Intracellular signal transduction mechanims of neuronal survival and death

神经元生存和死亡的细胞内信号转导机制

• 批准号: 11670616
• 负责人: MUTOH Tatsuro
• 金额: $ 2.3万
• 依托单位: Fukui Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670616/
• 关键词: 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

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.

我们前期的研究表明，神经节苷脂GM 1（gangliosideGM 1）是膜筏的主要成分，它调节高亲和力神经生长因子受体Trk，而Trk是神经元分化和存活的关键因子。我们构建了缺失可能的N-糖基化位点的突变型人trk cDNA，并转染缺失Trk表达的PCnnr细胞。我们已经能够获得稳定的过表达突变体trk cDNA的转染子，该突变体克隆在酪氨酸自磷酸化反应、神经突起生长和防止血清剥夺诱导的细胞凋亡方面对NGF没有任何反应。这些突变体Trk蛋白不位于脂筏中，而是分布在蔗糖密度梯度超离心的较高密度级分中。为了验证上述数据，我们还检查了胆固醇拮抗剂如β-甲基环糊精和菲律宾肽对NGF诱导的Trk自磷酸化反应的影响，因为这些试剂已被报道破坏筏结构。我们没有观察到这些药物预处理的细胞对NGF的任何反应。这些数据有力地表明，GM 1和膜筏结构的Trk受体蛋白的正常功能的重要因素。

项目成果

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)

Hamano T、Mutoh T、Sugie H、Koga H、Kuriyama M.：“磷酸甘油酸激酶缺乏症。具有新突变的成人肌病形式。”神经学。

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

Fukumoto S、Mutoh T、Hasekawa T、Miyazaki H、Okada M、Goto G、Furukawa K、Urano T、Furukawa K.：“PC12 细胞中 GD3 合酶基因的表达导致 TrkA 和 ERK1/2 的持续激活，从而导致

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)

Mutoh T 等人：“酪氨酸磷酸化参与 HMG-CoA 还原酶抑制剂诱导的 L6 成肌细胞细胞死亡”FEBS Lett。

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)

Mutoh T、Kumano T、Nakakawa H、Kuriyama M.：“磷脂酶 C-γ1 酪氨酸磷酸化在 HMG-CoA 还原酶抑制剂诱导的 L6 成肌细胞细胞死亡的信号转导途径中的作用。”FEBS Lett。 94 (1999)

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)

Fukumoto S、Mutoh T 等人：“PC12 细胞中的 GD_3 合酶基因表达导致 EKA 和 ERK1/2 持续激活，从而导致”J Biol Chem.