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: 6343898
• 负责人: BRIDGET SHAFIT-ZAGARDO
• 金额: $ 30.24万
• 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/6343898
• 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。虽然有许多实验药物可以将疾病过程的破坏降到最低，但MS的原因尚不清楚。治疗多发性硬化症的一个中心问题是，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有可能使轴突重新髓鞘。我们已经鉴定了一种新的人类高分子量(HMW)微管相关蛋白-2的变体，命名为MAP-2+13，并产生了针对这种剪接形式的单抗。免疫细胞化学光镜和电子显微镜显示MAP-2+13在突起延伸和活跃髓鞘形成过程中的人胎儿少突胶质细胞中表达，并在MS病变切片中邻近脱髓鞘区域的大量少突胶质细胞中表达。MAP-2+13在正常成人中枢神经系统少突胶质细胞中低表达或不表达。在这项建议中最好检验的假设是，MAP-2+13是髓鞘形成过程中少突胶质细胞加工所必需的，它可以作为髓鞘形成和重新髓鞘形成的少突胶质细胞的标志。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，将采用双标记免疫荧光和共聚焦显微镜、免疫印迹和电子显微镜对一系列胎儿年龄进行检测。MAP-2+13在MS皮损中的表达将被广泛检测，以确定其表达是否与重新髓鞘形成有关。研究将MAP-2+13的表达与病变类型相关联，并将其与年龄匹配的非神经科切片进行比较。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表达的变化如何与突起生长和少突胶质细胞成熟相关。这些研究允许检查新发现的少突胶质细胞中的MAP-2形式，并有可能深入了解髓鞘形成和疾病过程中突起的延伸。试图识别发育调控基因和了解髓鞘形成过程的延长所涉及的调控，对于我们理解中枢神经系统的正常发育和治疗多发性硬化症等脱髓鞘疾病都是有益的。