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 SHIP2（含有肌醇 5-磷酸酶 SHIP 同工酶的 SH2 结构域）的 mRNA 在发育和成熟大鼠大脑中定位的研究（Kondo、Owada、SainoSaito）中，在胚胎阶段的心室生发区中检测到了它。随着出生后发育的进行，该表达在白质细胞（推测为少突胶质细胞）中明显表达，但在整个发育过程中神经元中没有显着表达。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了 PLD1mRNA 的表达，而 PLD2 mRNA 主要在假定的星形胶质细胞中表达，尽管 PLD2 的基因在出生后早期灰质中短暂表达。，假定的神经元。使用DBH启动子产生PI4激酶转基因小鼠，并使用嗜铬细胞的高尔基部分（SainoSaito，Ross）对膜运输的影响进行检查，尽管结果不是确定的。产生了 PAP2A 基因敲除小鼠，但敲除小鼠的表型并不明显（Kanoh，Kondo）。 FABP 基因敲除小鼠已成功完成，并对其表型进行了生化和形态学分析（Kondo、Owada、Spener）。成功克隆了PLAI，并在成年猴脑中检测了其mRNA的定位，结果在小脑颗粒和浦肯野细胞（Glomset、Goto、Kondo）中强烈表达。在大鼠脑和体外细胞（Irvine、Kondo）中检查了 PIPKI 和 PKPKII 的定位，发现 PIPKI 直接与 ADP-核糖基化因子 I 相互作用，并使用我们的 PI4K cDNA 和细胞负责高尔基体中 PIP 的合成生物学方法学。与Merida和Perrozzi通过电子邮件就DGK、PAP等PI相关分子的功能意义进行了深入讨论。