MOLECULAR STUDIES OF A NOVEL OLIGODENDROCYTE

描述：少突胶质细胞特异性蛋白（OSP）是最近分离的一种 中枢神经系统髓鞘特异性蛋白，其功能和调节表达过程中， 申请人拟解决的CNS发育和神经胶质肿瘤发生。 OSP在大脑中的发育表达将被表征， 原位杂交，以确定少突胶质细胞 蛋白质可能起作用的分化。 的启动子区 OSP基因将被分离和表征，以确定因素 调节这种基因的细胞特异性表达。 该分析将 在细胞培养和转基因小鼠中进行。 几行 证据支持OSP在调节少突胶质细胞增殖中的作用。 因此，OSP基因剂量对少突胶质细胞生长和增殖的影响可能与OSP基因剂量有关。 将检查髓鞘形成。 少突胶质细胞前体将是 用含有OSP cDNA的逆转录病毒表达载体转染， OSP反义cDNA，以确定过度和不足的影响 OSP的表达对细胞生长和分化的影响。 转基因小鼠将 用髓鞘碱性蛋白启动子驱动的OSP转基因产生 以促进OSP过表达，并将建立OSP“敲除”小鼠， 确定OSP低表达的影响。 这些突变小鼠将 通过免疫组织化学、原位杂交和电子显微镜分析， 显微镜，以揭示形态，结构和生化效应 OSP基因剂量的改变。 申请人及其同事亦会 通过利用酵母菌鉴定OSP相互作用的蛋白质 双混合动力系统 最后，他们将检查OSP的表达， 在一些少突胶质细胞瘤中OSP基因的拷贝数，试图 研究其在肿瘤发生中的潜在作用。

DESCRIPTION: Oligodendrocyte-specific protein (OSP) is a recently isolated CNS myelin-specific protein, whose function and regulated expression during CNS development and glial tumorigenesis the applicant proposes to address. OSP's developmental expression in the brain will be characterized using in situ hybridization, to determine at which stage in oligodendrocyte differentiation the protein is likely to act. The promoter region of the OSP gene will be isolated and characterized in order to identify factors that regulate the cell-specific expression of this gene. This analysis will be performed in cell culture and in transgenic mice. Several lines of evidence support a role of OSP in regulating oligodendrocyte proliferation. Therefore, the effect of OSP gene dosage on oligodendroglial growth and myelination will be examined. Oligodendrocyte precursors will be transfected with a retroviral expression vector containing the OSP cDNA and OSP anti-sense cDNAs in order to determine the effect of over and under expression of OSP on cell growth and differentiation. Transgenic mice will be created with an OSP transgene driven by the myelin basic protein promoter to promote OSP overexpression, and OSP "knock-out" mice will be created to determine the effect of OSP underexpression. These mutant mice will be analyzed by immunohistochemistry, in situ hybridization, and electron microscopy, to reveal the morphological, structural, and biochemical effects of alterred OSP gene dosage. The applicant and his colleagues will also identify the proteins with which OSP interacts by exploiting the yeast two-hybrid system. Finally, they will examine expression of OSP and the copy number of the OSP gene in a number oligodendrogliomas, in an attempt to investigate its potential role in tumorigenesis.