FUNCTIONAL DOMAINS OF THE MYELIN BASIC PROTEINS

对髓鞘碱性蛋白（MBP）所起作用的完整理解 在组织髓鞘膜中的作用将阐明重要的机制 潜在的髓鞘修复。 髓鞘形成障碍突变体的可用性 老鼠，颤抖者，提供了一个独特的机会来研究 这一组蛋白质的髓鞘膜形成，因为 Shiverer缺乏功能性MBP基因，无法正常生成 致密的CNS髓鞘。 该提案旨在1）构建质粒 含有全长cDNA和截短的MBP， 转化为在啮齿动物细胞中有效表达的载体，2）评估 不同长度的MBP与shiverer单层髓鞘膜的结合 囊泡，和3）用载体转染颤抖少突胶质细胞 含有那些MBP构建体，这些MBP构建体已经显示出与膜结合， 囊泡 最终的目标是将这些转染的细胞移植到 同系颤抖小鼠获得髓鞘再生。 分子和细胞 生物技术将被用于以及免疫细胞化学在 光学和电子显微镜水平。 脱髓鞘在许多疾病引起的残疾中起着重要作用。 神经系统疾病，无论它们是否如MS中那样主要影响髓鞘，或 其次是中风和外伤。 彻底而批判性的检查 MBPs在髓鞘形成和稳定中的作用 膜将有望阐明机制，鼓励 膜的再生对正常的神经功能至关重要。

A complete understanding of the role the myelin basic proteins (MBPs) play in organizing the myelin membrane will elucidate important mechanisms underlying myelin repair. The availability of a dysmyelinating mutant mouse, the shiverer, affords a unique opportunity to study the contribution of this single set of proteins to myelin membrane formation, since the shiverer lacks a functional MBP gene and is unable to generate normally compacted CNS myelin. This proposal aims to 1) construct plasmids containing cDNAs of full length and truncated MBPs which will be inserted into vectors that are efficiently expressed in rodent cells, 2) evaluate binding of the varying-length MBPs to shiverer unilamellar myelin membrane vesicles, and 3) transfect shiverer oligodendrocytes with vectors containing those MBP constructs that have been shown to bind to membrane vesicles. The eventual goal is to transplant these transfected cells into syngenic shiverer mice to obtain remyelination. Molecular and cellular biological techniques will be used as well as immunocytochemistry at the light and electron microscopic level. Demyelination plays a significant role in the disability caused by many neurological diseases, whether they affect myelin primarily as in MS, or secondarily as in stroke and trauma. A thorough and critical examination of the role the MBPs play in the formation and stabilization of the myelin membrane will hopefully elucidate mechanisms that encourage the regeneration of a membrane so vital to normal neurologic function.