Analysis of the mechanism for neuronal degeneration and regeneration and responsible gene using the mutant mice disrupted of complex ganglioside

复合神经节苷脂破坏突变小鼠神经元变性再生机制及相关基因分析

• 批准号: 14570118
• 负责人: FURUKAWA Keiko
• 金额: $ 2.56万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570118/
• 关键词: glycosyltransferase; acidic glycosphingolipids; ganglioside; knock out; degeneration of nervous system; sensory dysfunction; DNA microarray; GM2; GD2 synthase gene; GD2; 神経; 複合型糖脂質; 糖類合成酵素遺伝子; DNAチップ; cDNAsubtraction法

We have cloned a number of glycosyltransferase cDNA, products of which are acidic glycosphingolipids (ganglioside), expressed mainly in the nerve system. We have also generated knock out (KO) mice to analyze the biological function of gangliosides. As a result, we found that complex gangliosides play an important role for survival and maintenance of the nervous system. In this study, we analyzed the genes, expression levels of which are different between the wild type and GM2/GD2 synthase gene KO mice using a comprehensive appriache DNA microarray.We analyzed the chronological alteration of phenotypes in GM2/GD2 synthase gene KO mice and found the degeneration of nervous tissues in the dorsal horn of the spinal cord. Neurological examination revealed progressive sensory dysfunction with aging. We compared the mRNA expression in the spinal cord between the wild type and 20 weeks-old male KO mice by DNA microarray. mRNA expression levels of TGTP, IFI47,IFIT1,IRF7 and IRF1 were higher in KO mice than in the wild type mice. However, the difference in the expression levels was not significant. In the future, we will investigate the relation between these interferon-induced genes and the degeneration of the nervous system, and analyze the expression levels of these genes in older mice after onset of nervous system degeneration. Furthermore, we will prepare the RNA from degenerated sites in the spinal cord then analyze the gene expression profiles in the spinal cord of the wild type and KO mice.

我们已经克隆了一些糖基转移酶cDNA，其产物是酸性鞘糖脂（神经节苷脂），主要在神经系统中表达。我们还产生了基因敲除（KO）小鼠来分析神经节苷脂的生物学功能。因此，我们发现复合神经节苷脂对神经系统的生存和维持起着重要作用。本研究利用基因芯片技术，分析了野生型和GM 2/GD 2合酶基因敲除小鼠中表达水平不同的基因，分析了GM 2/GD 2合酶基因敲除小鼠表型的时间变化，发现了脊髓背角神经组织的变性。神经系统检查发现随着年龄的增长，感觉功能障碍逐渐加重。我们通过DNA微阵列比较了野生型和20周龄雄性KO小鼠脊髓中mRNA的表达。TGTP、IFI 47、IFIT 1、IRF 7和IRF 1的mRNA表达水平在KO小鼠中高于野生型小鼠。然而，表达水平的差异不显著。今后，我们将研究这些干扰素诱导的基因与神经系统退化之间的关系，并分析这些基因在老年小鼠神经系统退化后的表达水平。此外，我们将从脊髓中的变性位点制备RNA，然后分析野生型和KO小鼠脊髓中的基因表达谱。

项目成果

Fukawa, K.: "Isolation and functional analysis of the melanoma specific promoter region of human GD3 synthase gene"Biochim.Biophys.Acta.. 1627. 71-78 (2003)

Fukawa，K.：“人GD3合酶基因的黑色素瘤特异性启动子区域的分离和功能分析”Biochim.Biophys.Acta..1627.71-78(2003)

Nakamura, Y.: "Identification of a Drosophila gene encoding xylosylprotein β4-galactosyltransferase that is essential for the synthesis of glycosaminoglycans and for morphogenesis"J Biol Chem.. 277. 46280-46288 (2002)

Nakamura, Y.：“编码木糖基蛋白 β4-半乳糖基转移酶的果蝇基因的鉴定，该酶对于糖胺聚糖的合成和形态发生至关重要”J Biol Chem.. 277. 46280-46288 (2002)

Iwamura, K.: "The blood group P1 synthase gene is identical to the Gb3/CD77 synthase gene : A solution of the P1/P2/p puzzle."J.Biol.Chem.. 278. 44429-44438 (2003)

Iwamura, K.：“血型 P1 合酶基因与 Gb3/CD77 合酶基因相同：P1/P2/p 难题的解决方案。”J.Biol.Chem.. 278. 44429-44438 (2003)

Bullens, R.W.M.: "Complex gangliosides at the neuromuscular junction are essential receptors for autoantibodies and botulinum neurotoxin but redundant for normal synaptic function."J.Neurosci. 22. 6876-6884 (2002)

Bullens, R.W.M.：“神经肌肉接头处的复合神经节苷脂是自身抗体和肉毒杆菌神经毒素的重要受体，但对于正常突触功能来说是多余的。”J.Neurosci。

Nakamura, Y.: "Identification of a Drosophila gene encoding xylosylprotein beta4-galactosyl-transferase that is essential for the synthesis of glycosamino-glycans and for morphogenesis"J. Biol. Chem.. 277. 46280-46288 (2002)

Nakamura，Y.：“编码木糖基蛋白β4-半乳糖基转移酶的果蝇基因的鉴定，该酶对于糖胺聚糖的合成和形态发生至关重要”J.