Studies on functional xonation of adrenocortical cells

肾上腺皮质细胞功能性氧化的研究

• 批准号: 09044333
• 负责人: ISIMURA Yuzuru
• 金额: $ 1.15万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09044333/
• 关键词: adrenal cortex; mineralocorticoid; glucocorticoid; thymidine kinase; transgenic mice; SV40 large T antigen; Y-1 cells; AP-1 transcription factor; 副賢皮質

Mechanisms for the functional zonation of adrenocortical cells were studied using mice that were genetically manipulated. Findings obtained from research results are described below.1. Production of mice whose mineralocorticoid synthesizing cell-zone in the adrenal cortex are conditionally ablated. Thymidine kinase gene of herpes simplex virus was inserted into the locus at mineralocorticoid synthase gene of embryonic stem cells by transfection with a targeting vector. Recombinant embryonic stem cell clones were successfully obtained, and introduction of them into mouse blastocysts for chimeric animals are undergoing.2. Establishment of adrenocortical precursor cell-lines. Adrenocortical cells from transgenic mice which harbor an immortalizing gene encoding temperature sensitive SV40 large T antigen, were cultured and cloned with a conventional methods. The cell lines were characterized for expression of various steroidogenic genes by analyzing their mRNAs. The results indicated that a cell-line was undifferentiated at the permissive temperature and express glucocorticoid synthase gene at the nonpermissive temperature. Thus, and adrenocortical precursor cell-line which was able to differentiate upon a temperature shift was established.3. Molecular mechanisms by which adrenocorticotropic hormone (ACTH) stimulates glucocorticoid synthase gene expression. Using mouse adrenocortical Y-1 cells, ACTH was found to regulate transcription of the gene by causing compositional changes in AP-1 transcripotion factors in the cells via a cAMP-dependent pathway.4. Histochemical analyzes of mouse adrenal cortex. Immunohistochemical and in situ hybridization analyzes of mouse adrenal cortex showed the presence of an undifferentiated cell layr which express neither mineralonor glucocorticoid synthase.

机制的功能分区的肾上腺皮质细胞进行了研究，使用小鼠基因操纵。从研究结果中获得的发现如下：1。肾上腺皮质矿化皮质合成细胞带有条件消融小鼠的生产。用靶向载体转染单纯疱疹病毒胸苷激酶基因，将其插入胚胎干细胞矿糖皮质激素合成酶基因位点。成功地获得了重组胚胎干细胞克隆，并正在将其导入小鼠囊胚中用于嵌合动物。肾上腺皮质前体细胞的建立。采用常规方法培养和克隆了含有温度敏感型SV40大T抗原的转基因小鼠肾上腺皮质细胞。通过分析其mrna，鉴定了这些细胞系表达的各种类固醇基因。结果表明，细胞系在允许温度下未分化，在不允许温度下表达糖皮质激素合成酶基因。由此，建立了能随温度变化而分化的肾上腺皮质前体细胞系。促肾上腺皮质激素刺激糖皮质激素合成酶基因表达的分子机制。在小鼠肾上腺皮质Y-1细胞中，ACTH通过camp依赖途径引起细胞中AP-1转录因子的组成变化，从而调节该基因的转录。小鼠肾上腺皮质组织化学分析。小鼠肾上腺皮质的免疫组织化学和原位杂交分析显示存在未分化的细胞层，既不表达矿酸盐也不表达糖皮质激素合成酶。

项目成果

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Mitani,F. et al.: "Expression of Cytochromes P450aldo and P45011β in rat adrenal gland during late gestational and neonatal stages." Steroids. 62. 57-61 (1997)

Mitani, F. 等人：“妊娠晚期和新生儿阶段大鼠肾上腺中细胞色素 P450aldo 和 P45011β 的表达”。 62. 57-61 (1997)