Involvement in neurodegeneration and cell death of synuclein family protein

1)我们培育了α- synnuclein转基因（α-syn Tg）小鼠，但野生小鼠与α-syn Tg小鼠在表型上无显著差异。2)采用α-syn Tg小鼠，进行DNA阵列分析。DNA阵列分析结果表明α-突触核蛋白可能影响细胞死亡。将人α-突触核蛋白和突变体cdna引入IMR-32细胞。这些细胞用维生素K3、顺铂、紫杉醇和视黄酸等各种刺激处理。α-synuclein对野生和转化的IMR-32细胞无明显影响。3)为了明确哺乳动物突触核蛋白家族蛋白的功能，对其在两栖和爬行动物等低等动物中的功能进行了研究。对蛇脑和蛙脑裂解液进行2d凝胶电泳，经胰蛋白酶凝胶酶切后用LC-MS/MS测定部分氨基酸序列。在蛇和蛙体内分别鉴定出2个和1个突触核蛋白与牛β-突触核蛋白泛抗体具有交叉反应性。

1) We have produced α-synuclein transgenic (α-syn Tg) mice but there was no significant difference in phenotype between wild and α-syn Tg mice.2) Using α-syn Tg mouse, analysis by DNA array was carried out. The result of DNA array analyzed showed that α-synuclein might affect cell death. Human α-synuclein and mutant cDNAs were introduced in IMR-32 cells. Those cells were treated with various stimuli such as vitamin K3, cisplatin, taxol and retinoic acid. However, α-synuclein did not significantly affect cell death induced by those stimuli in wild and transformed IMR-32 cells.3) To clarify the function of mammal synuclein family proteins, investigation of their in lower animals such as amphibian and reptile was performed. Brain lysates prepared from snake and frog were subjected to 2D-gel electrophoresis and then partial amino acid sequences were determined by LC-MS/MS after in gel digestion with trypsin. Two and one synuclein proteins shown the crossreactivity with pan-antibody produced against bovine β-synuclein were identified in snake and frog, respectively.