Intracellular signal transduction mechanims of neuronal survival and death

• Project/Area Number: 11670616
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Neurology
• Research Institution: Fukui Medical University
• Principal Investigator: MUTOH Tatsuro Fukui Medical University Hospital Lecturer, 医学部・附属病院, 講師 (60190857)
• Co-Investigator(Kenkyū-buntansha): HAMAGUCHI Michinari Nagoya University School of Medicine Prof., 医学部, 教授 (90135351)
• Project Period (FY): 1999 – 2000
• Project Status: Completed (Fiscal Year 2000)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2000: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 1999: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: Ganglioside / NGF / Trk / GM1 / raft / apoptosis / Signal transduction / Trk / Raft / GM1ガングリオシド / 神経栄養因子 / チロシンキナーゼ / カヴェオラ

Research Abstract

Our previous studies have shown that gangliosideGM1 (GM1), a major constiuent of membrane raft, regulates high affinity nerve growth factor receptor, Trk, which is an essential factor for neuronal differentiation and survival. We constructed mutant human trk cDNA which is lacking possible N-glycosylation site and transfected into PCnnr cells which lacks Trk expression. We have been able to get the stable transfectant which overexpress mutant trk cDNA.This mutant clone did not show any response to NGF in terms of tyrosine autophosphorylation response, neurite outgrowth, and the prevention of serum-deprivation-induced apoptosis. These mutant Trk protein is not located in the lipids raft but rather distributed in higher density fractions of sucrose density gradient ultracentrifugation. To validate the above data, we also examined the effect of cholesterol kilating agents such as β-methyl cyclodextrin and filipin on NGF-induced Trk autophosphorylation response, because these agents has been reported to disrupt raft structure. We did not observe any reponses to NGF in cells pretreated with these agents. These data strongly suggest that GM1 and membrane raft structure are important factors for the normal function of the Trk receptor protein.

Research Products

• [Publications] Mutoh T, et al: "Glucosylceramide synthase inhibitor inhibits the action of nerve growth factor in PC12 cells"J Biol chem. 273. 26001-26007 (1998)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Pitto M,Mutoh T, et al: "Influence of endogenous GM1 ganglioside on Trk B activity in cultured cerebellar granule cells"FEBS Lett. 483. 93-96 (1998)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Mutoh T et al.: "Involvement of tyrosine phosphorylation in HMG-CoA reductase inhibitor-induced cell death in L6 myoblasts"FEBS Lett. 444. 85-89 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Mutoh T, et al: "Role of tyrosine phosphorylation of phospholipase C-γ1 in the signal transduction pathway of HMG-CoA reductase inhitor-"FEBS Lett. 446. 91-94 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Fukumoto S,Mutoh T et al: "GD3 synthase gene expression in PC12 cells resulted in the continuous activation FKA and ERK 1/2, leading to"J Biol Chem. 275. 5832-5838 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Mutoh T et al.: "Unglycosylated TrK protein does not co-localize nor associate with ganglioside GM1 in stable clone.-"Glyconj J. 17. 233-237 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Mutoh T, Kumano T, Nakagawa H, Kuriyama M.: "Involvement of tyrosine phosphorylation in HMG-CoA reductase inhibitor-induced cell death in L6 myoblasts."FEBS Lett. 444. 85-89 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Mutoh T, Kumano T, Nakagawa H, Kuriyama M.: "Role of tyrosine phosphorylation of phospholipase C-γ1 in the signal transduction pathway of HMG-CoA reductase inhibitor-induced cell death in L6 myoblasts."FEBS Lett. 446. 91-94 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Mitsuma N, Yamamoto M, Li M, Ito Y, Mitsuma T, Mutoh T, Takahashi M, Sobue G.: "Expression of GDNF receptor (Ret and GDNFR-α) mRNAs in the spinal cord of patients with amyotrophic lateral sclerosis."Brain Res. 820. 77-85 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Hamano T, Mutoh T, Hirayama M, Kuriyama M.: "MRI finding of benign monomeric amyotrophy of lower limb."J Neurol Sci. 165. 184-187 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Palestini P, Masserini M, Bottiroil G, Brunner J, Mutoh T, Ferraretto AA, Ravasi D, Pitto M.: "Involvement of glycolipid-enriched domains in the transduction mechanism of neurotrophins in cultured neurons."Biosci Rep. 19. 285-295 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Mutoh T, Sobue G, Hamano T, Kuriyama M, Hirayama M, Yamamoto M, Mitsuma T.: "Decreased tyrosine phosphorylation of TrkB receptor in the spinal cords from the patients with amyotrophic lateral sclerosis."Neurochem Res. 25. 239-245 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Hamano T, Mutoh T, Sugie H, Koga H, Kuriyama M.: "Phosphoglycerate kinase deficiency. An adult myopathic form with a novel mutation."Neurology. 54. 1188-1190 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kumano T, Mutoh T, Nakagawa H, Kuriyama M.: "HMG-CoA reductase inhibitor induces a transient activation of high affinity nerve growth factor receptor, Trk, and morphological differentiation with fatal outcome in PC12 cells."Brain Res. 859. 169-172 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Fukumoto S, Mutoh T, Hasegawa T, Miyazaki H, Okada M, Goto G, Furukawa K, Urano T, Furukawa K.: "GD3 synthase gene expression in PC12 cells resulted in the continuous activation of TrkA and ERK1/2, leading to enhanced proliferation."J Biol Chem. 275. 5832-5838 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Mutoh T, Hamano T, Tokuda A, Kuriyama M.: "Unglycosylated Trk protein does not co-localize nor associate with gangliosides GM1 in stable clone overexpressing Trk of PC12 cells (PCtrk)."Glycoconj J. 17. 233-237 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Mutoh T, et al: "Glucosylceramide synthase inhibitor inhibits the action of nerve growth factor in PC12 cells"J Biol Chem. 273. 26001-26007 (1998)
• [Publications] Pitto M,Mutoh, et al: "Influence of endogenous GM1 ganglioside on Trk B activity in cultured cerebellar granule cells"FEBS Lett. 483. 93-96 (1998)
• [Publications] Mutoh T et al.: "Involvement of tyrosine phosphorylation in HMG-CoA reductase inhibitor-induced cell death in L6 myoblacts"FEBS Lett. 444. 85-89 (1999)
• [Publications] Mutoh T, et al: "Role of tyrosine phosphorylation of phospholipase C-γ1 in the signal transduction pathway of HMG-CoA reductase inhibitor-"FEBS Lett. 446. 91-94 (1999)
• [Publications] Fukumoto S,Mutoh T et al: "GD_3 Synthase gene expression in PC12 cells resulted in the continuous activation EKA and ERK1/2, leading to"J Biol Chem. 275. 5832-5838 (2000)
• [Publications] Mutoh T et al.: "Unglycosyluted Trk protein does not co-localize nor associate with ganglioside GM1 in stable clone"Glyconj J. 17. 233-237 (2000)
• [Publications] Mutoh T, Kumano T, Nakagawa H, Kuriyama M.: "Involvement of tyrosine phosphorylation and in HMG-CoA reductase inhibitor-induced cell death of L6 myoblasts."FEBS Lett. 444. 85-89 (1999)
• [Publications] Mutoh T, Kumano T, Nakagawa H, Kuriyama M.: "Role of tyrosine phosphorylation of phospholipase C-γ1 in the signal transduction pathway of HMG-CoA reductase inhibitor-induced cell death in L6 myoblasts."FEBS Lett. 446. 91-94 (1999)
• [Publications] Mitsuma N, Yamamoto M, Li M, Ito Y, Mitsuma T, Mutoh T, Takahashi M, Sobue G.: "Expression of GDNF receptor (Ret and GDNFR-α) mRNAs in the spinal cord of patients with amyotrophic lateral sclerosis."Brain Res. 820. 77-85 (1999)
• [Publications] Hamano T, Mutoh T, Hirayama M, Kuriyama M.: "MRI findings of benign monomeric amyotrophy of lower limb."J Neurol Sci. 165. 184-187 (1999)
• [Publications] Mutoh T, Sobue G, Hamano T, Kuriyama M, Hirayama M, Yamamoto M, Mitsuma T.: "Decreased tyrosine phosphorylation of TrkB receptor in the spinal cords from the patients with amyotrophic lateral sclerosis."Neurochem Res. (in press).
• [Publications] Hamano T, Mutoh T, Sugie H, Koga H, Kuriyama M.: "Phosphoglycerate kinase deficiency. An adult myopathic form with a novel mutation."Neurology. (in press).
• [Publications] Kumano T, Mutoh T, Nakagawa H, Kuriyama M.: "HMG-CoA reductase inhibitor induces a transient activation of high affinity nerve growth factor receptor, Trk, and morphological differentiation withfatal outcome in PC12 cells."Brain Res. (in press).
• [Publications] Fukumoto S, Mutoh T, Hasegawa T, Miyazaki H, Okada M, Goto G, Furukawa K, Furukawa K.: "GD3 synthase gene expression in PC12 cells resulted in the continuous activation of TrkA and ERK1/2, leading to enhanced proliferation."J Biol Chem. (in press).