Analysis of Neurosteroid Synthesis in Grial Cells with a combined Imaging System

利用组合成像系统分析胶质细胞中的神经类固醇合成

• 批准号: 08680713
• 负责人: OHTA Yoshihiro
• 金额: $ 1.47万
• 依托单位: Tokyo University of Agriculture and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08680713/
• 关键词: glial cells; neurosteroid; cholesterol; adrenocortical cells; steroid hormone; ACTH; low density lipoprotein; imaging; 副賢皮質細胞; カルシウム信号; シトクロムP450_<SCC>; 共焦点顕微鏡

To elucidate the mechanism of neurosteroid synthesis, we have visualized the process of intracellular signaling in glial cells and adrenocortical cells.Visualization was performed with he combination of confocal microscopy and real-time video enhanced microscopy. Since adrenal cortex is a major organ which produces steroid hormone, adrenocortical cells were examined as a model system of neurosteroid synthesis in glial cells.After stimulation of adrenocortical cells with ACTH,a series of calcium spikes and steroidogenesis were observed in the cells. There was a relationship between calcium signaling and steroid horm one synthesis, because inhibition of calcium signaling stopped steroidogenesis. The uptake of a low density lipoprotein (LDL) which contains a number of cholesterol molecules, precursors of steroid hormone, was observed. The trafficking of internalized LDL and perinuclear distribution of the free cholesterol were also observed.ATP is reported to be a neurotrasmitter. The stimulation of glial cells by ATP induced a transient calcium spikes in the cells. Glial cells also showed a intensive uptake of LDL and a directed trafficking of endosomes as adrenocortical cells showed. On the other hand, free cholesterol molecules in a glial cell were mainly located in a plasma membrance.

为了阐明神经甾体合成的机制，我们用共聚焦显微镜和实时视频增强显微镜观察了神经胶质细胞和肾上腺皮质细胞内信号传导的过程。由于肾上腺皮质是产生类固醇激素的主要器官，因此，以肾上腺皮质细胞作为神经胶质细胞合成神经类固醇的模型系统，用ACTH刺激肾上腺皮质细胞后，观察到细胞内一系列钙峰和类固醇合成。钙信号和类固醇激素合成之间存在关系，因为钙信号的抑制会停止类固醇激素的合成。观察到低密度脂蛋白（LDL）的吸收，其中含有许多胆固醇分子（类固醇激素的前体）。还观察到内化的低密度脂蛋白的运输和游离胆固醇的核周分布。据报道，ATP是一种神经递质。ATP刺激胶质细胞可引起细胞内瞬时钙峰。胶质细胞也表现出对LDL的强烈摄取和内体的定向运输，如肾上腺皮质细胞所示。另一方面，胶质细胞中的游离胆固醇分子主要位于质膜中。

项目成果

Kimoto,T.,et al.: "Digital Fluorescence Imaging of Elementary Steps of Neurosteroid Synthesis in Rat Brain Glial Cells" J.Pharmaceutical and Biochemical Analysis. (in press). (1997)

Kimoto,T.,et al.：“大鼠脑胶质细胞神经类固醇合成基本步骤的数字荧光成像”J.Pharmaceutical and Biochemical Analysis。

Kimoto,T., et al: "Advenocortricotropin Induces Calcium Oscillations in Aclrenal Fasciculata cells : Single cell Imaging" Biochem. Biophys. Res. Commun.221. 25-30 (1996)

Kimoto,T. 等人：“促肾上腺皮质激素在肾上腺束状细胞中诱导钙振荡：单细胞成像”Biochem。

川戸 佳,他: "脳グリア細胞と副腎皮質細胞のカルシウム信号のイメージング" 蛋白質核酸酵素. 42. 1227-1232 (1997)

Yoshi Kawato 等人：“脑胶质细胞和肾上腺皮质细胞中的钙信号成像”蛋白质核酸酶。42. 1227-1232 (1997)

Kimoto, T., et al: "Digital Fluorescence Imaging of Elementary steps of Neurosteroid Synthesis in Rat Brain Glial cells" J.Pharm.Biomed Anal. 15. 1231-1240 (1997)

Kimoto, T. 等人：“大鼠脑胶质细胞神经类固醇合成基本步骤的数字荧光成像”J.Pharm.Biomed Anal。

Kimoto, T., et al: "Digital Fluorescence Imaging of Elementary steps of Neurosteroid Synthesis in Rat Bratn Glial Cells" J.Pharm.Biomed.Anal.15. 1231-1240 (1997)

Kimoto, T. 等人：“大鼠 Bratn 胶质细胞中神经类固醇合成基本步骤的数字荧光成像”J.Pharm.Biomed.Anal.15。