Genetic Dissection of Age-dependent Neuroprotection Mechanisms in Drosophila

果蝇年龄依赖性神经保护机制的遗传解析

基本信息

批准号：
8447484
负责人：
BARRY S GANETZKY
金额：
$ 35.59万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-15 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8447484
关键词：
Adolescence Advanced Glycosylation End Products Affect Autophagocytosis Biochemical Biological Biological Assay Brain Cells Cessation of life Chimeric Proteins Collection Coma Communication Defect Dihydroxyacetone Phosphate Dissection Drosophila genus Enzymes Event Exhibits Experimental Models Functional disorder Gene Proteins Generations Genes Genetic Genetic screening method Glyceraldehyde 3-Phosphate Glycolysis Goals Health Histological Techniques Human Link Longevity Lysosomes Maintenance Maleimides Maps Mediating Medical Membrane Fusion Molecular Molecular Analysis Molecular Genetics Motor Mutation Nerve Degeneration Neurodegenerative Disorders Neurons Oxidative Stress Paralysed Pathway interactions Phenotype Process Proteins Pyruvaldehyde Recycling Role Signal Transduction Surveys Synaptic Transmission Synaptic Vesicles System Testing Toxic effect Triose-Phosphate Isomerase age related base biological adaptation to stress crosslink dopaminergic neuron dosage fly in vivo mutant nervous system disorder neuromechanism neuron loss neuronal survival neuroprotection novel overexpression prevent progressive neurodegeneration protein complex relating to nervous system research study synuclein wasting

项目摘要

DESCRIPTION (provided by applicant): Our long-term aim is to elucidate molecular mechanisms of neural signaling in Drosophila. Here we focus on analysis of mutants with defects in maintenance of neuronal viability. We have discovered several mutations among our collection, including wstd and comt, that exhibit shortened lifespan and age-dependent, progressive neurodegeneration providing us with novel starting points to dissect neuroprotective mechanisms. Our goals are to determine the in vivo roles of the affected proteins using genetic, molecular, biochemical, and histological techniques to analyze how defects in these proteins result in the observed phenotypes. wstd encodes the glycolytic enzyme, triose phosphate isomerase (Tpi) responsible for the interconversion of DHAP (dihydroxyacetone phosphate) and GAP (glyceraldehyde 3-phosphate), only the latter of which is able to continue through glycolysis. Mutations of Tpi in humans result in Triosephosphate isomerase deficiency, characterized by early death and neurodegeneration but the underlying mechanism has remained unclear. We hypothesize that the enzymatic block in Tpi-deficient flies and humans leads to excess accumulation of methylglyoxal (MG), which reacts with target proteins to generate advanced glycation end products (AGEs) causing loss of protein activity, cross-linking, aggregation, and ultimately neuronal death. We propose genetic and biochemical experiments to test and refine this hypothesis. comt, which encodes NSF-1 (N-ethyl- maleimide sensitive fusion protein), exhibits a deficit in lysosomes and accumulation of ubiquitinated protein complexes in parallel with neurodegeneration. We hypothesize that comt is deficient in autophagy. Experiments are proposed to test this hypothesis and to dissect the step(s) in the process that are impaired. Additional mechanisms of neuroprotection will be investigated by phenotypic and molecular analysis of other mutants in our collection that exhibit neurodegeneration. Neuroprotective mechanisms are essential for proper neural communication and their disruption leads to some of the most devastating human neurological disorders. Detailed understanding of these mechanisms is thus of fundamental biological as well as medical importance. Drosophila has already proven to be a potent experimental system for elucidating these mechanisms. Moreover, both wstd and comt have direct links with human neurodegenerative disorders. Consequently, our proposed analyses should have broad biological and medical significance by contributing important new information that will advance our understanding of the underlying molecules and mechanisms that maintain neuronal viability and integrity.

描述（由申请人提供）：我们的长期目标是阐明果蝇神经信号传导的分子机制。在这里，我们重点分析在维持神经元活力方面存在缺陷的突变体。我们在我们的收藏中发现了几种突变，包括 wstd 和 comt，它们表现出寿命缩短和年龄依赖性、进行性神经变性，为我们剖析神经保护机制提供了新的起点。我们的目标是利用遗传、分子、生物化学和组织学技术来确定受影响蛋白质的体内作用，以分析这些蛋白质的缺陷如何导致观察到的表型。 wstd 编码糖酵解酶、磷酸丙糖异构酶 (Tpi)，负责 DHAP（磷酸二羟丙酮）和 GAP（3-磷酸甘油醛）的相互转化，只有后者能够继续进行糖酵解。人类 Tpi 突变会导致磷酸丙糖异构酶缺乏，其特征是过早死亡和神经变性，但其潜在机制仍不清楚。我们假设 Tpi 缺陷的果蝇和人类中的酶阻滞导致甲基乙二醛 (MG) 的过量积累，它与靶蛋白反应生成高级糖基化终产物 (AGE)，导致蛋白质活性丧失、交联、聚集和最终神经元死亡。我们提出遗传和生化实验来测试和完善这一假设。 comt 编码 NSF-1（N-乙基马来酰亚胺敏感融合蛋白），在神经变性的同时表现出溶酶体缺陷和泛素化蛋白复合物的积累。我们假设 comt 缺乏自噬。建议进行实验来检验这一假设并剖析过程中受损的步骤。我们将通过对我们收集的其他表现出神经变性的突变体进行表型和分子分析来研究其他神经保护机制。神经保护机制对于正常的神经通讯至关重要，其破坏会导致一些最具破坏性的人类神经系统疾病。因此，详细了解这些机制具有基本的生物学和医学重要性。果蝇已被证明是阐明这些机制的有效实验系统。此外，wstd 和 comt 都与人类神经退行性疾病有直接联系。因此，我们提出的分析应该具有广泛的生物学和医学意义，通过提供重要的新信息，这将增进我们对维持神经元活力和完整性的基本分子和机制的理解。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Death following traumatic brain injury in Drosophila is associated with intestinal barrier dysfunction.

DOI：
10.7554/elife.04790
发表时间：
2015-03-05
期刊：
eLife
影响因子：
7.7
作者：
Katzenberger RJ;Chtarbanova S;Rimkus SA;Fischer JA;Kaur G;Seppala JM;Swanson LC;Zajac JE;Ganetzky B;Wassarman DA
通讯作者：
Wassarman DA

NF-κB Immunity in the Brain Determines Fly Lifespan in Healthy Aging and Age-Related Neurodegeneration.