PATHOGENESIS OF MOTOR NEURON DISEASE

运动神经元疾病的发病机制

• 批准号: 6112216
• 负责人: GAYLE E. WOLOSCHAK
• 金额: $ 22.98万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-06-01 至 2000-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6112216
• 关键词: animal genetic material tag; apoptosis; autosomal recessive trait; complementary DNA; degenerative motor system disease; enzyme activity; gene deletion mutation; genetic promoter element; genetically modified animals; immunogenetics; inborn immunodeficiency; laboratory mouse; motor neurons; neural degeneration; neurogenetics; oxidative stress; pathologic process; proliferating cell nuclear antigen; superoxide dismutase; thymus

Wasted mice bear an autosomal recessive gene (wst) that causes motor neuron degeneration, radiation sensitivity in T lymphocytes and immunodeficiency. Recent work in our laboratory has demonstrated a 3-bp (3G) deletional mutation in the promoter (5' region) of the PCNA gene of wasted mice that is not found in control littermates. We wish to test the hypothesis that abnormal regulation of PCNA expression caused by this 3G deletion is responsible for the wst mutation and the wasted phenotype of these mice. The specific aims of this proposal are: (1) To confirm the 3G deletion as the molecular basis for the motor neuron disease in wasted mice through the use of vector-based and transgenic approaches. (2) To identify the protein(s) [and their gene(s)] that bind specifically to the 11G element in the PCNA promoter, to map the protein binding regions, and to determine mechanisms underlying motor neuron-specific function. (3) To investigate the reasons for motor neuron death in the wasted mouse: a) studies of tissues from wasted mice in terms of pathology, and apoptosis, b) in vitro studies of neuronal cells from wasted mice (for PCNA expression, apoptosis, and oxidative damage), c) studies examining a possible common pathway of motor neuronal cell death in the mouse, and d) differential display of wasted and FALS transgenic mouse cDNA. (4) To sequence PCNA promoters in ALS, FALS, and other motor neuron diseases to identify possible mutations associated with the diseases and to analyze these disorders for sensitivity to oxidative stress and PCNA expression. These experiments will test aspects of the following model: absence of PCNA expression in thymus and motor neurons of wasted mice causes cellular apoptosis; this absence of expression of mediated by a positive transfactor which can bind to the wild-type but not the wasted mutant PCNA promoter; the bound protein induces late expression of PCNA in motor neurons and T lymphocytes; ALS and FALS patients have abnormalities in this pathway either via a common SOD/PCNA circuit or via differing SOD and PCNA pathways that result in apoptosis/cell death.

废弃小鼠携带常染色体隐性基因（wst），导致运动 神经元变性、T 淋巴细胞的辐射敏感性和 免疫缺陷。我们实验室最近的工作证明了 3-bp (3G)PCNA基因启动子(5'区)的缺失突变 在对照同窝小鼠中未发现浪费的小鼠。我们希望测试 假设该3G引起PCNA表达异常调节 缺失是造成 wst 突变和浪费表型的原因 这些老鼠。本提案的具体目的是： (1) 确认3G 缺失是废人运动神经元疾病的分子基础 通过使用基于载体和转基因方法的小鼠。 (2) 至 识别特异性结合的蛋白质[及其基因] PCNA 启动子中的 11G 元件，用于绘制蛋白质结合区域图，以及 确定运动神经元特异性功能的机制。 (3) 至 研究废弃小鼠运动神经元死亡的原因：a) 对废弃小鼠组织进行病理学和细胞凋亡研究， b) 废弃小鼠神经元细胞的体外研究（PCNA 表达、细胞凋亡和氧化损伤），c）研究检查 小鼠运动神经元细胞死亡的可能共同途径，以及 d) 废弃和 FALS 转基因小鼠 cDNA 的差异显示。 (4) 至 对 ALS、FALS 和其他运动神经元疾病中的 PCNA 启动子进行测序，以 识别与疾病相关的可能突变并进行分析 这些疾病对氧化应激和 PCNA 表达敏感。 这些实验将测试以下模型的各个方面： 废弃小鼠胸腺和运动神经元中 PCNA 的表达导致细胞 细胞凋亡；这种由阳性介导的表达缺失 可以结合野生型但不结合废弃突变体 PCNA 的转因子 发起人；结合蛋白诱导运动细胞中 PCNA 的晚期表达 神经元和T淋巴细胞； ALS 和 FALS 患者在以下方面存在异常： 该途径要么通过共同的 SOD/PCNA 电路，要么通过不同的 SOD 和 导致细胞凋亡/细胞死亡的 PCNA 途径。