Parkin and Its Regulation of Neuronal Apoptosis

Parkin及其对神经元凋亡的调节

• 批准号: 6791740
• 负责人: John Francois Staropoli
• 金额: $ 3.51万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6791740
• 关键词: Lentivirus; Parkinson' s disease; RNA interference; apoptosis; cyclins; dopamine; embryo /fetus; gene expression; gene mutation; genetic regulation; laboratory mouse; neural degeneration; neurons; neuropharmacology; neuroprotectants; neurotoxins; parkin gene /protein; predoctoral investigator; tissue /cell culture; transfection /expression vector; ubiquitin

DESCRIPTION (provided by applicant): Mutations in parkin underlie an autosomal recessive form of Parkinson's disease, the second most common neurodegenerative disease. To test a working model of parkin as a component of a multi-subunit, SCF-like ubiquitin ligase complex that protects dopamine neurons from apoptosis, other components of the complex, including sel-10 and cullin-1, will be dowregulated by RNA interference in murine primary neuronal cultures. Downregulation of these components will be evaluated for potentiation of dopamine neuron apoptosis and compared to the effects of downregulating parkin itself. To test the hypothesis that a candidate substrate of the parkin-associated complex, cyclin E, is a key mediator of the apoptotic cascade(s) against which wildtype parkin normally protects neurons, pharmacological inhibition of cyclin E-associated activity will be evaluated for rescue of dopamine neurons in the context of parkin, sel-10, or cullin-1 downregulation. Finally, lentivirus-mediated overexpression of parkin in the same primary culture system will be assessed for protection of dopamine neurons from neurotoxins as compared to overexpression of mutant forms of parkin, including clinically defined mutations and forms deleted in the ubiquitin homology and RING domains.

描述（由申请人提供）：帕金突变是帕金森病常染色体隐性遗传形式的基础，帕金森病是第二种最常见的神经退行性疾病。为了测试帕金蛋白作为多亚基SCF样泛素连接酶复合物的一个组成部分的工作模型，该复合物保护多巴胺神经元免于凋亡，该复合物的其他组成部分，包括sel-10和cullin-1，将通过RNA干扰在小鼠原代神经元培养物中下调。将评估这些组分的下调对多巴胺神经元凋亡的增强作用，并与下调parkin本身的作用进行比较。为了检验parkin相关复合物的候选底物细胞周期蛋白E是野生型parkin通常保护神经元的凋亡级联的关键介体的假设，将评价细胞周期蛋白E相关活性的药理学抑制以在parkin、sel-10或cullin-1下调的背景下拯救多巴胺神经元。最后，在相同的原代培养系统中，将评估慢病毒介导的parkin过表达与parkin突变形式的过表达相比对多巴胺神经元免受神经毒素的保护，包括临床定义的突变和在泛素同源性和RING结构域中缺失的形式。