Neuronal Expression of Hemoglobin in Multiple Sclerosis Cortex

多发性硬化症皮质中血红蛋白的神经元表达

• 批准号: 8446033
• 负责人: Ernest J Freeman
• 金额: $ 21.47万
• 依托单位: KENT STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-15 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8446033
• 关键词: Address; Aftercare; Apoptosis; Applications Grants; Area; Automobile Driving; Autopsy; Axon; Brain; Cell Culture Techniques; Cells; Confocal Microscopy; Cytosine; Disease; Electron Microscopy; Electron Transport; Employee Strikes; Epigenetic Process; Erythropoietin; Event; Free Radical Scavengers; Gene Proteins; Goals; Hemin; Hemoglobin; Hydrogen Peroxide; Image; In Situ Hybridization; Inflammation; Inflammation Mediators; Inflammatory; Investigation; Iron; Lead; Lesion; Life; Metabolic; Methylation; Mitochondria; Modification; Multiple Sclerosis; Mus; Myelin; Neurons; Oxidative Phosphorylation; Oxidative Stress; Oxygen; Patients; Pattern; Peroxonitrite; Population; Proteins; Proteomics; Public Health; Rattus; Regulation; Resolution; Respiration; Reverse Transcriptase Polymerase Chain Reaction; Role; Sampling; Site; Slice; Tissues; Up-Regulation; Western Blotting; bisulfite; cytokine; disability; dopaminergic neuron; environmental change; expression vector; gray matter; improved; inhibitor/antagonist; mind control; mitochondrial dysfunction; nervous system disorder; neuroblastoma cell; neuron loss; neuropathology; novel; overexpression; oxidative damage; promoter; protein expression; research study; response; surface enhanced laser desorption ionization; therapeutic target; treatment effect; white matter

DESCRIPTION (provided by applicant): Damage to and loss of neurons and axons is correlated with disability in multiple sclerosis (MS), thus neuroprotective therapies are urgently needed. In previous studies we have found altered expression of mitochondrial genes and proteins involved in respiration resulting in dysfunctional mitochondrial activity in postmortem MS cortex. Recently, we have shown that hemoglobin is upregulated in MS cortex and have localized hemoglobin expression to neurons in the MS brain. We have also found hemoglobin expression in primary cultures of neurons isolated from rat brain. This is a surprising and novel finding and provides support for our hypothesis that hemoglobin expression may be involved in neuronal respiration. The goal of this study is to understand the function of hemoglobin expression in neurons and its significance to neuropathology in MS. In the proposed study we will investigate the extent and localization of hemoglobin expression in different regions of the MS brain by in situ hybridization, RT-PCR, western blotting, confocal imaging, and electron microscopy of postmortem MS and control tissue. We will investigate the mechanisms which regulate hemoglobin expression in primary neuronal cultures and will assess the role of hemoglobin in neuronal respiration and oxidative phosphorylation by performing high resolution respirometry on neuroblastoma cells expressing hemoglobin subunits from a mammalian expression vector, and on brain slices from mice treated with erythropoietin which increases hemoglobin expression in the brain. We will also investigate the possibility that hemoglobin serves a neuroprotective role in the brain as a scavenger of free radicals. PUBLIC HEALTH RELEVANCE: Multiple sclerosis (MS) is a debilitating neurological disease which strikes patients in the prime of their life. We have previously shown that there is mitochondrial involvement in this disease. The proposed study is relevant to public health in that it will lead to a better understanding of mechanisms leading to altered mitochondrial activity in MS and will potentially identify new neuroprotective therapeutic targets.

描述（由申请人提供）：神经元和轴突的损伤和损失与多发性硬化症（MS）的残疾相关，因此迫切需要神经保护治疗。在以前的研究中，我们发现线粒体基因和蛋白质的表达改变，参与呼吸导致功能障碍的线粒体活性在死后MS皮质。最近，我们发现血红蛋白在MS皮质中上调，并在MS脑中的神经元中定位血红蛋白表达。我们还发现从大鼠脑分离的神经元的原代培养物中有血红蛋白表达。这是一个令人惊讶的和新颖的发现，并为我们的假设，血红蛋白的表达可能参与神经元呼吸提供了支持。本研究的目的是了解血红蛋白在神经元中表达的功能及其对多发性硬化症神经病理学的意义。在拟议的研究中，我们将通过原位杂交、RT-PCR、免疫印迹、共聚焦成像和死后多发性硬化症和对照组织的电子显微镜来研究多发性硬化症大脑不同区域血红蛋白表达的程度和定位。我们将研究调节原代神经元培养物中血红蛋白表达的机制，并通过对表达来自哺乳动物表达载体的血红蛋白亚基的神经母细胞瘤细胞和小鼠的脑切片进行高分辨率呼吸测量来评估血红蛋白在神经元呼吸和氧化磷酸化中的作用。用促红细胞生成素治疗，增加大脑中血红蛋白的表达。我们还将研究血红蛋白作为自由基清除剂在大脑中发挥神经保护作用的可能性。 公共卫生相关性：多发性硬化症（MS）是一种使人衰弱的神经系统疾病，在患者生命的黄金时期发作。我们以前已经表明，有线粒体参与这种疾病。这项拟议的研究与公共卫生有关，因为它将导致更好地了解导致MS中线粒体活性改变的机制，并可能确定新的神经保护治疗靶点。