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1996 Fiscal Year Final Research Report Summary

Sphingolipids which regulate the cell functions in neuronal cells.

Research Project

Project/Area Number 06680757
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Neurochemistry/Neuropharmacology
Research InstitutionShinshu University

Principal Investigator

UEMURA Kei-ichi  Shinshu University School of Medicine, Research Center for Aging and Adaptation, Department of Aging Biochemistry, Associate Professor, 医学部・加齢適応研究センター, 助教授 (80012756)

Co-Investigator(Kenkyū-buntansha) TAKETOMI Tamotsu  Shinshu University School of Medicine, Research Center for Aging and Adaptation,, 医学部, 教授 (30020704)
HARA Atsushi  Shinshu University School of Medicine, Research Center for Aging and Adaptation,, 医学部, 講師 (70126697)
Project Period (FY) 1994 – 1996
Keywordssphingolipid / sphingosine / neuroblastoma cell / neurite / glycolipid / protein kinase / calmodulin / cell cycle
Research Abstract

(1) Neurite outgrowth in murine neuroblastoma NS-20Y cells induced by serum deprivation was inhibited by shingosine (1-2muM) as well as by an inhibitor of glucosylceramide synthase (1-phenyl-2-decanoylamino-3-morpholino-3-propanol). The roles of protein kinases and calmodulin in regulating neurite outgrowth were investigated by testing the effect of several inhibitors on the neuritogenesis. W-7 (10muM), calmidazolium (0.3muM) and trifluoperazine (0.1muM), drugs reported to inhibit calmodulin-dependent events, reduced neurite outgrowth. On the other hand, H-7 and H-89 were ineffective, and genistein did not affect the number of cells bearing neurites. Activation of protein kinases, which are blocked by these inhibitors, does not appear to be essential to the extension and maintenance of neurites. KN-62 and KN-93 (inhibitors of Ca^<2+>/calmodulin-dependent protein kinase II) did not inhibit neurite outgrowth. These results suggest that a calmodulin-dependent process, other than the activ … More ation of Ca^<2+>/calmodulin-dependent protein kinase II,is involved in neuritogenesis of murine neuroblastoma NS-20Y cells in serum-free medium. The inhibitory effect of sphingosine on neurite outgrowth is most likely due to calmodulin inhibition.
(2) The effects of a series of sphingosine derivatives and an inhibitor of glucosylceramide synthase, PDMP (1-phenyl-2-decanoylamino-3-morpholino-1-propanol), on the cell cycle progression of murine neuroblastoma NS-20Y cells were investigated by flow cytometry. In the presence of PDMP (50muM,24h) the proportion of cells in the S phase was reduced, whereas that in the G0/G1 phase significantly increased. Incubation of cells with sphingosine, sphinganine, sphingosine-1-phosphate, short-chain ceramides (N-acethyl-or N-hexanoyl-sphingosine) or sphingosyl-phosphocholine did not affect the cell cycle. The DNA peaks characteristic of apoptosis were not observed. These results suggest that the inhibition of the proliferation of NS-20Y cells by PDMP is caused by its effect on cell cycle progression and is more likely due to the reduced levels of glycosphingolipids rather than the increased levels of ceramide or sphingosine. Less

  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] K. Shimada: "Suppression of experimental allergic encephalomyelitis in Lewis rats by administration of gangliosides." Cell, Immunol.154. 231-239 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H. Kurahashi: "Evidence for indirect utilization of glycine for production of N-bases of glycosphingolipids and phospholipids in mouse neuroblastoma cells by using ^5N-labeled glycine" Biochem. Biophys. Res. Commun.210. 44-50 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K. Uemura: "Effect of protein kinase inhibitors and calmodulin inhibitors on neurite outgrowth in murine neuroblastoma NS-20Y cells." J. Biochem.118. 371-375 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A. Hara: "Sulfatide prolongs blood coagulation and bleeding time by forming a complex with fibrinogen." Glycoconjugate J.13. 187-194 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T. Taketomi: "A microwave-mediated saponification of galactosylceramide and galactosylceramide I^3-sulfate and identification of their lyso-compounds by delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry." Biochem. Biophys. Res. Commun.224. 462-467 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T. Taketomi: "Rapid method of preparation of lysoglycosphingolipids and their confirmation by delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry." J. Biochem.120. 573-579 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shimada, K., Koh, C.-S., Uemura, K.and Yanagisawa, H.: "Suppression of experimental allergic encephalomyelitis in Lewis rats by administration of gangliosides." Cell. Immunol.154. 231-239 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ono, K., Hattori, H., Uemura, K., Nakayama, J., Ohta, H.and Katsuyama, T.: "Expression of Forssman antigen in human large intestine" J.Histochem. Cytochem.42. 659-665 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kurahashi, H., Uemura, K., Hara, A.and Taketomi, T.: "Evidence for indirect utilization of glycine for production of N-bases of glycosphingolipids and phospholipids in mouse neuroblastoma cells by using ^<15>N-labeled glycine." Biochem. Biophys. Res. Commun.210. 44-50 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Uemura, K.and Taketomi, T.: "Effects of protein kinase inhibitors and calmodulin inhibitors on neurite outgrowth in murine neuroblastoma NS-20Y cells." J.Biochem.118. 371-375 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ono, K., Oshiro, M., Uemura, K., Ota, H., Matsushita, Y., Ijima, S., Iwase, T., Uchida, M.and Katsuyama, T.: "Erdheim-Chester disease : A case report with immunohistochemical and biochemical examination." Hum. Pathol.27. 91-95 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hara, A., Uemura, K.and Taketomi, T.: "Sulfatide prolongs blood-coagulation and bleeding time by forming a complex with fibrinogen." Glycoconjugate J.13. 187-194 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Taketomi, T., Hara, A., Uemura, K., Kurahashi, H.and Sugiyama, E.: "A microwave-mediated saponification of galactosylceramide and galactosylceramide I^3-sulfate and identification of their lyso-compounds by delayd extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Biochem. Biophys." Res. Commun.224. 462-467 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Taketomi, T., Hara, A., Uemura, K.and Sugiyama, E.: "Rapid method of preparation of lysoglycosphingolipids and their confirmation by delayd extraction matrix-assisted laser desorption ionization time-of flight mass spectrometry." J.Biochem.120. 573-579 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Uemura, K.and Taketomi, T.: "Effects of protein kinase inhibitors and calmodulin inhibitors on neurite outgrowth in murine neuroblastoma NS-20Y cells." J.Neurochem.65. 84 (1995)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1999-03-09  

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