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: 8539856
• 负责人: Ernest J Freeman
• 金额: $ 17.76万
• 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 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8539856
• 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.

描述（申请人提供）：多发性硬化症（MS）中神经元和轴突的损伤和丢失与残疾相关，因此迫切需要神经保护治疗。在之前的研究中，我们发现线粒体基因和呼吸相关蛋白的表达改变导致死后MS皮层线粒体活性失调。最近，我们发现多发性硬化症皮层中的血红蛋白上调，并且在多发性硬化症脑中的神经元中也有局部的血红蛋白表达。我们还发现从大鼠脑分离的神经元原代培养中有血红蛋白表达。这是一个令人惊讶的新发现，并为我们的假设提供了支持，即血红蛋白表达可能参与神经元呼吸。本研究的目的是了解多发性硬化症神经元中血红蛋白表达的功能及其在神经病理学中的意义。本研究将通过MS死后和对照组织的原位杂交、RT-PCR、western blotting、共聚焦成像和电镜研究多发性硬化症脑组织中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中血红蛋白表达的机制，并将通过对来自哺乳动物表达载体的表达血红蛋白亚基的神经母细胞瘤细胞和经促红细胞生成素处理的小鼠脑切片进行高分辨率呼吸测量，评估血红蛋白在神经元呼吸和氧化磷酸化中的作用。促红细胞生成素可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道夫在大脑中起到神经保护作用的可能性。