NOVEL OLIGODENDROCYTE PROTEIN--IMPLICATIONS FOR MS

新型少突胶质细胞蛋白——对多发性硬化症的影响

• 批准号: 2737839
• 负责人: BRIDGET SHAFIT-ZAGARDO
• 金额: $ 28.5万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2737839
• 关键词: animal tissue; confocal scanning microscopy; developmental neurobiology; electron microscopy; gene expression; immunocytochemistry; immunofluorescence technique; microtubule associated protein; monoclonal antibody; multiple sclerosis; myelination; neurogenesis; oligodendroglia; tissue /cell culture

Multiple Sclerosis (MS) is a paralyzing disease affecting young adults. While a number of experimental drugs are available to minimize the devastation of the disease's course the cause of MS is unknown. A central issue for the treatment of MS is whether oligodendrocytes or resident oligodendrocyte precursors have the potential to remyelinate axons following episodes of demyelination. We have identified a novel variant of human high molecular weight (HMW) microtubule-associated protein-2, designated MAP-2+13, and have generated monoclonal antibodies specific to this splice form. Immunocytochemistry with light and electron microcopy have demonstrated MAP-2+13 staining in human fetal oligodendrocytes during process extension and active myelination, and in numerous oligodendrocytes adjacent to a zone of demyelination in sections from MS lesions. MAP-2+13 is either minimally or not expressed in oligodendrocytes in the normal adult CNS. The hypothesis to best tested in this proposal is that MAP-2+13 is required for the elaboration of oligodendrocyte processes during myelination and that it can be used as a marker for myelinating and remyelinating oligodendrocytes. To determine whether MAP-2+13 expression parallels myelination within the developing CNS, a range of fetal ages will be examined by double-label immunofluorescence and Confocal microscopy, immunoblotting and electron microscopy. MAP-2+13 expression in MS lesions will be extensively examined to determine if the expression correlates with remyelination. Studies will correlate MAP-2+13 expression with the type of lesion and will be compared with age-matched, non neurologic sections. Rat progenitor cells and primary oligodendrocyte cultures will permit the study of MAP-2+13 in a well characterized in vitro system. Analysis will include how changes in MAP-2+13 expression correlate with process outgrowth and oligodendrocyte maturation. These studies permit the examination of a newly identified MAP-2 form in oligodendrocytes and the potential to gain insight into the extension of processes during myelination and disease. Attempts to identify developmentally regulated genes and to understand the regulation involved in the extension of myelinating processes is beneficial for both our understanding of normal CNS development and for treating demyelinating diseases such as Multiple Sclerosis.

多发性硬化症（MS）是一种影响年轻人的瘫痪疾病。虽然有一些实验性药物可以将疾病的破坏性降到最低，但多发性硬化症的病因尚不清楚。多发性硬化症治疗的一个中心问题是，在脱髓鞘发作后，少突胶质细胞或常驻少突胶质细胞前体是否具有轴突再髓鞘化的潜力。我们已经鉴定出一种新的人类高分子量（HMW）微管相关蛋白-2变异，命名为MAP-2+13，并产生了针对这种剪接形式的单克隆抗体。免疫细胞化学光镜和电镜显示，在过程延伸和髓鞘形成活动期间，人胎儿少突胶质细胞以及MS病变切片中脱髓鞘区附近的许多少突胶质细胞中都有MAP-2+13染色。MAP-2+13在正常成人中枢神经系统的少突胶质细胞中表达极少或不表达。本提案中最好验证的假设是，在髓鞘形成过程中，MAP-2+13对于少突胶质细胞的精细加工是必需的，并且它可以用作髓鞘形成和再髓鞘形成的少突胶质细胞的标记物。为了确定MAP-2+13的表达是否与发育中的中枢神经系统中的髓鞘形成平行，将通过双标记免疫荧光和共聚焦显微镜、免疫印迹和电子显微镜检查胎儿年龄范围。MAP-2+13在MS病变中的表达将被广泛检测，以确定其表达是否与髓鞘再生相关。研究将MAP-2+13的表达与病变类型相关联，并将与年龄匹配的非神经切片进行比较。大鼠祖细胞和原代少突胶质细胞培养将允许在体外系统中研究MAP-2+13。分析将包括MAP-2+13表达的变化如何与过程生长和少突胶质细胞成熟相关。这些研究允许在少突胶质细胞中检查新发现的MAP-2形式，并有可能深入了解髓鞘形成和疾病过程的延伸。试图确定发育调控基因并了解髓鞘形成过程延伸的调控，对于我们理解正常的中枢神经系统发育和治疗脱髓鞘疾病（如多发性硬化症）都是有益的。