Mouse Models of Parkin Biology and Pathobiology

帕金生物学和病理学小鼠模型

• 批准号: 6842148
• 负责人: Ted M. Dawson
• 金额: $ 32.19万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6842148
• 关键词: Parkinson' s disease; affinity chromatography; alpha synuclein; biological signal transduction; disease /disorder model; environmental toxicology; gene mutation; gene targeting; genetically modified animals; high performance liquid chromatography; laboratory mouse; methylphenyltetrahydropyridine; model design /development; nerve /myelin protein; neural degeneration; parkin gene /protein; pathologic process; posttranslational modifications; protein protein interaction; protein structure function; proteomics; transfection

Mutations in the parkin gene are the main genetic cause of autosomal recessive Parkinson's disease (PD) and mutations in parkin also play a major role in familial PD. Preliminary studies indicate a potential pivotal role for parkin in the ubiquitin proteasomal pathway (UPP) by functioning as an ubiquitin E3 ligase. Most disease causing mutations of parkin are thought to be loss of function mutations that ultimately lead to the absence of ubiquitination and the subsequent failure of UPP-mediated degradation of parkin substrates. Thus, the abnormal accumulation of parkin substrates is thought to play a role in the demise of substantia nigra dopaminergic neurons in patients with parkin mutations. A number of putative parkin substrates have been identified, but their importance in the pathogenesis of PD due to parkin mutations is not known. We propose to characterize parkin knockout mice to formally test the hypothesis that the absence of parkin function is the cause of PD due to parkin mutations. Furthermore, biochemical and proteomic characterization of the parkin knockout mice may shed light on the substrates that are important in the pathogenesis of PD due to parkin mutations. Accordingly experiments are proposed to further characterize the role of parldn and it's substrates in the pathogenesis of PD. In Specific Aim #1 we will characterize parkin knockout mice. In Specific Aim #2 we will evaluate the sensitivity of parkin knockouts to environmental toxins. In Specific Aim #3 we will evaluate the interaction of parkin with the alpha-synuclein interacting protein, synphilin-1 and determine whether parkin mediates K48 or K63 ubiquitin linkages. In Specific Aim #4 we will determine whether parkin interacts with alpha-synuclein by evaluating of the effect of crossing parkin knockout mice with A53T mutant alpha-synuclein transgenic mice and further evaluate the interaction of parkin with the alpha-synuclein interacting protein, synphilin-1. In Specific Aim #5 we will identify and characterize parkin interacting proteins and identify compensatory changes in parkin knockout mice. Development and characterization of parkin knockout, understanding the relationship of parkin, alpha-synuclein and synphilin- 1 in the pathogenesis of PD may provide insight into the molecular mechanisms by which these gene products induce neuronal damage and may provide novel therapeutics and targets to prevent the toxic effects of these familial associated genes in the degenerative process of PD.

parkin基因突变是常染色体隐性遗传性帕金森病（PD）的主要遗传原因，parkin基因突变在家族性PD中也起着重要作用。初步研究表明，帕金在泛素蛋白酶体途径（UPP）中作为泛素E3连接酶发挥作用的潜在关键作用。大多数引起帕金蛋白突变的疾病被认为是功能丧失突变，其最终导致泛素化的缺失， 随后UPP介导的帕金底物降解失败。因此，帕金底物的异常积累被认为在帕金突变患者黑质多巴胺能神经元的死亡中起作用。一些假定的帕金底物已被确定，但其在帕金森病的发病机制，由于帕金突变的重要性是未知的。我们建议描述parkin基因敲除小鼠的特征，以正式验证这一假设 parkin功能缺失是parkin突变导致PD的原因。此外，parkin基因敲除小鼠的生物化学和蛋白质组学特征可能揭示了由于parkin突变导致PD发病的重要底物。因此，实验提出进一步表征parldn及其底物在PD发病机制中的作用。在具体目标#1中，我们将表征parkin敲除小鼠。在具体目标#2中，我们将评估parkin基因敲除对环境毒素的敏感性。在具体目标#3中，我们将评估parkin与α-突触核蛋白相互作用蛋白synphilin-1的相互作用，并确定parkin是否介导K48或K63泛素连接。在具体目标#4中，我们将通过评价parkin敲除小鼠与A53 T突变体α-突触核蛋白转基因小鼠的杂交效果来确定parkin是否与α-突触核蛋白相互作用，并进一步评价parkin与α-突触核蛋白相互作用蛋白synphilin-1的相互作用。 在具体目标#5中，我们将鉴定和表征parkin相互作用蛋白，并鉴定parkin敲除小鼠的代偿性变化。parkin基因敲除的发展和特征，了解parkin、α-突触核蛋白和synphilin- 1在PD发病机制中的关系，可能有助于深入了解这些基因敲除的分子机制。 基因产物诱导神经元损伤，并可能提供新的治疗和目标，以防止这些家族相关基因在PD的退行性过程中的毒性作用。