Molecular mechanism for the maintenance and repair of nervous sptem with acidic

酸性维持和修复神经系统的分子机制

• 批准号: 12670111
• 负责人: FURUKAWA Keiko
• 金额: $ 2.05万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670111/
• 关键词: ganglioside; knock-out; targeting; neuro-degeneration; regeneration; subtraction; neuron; glia; 遺伝子発現; GM2; GD3; 酸性スフィンゴ糖脂質; 糖鎖合成酵素遺伝子; ノックアウトマウス; 行動異常; グリア細胞; 神経細胞

1. For the mutant mice of GM2/GD2 Synthase gene lacking complex gangliosides (1) pathological analysis on neuro-degeneration with aging, and (2) analysis on molecular mechanisms for the degeneration were performed. First of all, for the peripheral nerve degeneration marked degeneraion of sciatic nerves, apoptosis of neurons in the dorsal root ganglia, death of neurons in the dorsal horn of the spinal cord, and enlargement of glia cells were found. In the analysis with electron microscopy, extension and enlargement of astrocytes and engulfment of blood vessels with ie projection of astrocytes were observed. Furthermore, morpholoicafchanged in the synaptic vesicles and synapse formation (spine type or dendrite type) were detected. These abnorrX changes became definite in the mutant mice after 35 weeks old (male) or 45 weeks oldI fSmaleV and neurological abnormal findings such as gait disturbance were simultaneously found.nd 4S RNA was extracted from the mutaut and wild type mice at approximately 25 weeks old, when theneurological abnormalities and pathological changes were not yet obvious, to analyze the changes in the expression levels of genes in the mutant mice. Preparing cDNAs, subtraction and differential hybridization were performed, resulting in the detection of about 30 colnes shich showed marked down-regulation in the mutant mice. Among them, genes associated with mitochondrial membrane enzymes, protease genes, and transciption factor genes were included. Implication of those gene products in the degeneration is now under investigation.2. For the mutant mice of GD3 synthase gene lacking b-series gangliosides, (1) process of neuro- degeneration and (2) alteration in the regeneration potential were analyzed. La the mutant mice degeneration of the peripheral nerves was not definite, but marked reduction in the regeneration of lesioned nerves.

1.对GM2/GD2合酶基因突变小鼠缺乏复合神经节苷脂进行了(1)神经退行性变的病理分析和(2)退行性变的分子机制分析。首先，周围神经变性明显，坐骨神经变性，背根神经节神经元凋亡，脊髓背角神经元死亡，胶质细胞增大。电子显微镜下可见星形胶质细胞伸展增大，血管被星形胶质细胞突起吞噬。此外，还观察到突触小泡和突触形成(棘型或树突型)的形态变化。突变小鼠在35周龄(雄性)或45周龄时出现明显的异常X变化，同时发现步态障碍等神经系统异常表现。在神经学异常和病理变化尚不明显的情况下，从约25周龄的突变和野生型小鼠中提取ND 4S RNA，分析突变小鼠基因表达水平的变化。制备cDNA，进行消减和差异杂交，检测到约30个Colnes shich在突变小鼠中明显下调。其中包括与线粒体膜酶相关的基因、蛋白水解酶基因和转录因子基因。这些基因产物在退化中的作用目前正在调查中。对缺失b-系列神经节苷脂的GD3合成酶基因突变小鼠，分析了(1)神经变性过程和(2)再生潜能的改变。突变小鼠周围神经变性程度不明确，但受损神经再生明显减少。

项目成果

Furukawa, K: "Novel functions of complex carbohydrates elucidated by the mutant mice of glycosyltransferase genes"Bichim. Biophys. Acta.. 1525. 1-12 (2001)

Furukawa, K：“糖基转移酶基因突变小鼠阐明的复合碳水化合物的新功能”Bichim。

Ito,M et al: "Specificity of carbohydrate structures of gangliosides in the activity to regenerate the rat axotomized hypoglossal nerve."Glycobiology. 11. 125-130 (2001)

Ito,M 等人：“神经节苷脂碳水化合物结构在大鼠轴突舌下神经再生活性中的特异性。”糖生物学。

Furukawa, K el al: "Expression of the Gb3/CD77 synthase gene in megakaryoblastic leukemia cells : implication in the sensitivity to verotoxins"J. Biol. Chem. (in press).

Furukawa, K 等人：“巨核细胞白血病细胞中 Gb3/CD77 合酶基因的表达：对维罗毒素敏感性的影响”J.

Iwamoto, T.: "Lactosylceramide is essential for the osteoclastogenesis mediated by the macrophage-colony stimulating factor and receptor activator of NF-KB ligand"J. Biol. Chem.. 276. 46031-46038 (2001)

Iwamoto, T.：“乳糖神经酰胺对于巨噬细胞集落刺激因子和 NF-KB 配体受体激活剂介导的破骨细胞生成至关重要”。

Itoh, M et al: "Enhancement of rat hypoglossal nerve regeneration by chitin sheet plus-ganfiJiosides."Br. J. Plast. Surg.. 53. 607-611 (2000)

Itoh, M 等人：“甲壳素片加甘菲苷增强大鼠舌下神经再生。”Br。