| Project/Area Number |
05808077
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurochemistry/Neuropharmacology
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| Research Institution | Soka University |
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Principal Investigator |
KIRIHARA Tadashi Institute of Life Science, Soka University Professor, 生命科学研究所, 教授 (50018800)
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| Co-Investigator(Kenkyū-buntansha) |
SAIGO Kaoru Faculty of Science, University of Tokyo Professor, 理学部, 教授 (50136454)
HATTORI Kazue Institute of Life Science, Soka University Assistant Professor, 生命科学研究所, 講師 (70228485)
泉 龍太郎 創価大学, 生命科学研究所, 助手 (30247291)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1994: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1993: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Gene expression / Glial cell / Oligodendrocyte / CNP / Mouse / Human / Embryo / 胎生期 / オリゴデンドロサイト / 選択スプライシング / RT-PCR / 遺伝子 / 相同遺伝子 / プロモター |
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| Research Abstract |
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.
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