Regulation of autophagy in dopaminergic cell death

多巴胺能细胞死亡中自噬的调节

• 批准号: 8066000
• 负责人: Charleen T Chu
• 金额: $ 28.39万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8066000
• 关键词: 1-Methyl-4-phenylpyridinium; 3-methyladenine; Abbreviations; Acute; Address; Antioxidants; Autophagocytosis; Biochemical; Brain Diseases; Cardiolipins; Cell Death; Cell Line; Cells; Chronic; Complex; Development; Dominant-Negative Mutation; Dopamine; Dopaminergic Cell; Dose; Equilibrium; Extracellular Signal Regulated Kinases; Genes; Green Fluorescent Proteins; Impairment; In Vitro; Injury; Lead; Lecithin; Lewy Body Disease; Life; Light; MAP Kinase Gene; MAP1 Microtubule-Associated Protein; MEKs; Mediating; Membrane; Metabolic; Metabolic Diseases; Midbrain structure; Mitochondria; Modeling; Molecular; Morphology; Mus; N-terminal; Nerve Degeneration; Neurites; Neurodegenerative Disorders; Neuronal Injury; Neurons; Neurotoxins; Nutrient; Organelles; Oxidants; Oxidation-Reduction; Oxidative Stress; Oxidopamine; Parkinson Disease; Pathologic; Phosphatidylethanolamine; Phosphatidylinositols; Phosphatidylserines; Phospholipid Signaling Pathway; Phospholipids; Phosphotransferases; Physiological; Play; Process; Protein Isoforms; Protein Kinase; Proteins; RNA Interference; Reactive Oxygen Species; Reagent; Regulation; Research Personnel; Role; Signal Transduction; Small Interfering RNA; Starvation; Stress; Substantia nigra structure; Superoxide Dismutase; System; Testing; Toxic effect; Toxin; Transgenic Organisms; Tyrosine 3-Monooxygenase; Vacuole; age related; cellular imaging; deprivation; design; dopaminergic neuron; extracellular; in vivo; in vivo Model; inhibitor/antagonist; injured; inorganic phosphate; insight; kinase inhibitor; mitochondrial autophagy; monodansylcadaverine; neurotoxic; novel; oxidation; phosphatidylethanolamine; programs; response; wortmannin

DESCRIPTION (provided by applicant): Dopaminergic (DA) neurons are sensitive to oxidative insults and degenerate in age-related neurodegenerative diseases. A morphologic form of regulated cell death characterized by prominent autophagic vacuoles (AVs) has been identified in neurons. Autophagy is normally a highly regulated process sequestering cytoplasmic components for lysosomal degradation. However, dysregulated or excessive autophagy can be harmful to cells, producing a condition that can be conceptualized as "autophagic stress." Although AVs are observed in degenerating DA neurons in Parkinson disease and its in vitro and in vivo models, the role of autophagy in DA neuronal injury remains to be elucidated. Our studies indicate that oxidative neurotoxins elicit increased mitochondrial autophagy in DA neurons. Moreover, the regulation of this injury-induced autophagy is different from that of nutrient-deprivation systems. This proposal investigates the hypotheses that: autophagy contributes to neurite retraction and cell death in injured DA neurons, and that reactive oxygen species and MARK signals regulate injury-induced autophagy. We will use the complex I inhibitor MPP+ to produce mitochondria-targeted injury, and the redox cycling 6- hydroxydopamine to model generalized oxidative stress, comparing acute and chronic treatments. A combination of molecular, biochemical, live cell imaging and transgenic approaches will be applied to DA cell lines, primary midbrain cultures and mice to determine the role of autophagy in DA neurite retraction and cell death, and to study MAPK and oxidative phospholipid signals involved in its regulation. Completion of these studies will yield important insights into mechanisms by which autophagic responses regulate DA neurite degeneration and cell death during oxidative neuronal injuries. Relevance: Mitchondrial impairment and autophagic stress are prominent features of Parkinson/Lewy body disease. In contrast to physiologic conditions, inducing autophagy in the presence of dysregulating pathologic forces may promote cell death. A better understanding of mechanisms that contribute to autophagic stress will help focus future research efforts to restore balance to the system. Thus, studying the role and regulation of autophagic responses in oxidatively-injured neurons may enhance development of novel therapies applicable to age-related neurodegenerative diseases and other brain disorders involving oxidative stress.

描述（由申请人提供）：多巴胺能（DA）神经元对氧化损伤敏感，并且在与年龄相关的神经退行性疾病中退化。在神经元中发现了一种以显着的自噬空泡（AV）为特征的受调节细胞死亡的形态形式。自噬通常是一个高度调控的过程，隔离细胞质成分以进行溶酶体降解。然而，失调或过度的自噬可能对细胞有害，产生一种可以概念化为“自噬应激”的状况。尽管在帕金森病及其体外和体内模型中的退化 DA 神经元中观察到 AV，但自噬在 DA 神经元损伤中的作用仍有待阐明。我们的研究表明，氧化神经毒素会引起 DA 神经元线粒体自噬增加。此外，这种损伤诱导的自噬的调节与营养剥夺系统的调节不同。该提案研究了以下假设：自噬导致受损 DA 神经元的轴突回缩和细胞死亡，活性氧和 MARK 信号调节损伤诱导的自噬。我们将使用复合物 I 抑制剂 MPP+ 产生线粒体靶向损伤，并使用氧化还原循环 6-羟基多巴胺来模拟全身氧化应激，比较急性和慢性治疗。将分子、生化、活细胞成像和转基因方法相结合，应用于 DA 细胞系、原代中脑培养物和小鼠，以确定自噬在 DA 神经突收缩和细胞死亡中的作用，并研究参与 DA 神经突收缩和细胞死亡的 MAPK 和氧化磷脂信号。其监管。这些研究的完成将对氧化神经元损伤期间自噬反应调节 DA 神经突变性和细胞死亡的机制产生重要的见解。相关性：线粒体损伤和自噬应激是帕金森/路易体病的突出特征。与生理条件相反，在病理力量失调的情况下诱导自噬可能会促进细胞死亡。更好地了解导致自噬应激的机制将有助于集中未来的研究工作以恢复系统的平衡。因此，研究氧化损伤神经元中自噬反应的作用和调节可能会促进适用于与年龄相关的神经退行性疾病和其他涉及氧化应激的脑部疾病的新疗法的开发。