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

在Kondo，Owada，SainoSaito等人的研究中，SHIP2，SH2结构域含肌醇5-磷酸酶SHIP同工酶在发育和成熟大鼠脑中的mRNA定位研究中，在胚胎期的脑室生发区检测到。随着出生后发育的进行，在白质细胞中有明显的表达，推测为少突胶质细胞，在整个发育过程中神经元中没有明显的表达。在另一项关于PLD mRNAs定位的研究(Kondo，Owada，SainoSaito)中，PLD1mRNA在推测的少突胶质细胞中表达，而PLD2mRNA主要在推测的星形胶质细胞中表达，尽管PLD2基因在出生后早期的灰质(推测的神经元)中短暂表达。使用DBH启动子建立了PI4激酶转基因小鼠，并使用嗜铬细胞的高尔基组分(SainoSaito，Ross)对膜转运的影响进行了检测，尽管结果并不确定。产生了PAP2A基因敲除小鼠，但敲除小鼠的表型不明显(Kanoh，Kondo)。成功构建了FABPs基因敲除小鼠，并对其表型进行了生化和形态分析(近藤、小和田、Spener)。成功地克隆了Plai基因，并对其在成年猴脑中的表达进行了检测，结果在小脑颗粒和浦肯野细胞(Glomset、Goto、Kondo)中有较强的表达。PIPKI和PKII在大鼠脑和体外细胞(Irvine，Kondo)中的定位被发现，PIPKI直接与ADP-核糖化因子I相互作用，并负责高尔基体内PIP的合成。通过电子邮件与Merida和Perrozzi就DGK、PAP等PI相关分子的功能意义进行了深入的讨论。

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.