MICROTUBULES AND MYELINATION

微管和髓鞘形成

• 批准号: 2270486
• 负责人: IAN DAVID DUNCAN
• 金额: $ 20.55万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-07-01 至 1998-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2270486
• 关键词: MICROTUBULES; MYELINATION

Myelination of the central nervous system (CNS) is a complex developmental process, requiring exquisitely timed interactions between axons and the myelin forming cells, the oligodendrocyte, and the appropriate temporal expression of an array of myelin genes. The initial elaboration of the myelin sheath in the developing CNS, and the long-term maintenance of compact myelin, each place different demands on the myelin forming cell. However, both processes are likely to be influenced by the structural and transport properties of the oligodendrocyte cytoskeleton. This proposal will study a new and unique rat myelin mutant, the taiep rat, which develops a chronic progressive neurological disease as a consequence of a CNS myelin disorder, associated with a microtubular accumulation in oligodendrocyte. The long-term goals are to characterize this mutant and its molecular defect and use it to define the role of microtubule and associated proteins in myelin formation and maintenance. To achieve these long-term goals, five specific aims have been planned: 1) The phenotype of the mutant will be characterize using morphological and morphometric approaches to study gla, cell kinetics, myelin, and the onset of the microtubule accumulation. 2) To determine whether environmental factors in the taiep CNS play any role in the development of the microtubule accumulation, cross transplantation experiments will be carried out. These will study the structure and function of taiep oligodendrocyte transplanted into the CNS of other myelin mutants, and of normal cells transplanted into the taiep CNS. 3) The polarity, stability and organizing centers of microtubule in the taiep rat oligodendrocyte will be compared to normal cells to determine whether an abnormal organization of tubules might interfere with transport events within the cell. 4) The role of microtubule regulatory proteins in the taiep mutation will be studied using antibodies to these proteins by Western blots and immunolabelling. Absence, or an over-or under- expression, or ectopic expression, of such a protein will be determined. Messenger RNAs for these proteins will also be examined by Northern blots and in situ hybridization. Finally, 5) To study the effects of accumulation of microtubule in taiep oligodendrocyte, cultures of these cells will be immunolabelled for myelin proteins known to co-localize with the cytoskeleton, and myelin protein mRNAs will be localized by in situ hybridization. Similar experiments will be performed on normal oligodendrocyte in which the cytoskeleton has been pharmacologically manipulated.

中枢神经系统(CNS)的髓鞘形成是一个复杂的过程 发展过程，需要精确计时的相互作用 轴突和髓鞘形成细胞、少突胶质细胞和 髓鞘基因阵列的适当时间表达。首字母 中枢神经系统发育过程中髓鞘的阐述及其长期意义 维持致密的髓鞘，每种对髓鞘的要求不同 正在形成细胞。然而，这两个过程都可能受到 少突胶质细胞细胞骨架的结构和运输特性。 该计划将研究一种新的、独特的大鼠髓鞘突变体taiep。 大鼠，它会发展为一种慢性进行性神经系统疾病 与微管相关的中枢神经系统髓鞘疾病的后果 在少突胶质细胞中堆积。长期目标是将 这个突变体及其分子缺陷，并用它来定义 髓鞘形成和维持中的微管和相关蛋白。 为实现这些长期目标，已规划了五个具体目标： 1)突变体的表型将用形态特征来描述。 和形态计量学方法来研究GLA、细胞动力学、髓鞘和 微管开始堆积。2)确定是否 大脑皮层中枢神经系统中的环境因素在发育中起着任何作用 对于微管的积累，交叉移植实验将 被执行。这些都将研究taiep的结构和功能。 将少突胶质细胞移植到其他髓鞘突变体的中枢神经系统中，以及 将正常细胞移植到大脑皮层中枢神经系统。3)极性， 大鼠睡眠时微管的稳定性和组织中心 少突胶质细胞将与正常细胞进行比较，以确定是否存在 小管的异常组织可能会干扰运输事件 在细胞内。4)微管调节蛋白在心肌梗死中的作用 将使用这些蛋白质的抗体通过以下方式研究taiep突变 免疫印迹和免疫标记。缺席，或过度或不足- 这种蛋白质的表达或异位表达将被确定。 这些蛋白质的信使RNA也将通过Northern blotts进行检测 和原位杂交。最后，5)研究 培养的少突胶质细胞中微管的堆积 细胞将被免疫标记已知的共同定位的髓鞘蛋白 随着细胞骨架的形成，髓鞘蛋白mRNAs将被定位于 原位杂交。类似的实验将在正常情况下进行 细胞骨架在药物作用下的少突胶质细胞 被操纵了。