Regulation of Neuroprotective Lipid Sgnals by Spatially Distinct NMDA Receptors

空间上不同的 NMDA 受体对神经保护性脂质信号的调节

• 批准号: 8231538
• 负责人: David Thoms Stark
• 金额: $ 3.4万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8231538
• 关键词: ALOX15 gene; Action Potentials; Affect; Agonist; Alzheimer' s Disease; Anti-Inflammatory Agents; Anti-inflammatory; Apoptosis; Apoptotic; Arachidonate 15-Lipoxygenase; Attenuated; Bathing; Biochemistry; Biological Availability; Biomedical Research; Brain; Brain-Derived Neurotrophic Factor; Calcium; Cell Culture Techniques; Cell Death; Cell Survival; Cells; Cellular biology; Cessation of life; Cyclic AMP-Responsive DNA-Binding Protein; Dependence; Development; Disease; Docosahexaenoic Acids; Enzymes; Epilepsy; Equilibrium; Exhibits; Fluo-3; Functional disorder; Gene Expression; Genes; Health; Healthcare; High Pressure Liquid Chromatography; Hippocampus (Brain); Human; Image; In Vitro; Injury; Ischemia; Link; Lipids; Lipoxygenase 1; Measures; Mediating; Mediator of activation protein; Mitochondria; Modeling; Molecular; Molecular Biology; Mus; N-Methyl-D-Aspartate Receptors; Necrosis; Neurodegenerative Disorders; Neurons; Outcome; Pathology; Pathway interactions; Patients; Physiological; Process; Protocols documentation; Receptor Signaling; Regulation; Reperfusion Therapy; Research; Retinal; Role; Signal Transduction; Societies; Stroke; Synapses; Techniques; Testing; Touch sensation; Toxic effect; Wild Type Mouse; base; brain cell; care burden; excitotoxicity; improved; in vivo; lipid mediator; mitochondrial dysfunction; neuronal survival; neuroprotectin D1; neuroprotection; novel; programs; research study; response; tandem mass spectrometry

DESCRIPTION (provided by applicant): The proposed research is part of a long-term effort to understand how lipid signaling mechanisms determine the fate of brain cells affected by diseases which exhibit excitotoxic cell death. Stimulation of synaptic NMDA receptors promotes neuronal survival, while stimulation of extrasynaptic NMDA receptors leads to dominant, pro-death signaling. However, the molecular pathways that determine these two outcomes are not fully understood. Two largely non-overlapping gene expression programs mediating cell survival and cell death, respectively, are induced by signaling from these spatially distinct NMDA receptor subpopulations. Neuroprotection D1 (NPD1) is a stereospecific lipid mediator derived from docosahexaenoic acid (DMA) that activates a pro-survival gene expression program in human brain and retinal cells. Does NPD1 mediate pro- survival signals from synaptic NMDA receptors? Control of NPD1 bioavailability by synaptic and extrasynaptic NMDA receptors will be investigated using primary hippocampal cell cultures. Experiments will focus on NPD1 pool size regulation by brain-derived neurotrophic factor (BDNF), which is a mediator of pro- survival signaling from synaptic NMDA receptors and a known agonist of NPD1 synthesis. Experiments will employ techniques from cell and molecular biology and biochemistry, HPLC-ESI tandem mass spectrometry-based lipidomic analysis, and the use of 15-lipoxygenase-1 (15-LOX-1) deficient mice, which are incapable of NPD1 synthesis. The results of the proposed studies will demonstrate that pro-survival signaling from the synaptic NMDA receptor is mediated through enhancement of the NPD1 pool size via increased expression of BDNF. Additionally, 15-LOX-1 deficient neuronal cultures will be shown to be significantly more vulnerable to excitotoxic injury than wild type cultures, and NPD1 will be shown to attenuate excitotoxic injury in vitro and in vivo. PUBLIC HEALTH RELEVANCE: Stroke, epilepsy, and neurodegenerative diseases such as Alzheimer's Disease, which share calcium toxicity as a common pathological feature, represent tremendous health care burdens on society and touch countless lives globally. Understanding the balance of molecular level processes leading to cell death or survival is critical to improving patient outcomes in these diseases, and neuroprotective signaling of DMA-derived lipid messengers is a new and important horizon on the biomedical research landscape.

描述（由申请人提供）： 这项研究是长期努力的一部分，旨在了解脂质信号传导机制如何决定受兴奋性细胞死亡疾病影响的脑细胞的命运。突触NMDA受体的刺激促进神经元存活，而突触外NMDA受体的刺激导致显性的促死亡信号传导。然而，决定这两种结果的分子途径尚未完全了解。来自这些空间上不同的NMDA受体亚群的信号传导分别诱导了两种基本上不重叠的基因表达程序，介导细胞存活和细胞死亡。神经保护D1（NPD 1）是一种立体特异性脂质介质，来源于二十二碳六烯酸（DMA），激活人脑和视网膜细胞中的促生存基因表达程序。NPD 1介导来自突触NMDA受体的促存活信号吗？将使用原代海马细胞培养物研究突触和突触外NMDA受体对NPD 1生物利用度的控制。实验将集中于脑源性神经营养因子（BDNF）对NPD 1池大小的调节，BDNF是来自突触NMDA受体的促存活信号传导的介体，并且是NPD 1合成的已知激动剂。实验将采用细胞和分子生物学和生物化学技术，基于HPLC-ESI串联质谱的脂质组学分析，以及使用15-脂氧合酶-1（15-LOX-1）缺陷小鼠，这些小鼠不能合成NPD 1。所提出的研究的结果将证明，促生存信号从突触NMDA受体介导的NPD 1池的大小通过增加BDNF的表达增强。此外，15-LOX-1缺陷型神经元培养物将显示出比野生型培养物显著更易受兴奋性毒性损伤，并且NPD 1将显示出在体外和体内减弱兴奋性毒性损伤。公共卫生关系：中风、癫痫和阿尔茨海默病等神经退行性疾病都具有钙毒性这一共同病理特征，给社会带来了巨大的医疗保健负担，并触及全球无数生命。了解导致细胞死亡或存活的分子水平过程的平衡对于改善这些疾病的患者结局至关重要，而DMA衍生的脂质信使的神经保护性信号传导是生物医学研究领域的一个新的重要领域。