosphingolipids when threo-PDMP was present in the medium. Treatment of NS-20Y cells with threo-PDMP resulted in a time-dependent decrease in mass levels of gangliosides and neutral glycosphingolipids. After 24h in the presence of 50muM threo-PDMP, neutral glycosphingolipid mass was reduced to 32%, where glucosylceramide was the most affected (90% decrease). The ganglioside mass was reduced to 57% of the original content. Neurite outgrowth from neuroblastoma cells in serum-free medium was significantly inhibited by threo-PDMP in a dose-dependent manner. Threo-PDMP also caused retraction of neurites which had been induced to extend in serum-free medium. Pretreatment of cells with GM1 partially restored the ability of NS-20Y cells for neurite outgrowth in the medium containing threo-PDMP. These results suggest a possible role for glycosphingolilids in neurite outgrowth of murine neuroblastoma cells. Ceramide, sphingomyelin and sphingo-sine were found to accumulate in NS-20Y cells after treatment with threo-PDMP. Exogenous sphingosine inhibited neurite outgrowth and caused retraction of neurites. Neurite outgrowth from NS-20Y cells was inhibited by N,N-dimethyl-sphingosine (50% inhibition at -0.1 muM), sphingosine(-0.8 muM),N-hexanoyl-sphingosine (-1 muM), and N-acethylsphingosine(-10muM). A protein kinase inhibitor, H-7, did not affect the neurite outgrowth, suggesting the inhibitory effect of these sphingolipids is protein kinase C-independent.
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