Gene Expression of Glial Cells

胶质细胞的基因表达

基本信息

批准号：
05808077
负责人：
KIRIHARA Tadashi
金额：
$ 1.22万
依托单位：
Soka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

2', 3'-Cycli-nucleotide 3'-phosphodiesterase (CNP) is expressed exclusively by oligodendrocytes in the central nervous system. CNP mRNA reaches a maximum level at the time of most active myelination. We have previously shown in mouse that two mRNA species are formed by alternative use of the two transcription start point (tsp) and by accompanying alternative splicing. The use of proximal tsp forms the larger mRNA (mRNA I) and the use of distal tsp forms the smaller mRNA (mRNA I) and the use of distal tsp froms the smaller mRNA (mRNA II) ; they encode CNP I and CNP II respectively.1. Two putative promoter regions of mouse CNP gene upstream of the two tsp were analyzed by transient transfection of C6 glioma cells with chloramphenicol acetyltramsferase gene constructs. We showed that these two regions have independent promoter activity.2. Structure and chromosomal localization of human CNP gene were determined. The presence of two mRNA species are indicated though northern analysis reveals an apparently single mRNA band. Spot blot hydbridization of flow-sorted human chromosomes showed that CNP gene is localized in chromosome 17.3. We examined whether the two mRNA species are detectable separately by RT-PCR during the embryonic and following neonatal stages of mouse brain development. Two primer combinations were used to amplify specific sequences of mRNA I and mRNA II respectivly. We showed that low levels of both mRNA species are present in the brain of mouse embryos at least after embryonic day 11. Both mRNA species reach a small peak around embryonic day 16. The results suggest that CNP plays an additional role in early stages of brain development when no oligodendrocytes are found. It was also noted that mRNA II is the more abundant message than mRNA I during these early stages of brain development.

2'，3'-Cycli-核苷酸3'-磷酸二酯酶（CNP）仅由中枢神经系统中的少突胶质细胞表达。在最活跃的髓鞘中，CNP mRNA达到最大水平。以前，我们在小鼠中表明，通过替代使用两个转录起点（TSP）以及伴随的替代剪接来形成两个mRNA物种。近端TSP的使用形成较大的mRNA（mRNA I），使用远端TSP形成较小的mRNA（mRNA I），并使用较小的mRNA的远端TSP（mRNA II）；它们分别编码CNP I和CNP II.1。通过用氯霉素乙酰氨基酯酶基因构建体的C6胶质瘤细胞瞬时转染C6神经胶质瘤细胞，分析了两个TSP上游小鼠CNP基因的两个推定启动子区域。我们表明，这两个区域具有独立的启动子活性2。确定人CNP基因的结构和染色体定位。尽管北部分析显示出明显的单个mRNA带，但表明了两个mRNA物种的存在。流动的人类染色体的斑点印迹脑化表明，CNP基因位于17.3染色体中。我们检查了在小鼠脑发育的胚胎和新生儿阶段，RT-PCR是否可以通过RT-PCR分别检测到这两个mRNA物种。两种底漆组合用于分别扩增mRNA I和mRNA II的特定序列。我们表明，至少在胚胎第11天之后，两个mRNA的大脑都存在于小鼠胚胎的大脑中。两个mRNA物种在胚胎第16天周围达到一个小峰。结果表明，CNP在大脑的早期阶段起着额外的作用。当未发现少突胶质细胞时发育。还指出，在大脑发育的这些早期阶段，mRNA II是比mRNA I更丰富的信息。