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Molecular and Celluler Biological Analysis of the functional Significance of Phosphoinositide Metabolism

磷酸肌醇代谢功能意义的分子和细胞生物学分析

基本信息

批准号：
11694235
负责人：
KONDO Hisatake
金额：
$ 8.51万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

In a study (Kondo, Owada, SainoSaito) on the localization of mRNA for SHIP2, SH2-domain containing inositol 5-phosphatase SHIP isozyme in the brain of developing and mature rats, it was detected in the ventricular germinal zone at embryonic stages. As the postnatal development proceeded, the expression was evident in cells of the white matters, presumptive of oligodendrocytes, without significant expression in neurons throughout the development. In another study (Kondo, Owada, SainoSaito) on the localization of PLD mRNAs, the expression of PLD1mRNA was detected in presumptive oligodendrocytes, while PLD2 mRNA was expressed mainly in presumptive astrocytes, although the gene for PLD2 was expressed transiently in early postnatal gray matters, presumptive neurons. The transgenic mice for PI4kinases was generated using a DBH-promoter and the examination on the effect in membrane transport was performed using the Golgi fraction of chromaffin cells (SainoSaito, Ross), although the results are not confirmative. The gene knockout mice for PAP2A was generated but the phenotype of knockout mice was not evident (Kanoh, Kondo). The gene knockout mice for FABPs was successfully done and their phenotypes have been analyzed biochemically and morphologically (Kondo, Owada, Spener). The cDNA cloning of PLAI was succeeded and the localization of its mRNA was examined in the adult brain of monkey, resulting in the intense expression in the cerebellar granule and Purkinje cells (Glomset, Goto, Kondo). The localization of PIPKI and PKPKII was examined in the rat brain and in vitro cells (Irvine, Kondo) and PIPKI was found to directly interact with ADP-ribosylation factor I and be responsible for PIP synthesis in the Golgi compartment using our PI4K cDNA and cell biological methodologies. Intimate discussion was made through E-mail about the functional significance of DGK, PAP and some other PI-related molecules with Merida and Perrozzi.

在一项关于SHIP 2（含SH 2结构域的肌醇5-磷酸酶SHIP同工酶）mRNA在发育和成熟大鼠脑中的定位研究（Kondo，Owada，SainoSaito）中，在胚胎阶段的脑室生发区检测到SHIP 2。随着出生后发育的进行，在白色物质的细胞中表达明显，推测为少突胶质细胞，在整个发育过程中在神经元中没有显著表达。在另一项关于PLD mRNA定位的研究（Kondo，Owada，SainoSaito）中，在假定的少突胶质细胞中检测到PLD 1 mRNA的表达，而PLD 2 mRNA主要在假定的星形胶质细胞中表达，尽管PLD 2基因在出生后早期灰质（假定的神经元）中短暂表达。使用DBH-启动子产生PI 4激酶的转基因小鼠，并使用嗜铬细胞的高尔基体部分（SainoSaito，Ross）进行膜转运效果的检查，尽管结果不确定。产生了PAP 2A的基因敲除小鼠，但敲除小鼠的表型不明显（Kanoh，Kondo）。成功地进行了FABPs基因敲除小鼠，并对它们的表型进行了生化和形态学分析（Kondo，Owada，Sunday）。成功克隆了PLAI的cDNA，并对其mRNA在成年猴脑中的定位进行了研究，结果表明PLAI在小脑颗粒细胞和浦肯野细胞（Glomset，后藤，Kondo）中有强表达。在大鼠脑和体外细胞（Irvine，Kondo）中检查PIPKI和PKPKII的定位，并且发现PIPKI与ADP-核糖基化因子I直接相互作用，并且使用我们的PI 4K cDNA和细胞生物学方法负责高尔基体隔室中的PIP合成。通过E-mail与Merida和Perrozzi就DGK、PAP等PI相关分子的功能意义进行了深入讨论。