Functional analysis of mouse 3-phosphoglycerate dehydrogenase (Phgdh) gene by using gene targeting

利用基因打靶技术对小鼠 3-磷酸甘油酸脱氢酶 (Phgdh) 基因进行功能分析

• 批准号: 14580756
• 负责人: FURUYA Shigeki
• 金额: $ 2.3万
• 依托单位: The Institute of Physical and Chemical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580756/
• 关键词: serine; glia; amino acid; neural stem cells; knockout mouse; metabolic defect; brain development; embryonic lethality; 神経栄養因子; 細胞増殖; 脳皮質形成; 脳; グリア細胞; アストロサイト

D-3-Phosphoglycerate dehydrogenase (Phgdh ; EC 1.1.1.95) is the first committed enzyme of L-serine biosynthesis in the phosphorylated pathway. To determine the physiological importance of Phgdh-dependent L-serine biosynthesis in vivo, we generated Phgdh-deficient mice using targeted gene disruption in embryonic stein cells. The absence of Phgdh led to a drastic reduction of L-serine metabolites such as phosphatidyl-L-serine (PS) and sphingolipids. Phgdh null embryos have small bodies with abnormalities in selected tissues and died after days post-coitum 13.5. Striking abnormalities were evident in the central nervous system in which the Phgdh null mutation culminated in hypoplasia of the telencephalon, diencephalon, and mesencephalon ; in particular, the olfactory bulbs, ganglionic eminence, and, cerebellum appeared as indistinct structures. These observations demonstrate that the Phgdh-dependent phosphorylated pathway is essential for normal embryonic development, especially for brain morphogenesis.

D-3-磷酸甘油酸脱氢酶（Phgdh；EC 1.1.1.95）是磷酸化途径中 L-丝氨酸生物合成的第一个关键酶。为了确定体内 Phgdh 依赖性 L-丝氨酸生物合成的生理重要性，我们利用胚胎 stein 细胞中的靶向基因破坏产生了 Phgdh 缺陷小鼠。 Phgdh 的缺失导致 L-丝氨酸代谢物如磷脂酰-L-丝氨酸 (PS) 和鞘脂的急剧减少。 Phgdh 无效胚胎的身体较小，在选定的组织中存在异常，并且在性交后几天后死亡 13.5。中枢神经系统出现了明显的异常，其中 Phgdh 无效突变最终导致端脑、间脑和中脑发育不全；特别是，嗅球、神经节隆起和小脑呈现出模糊的结构。这些观察结果表明，Phgdh 依赖性磷酸化途径对于正常胚胎发育，特别是大脑形态发生至关重要。

项目成果

Yamashita, N., Sakai, K., Furuya, S., Watanabe, M.: "Selective expression of L-serine synthetic enzyme 3PGDH in Schwann cells perineuronal glia and endoneurial fibroblasts in rat sciatic nerves and its upregulation after crush injury."Arch.Histol.Cytol..

Yamashita, N.、Sakai, K.、Furuya, S.、Watanabe, M.：“L-丝氨酸合成酶 3PGDH 在大鼠坐骨神经的雪旺细胞神经周围胶质细胞和神经内成纤维细胞中的选择性表达及其在挤压损伤后的上调。”

Yoshida, K., Furuya, S., Osuka, S., Mitoma, J., Shinoda, Y., Watanabe, M., Azuma, N., et al.: "Targeted disruption of the mouse 3-phosphoglycerate dehydrogenase gene causes severe neuro-developmental defects and results in embryonic lethality."J.Biol.Chem

Yoshida, K.、Furuya, S.、Osuka, S.、Mitoma, J.、Shinoda, Y.、Watanabe, M.、Azuma, N. 等人：“靶向破坏小鼠 3-磷酸甘油酸脱氢酶基因

Shimizu, M., Furuya, S., Shinoda, Y.et al.: "Functional analysis of mouse3-phosphoglycerate dehydrogenase(Phgdh) gene promoter in developing brain."J.Neurosci.Res. In press. (2004)

Shimizu, M.、Furuya, S.、Shinoda, Y.等人：“小鼠 3-磷酸甘油酸脱氢酶 (Phgdh) 基因启动子在大脑发育中的功能分析。”J.Neurosci.Res。

Chen, Peng, C., Luff, J., Spring, K., Watters, D., Bottle, S., Furuya, S., Lavin, M.F.: "Oxidative stress is responsible for deficient survival and dendritogenesis in Purkinje neurons from Atm mutant mice."J.Neurosci.. 23. 11453-11460 (2003)

Chen, Peng, C.、Luff, J.、Spring, K.、Watters, D.、Bottle, S.、Furuya, S.、Lavin, M.F.：“氧化应激是导致浦肯野神经元存活和树突生成不足的原因”

Chen, N., Furuya, S., Shinoda, Y.et al.: "Extracellular carbohydrate-signal triggering PKA-dependent neuronal actin-reorganization."Neuroscience. 122. 985-995 (2003)

Chen, N.、Furuya, S.、Shinoda, Y. 等人：“细胞外碳水化合物信号触发 PKA 依赖性神经元肌动蛋白重组。”神经科学。