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.

在一项研究（Kondo，Owada，Sainosaito）关于Ship2的MRNA定位的研究中，SH2-域含有肌醇5-磷酸酶船在发育和成熟大鼠的大脑中，在胚胎阶段的心室生发区中检测到它。随着产后发育的进行，这种表达在白质细胞中很明显，该细胞的少突胶质细胞的假定，在整个发育过程中没有明显的神经元表达。 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.使用DBH启动器生成了用于PI4-激酶的转基因小鼠，并使用铬脂细胞（Sainosaito，Ross）进行膜转运效果的检查，尽管结果尚未确认。生成了PAP2A的基因敲除小鼠，但敲除小鼠的表型并不明显（Kanoh，Kondo）。成功完成了用于Fabps的基因敲除小鼠，并且在生化和形态上已经对其表型进行了分析（Kondo，Owada，Spener）。 PLAI的cDNA克隆成功，并在猴子的成年大脑中检查了其mRNA的定位，从而导致小脑颗粒和珀kinje细胞的强烈表达（Glomset，Goto，kondo）。在大鼠脑和体外细胞（Irvine，kondo）和Pipki中检查了PIPKI和PKPKII的定位，并发现使用我们的PI4K cDNA和细胞生物学方法论直接与ADP-核糖基化因子I I直接相互作用，并负责Golgi c室中的PIP合成。通过电子邮件就DGK，PAP以及其他与Merida和Perrozzi的其他PI相关分子的功能意义进行了亲密讨论。