MOLECULAR DYNAMICS OF RECEPTORS OF LIPID SOLUBLE SIGNAL MOLECULES

脂溶性信号分子受体的分子动力学

基本信息

  • 批准号:
    11470006
  • 负责人:
  • 金额:
    $ 9.41万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
  • 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

GFP-GR chimera construct was transfected into the cultured cells. By using luciferase assay GFP-GR chimera system was confirmed to be functional. DEX-inducible mouse mammary tumor virus promoter (MMTV)-Luc reporter was induced by DEX.Real-time imaging study has shown that in the absence of ligand GFP-GR was present in the cytoplasm of cells. Exposure of DEX to cultured cells induced the nuclear accumulation of GFP-GR and its translocation from cytoplasm-to-nucleus was time-dependent manner. Thirty min after DEX treatment GFP-GR was completely translocated into the nucleus. No green fluorescence was observed in the nucleolus. The same results were observed in the primary cultured hippocampal neurons and cortical glia. GFP-GR from the small cytoplasm of the perikarya to the nucleus was observed after 15 to 30 min after the DEX as well as corticosterone treatment, indicating that there was no conspicuous difference of trafficking manner of GR in the presence of the ligands among cell types. GFP-MR chimera construct was also transfected into COS-1 cells, hippocampal neurons, and cortical glia. As in the case of GFP-GR, GFP-MR chimera construct should be verified to be transcriptionally active before observing the real-time imaging. GFP-MR was observed in the cytoplasm of most of the COS-1 cells without the ligand. Aldosterone, an agonist for MR, induced the translocation of GFP-MR from the cytoplasm to the nucleus in a time-dependent manner. Within 30 min after the treatment green fluorescence was completely present in the nucleus and nucleolus was devoid of GFP-MR.Corticosterone treatment caused the similar accumulation of GFP-MR in the nucleus of the COS-1 cells where no fluorescence was observed at the nucleolus. Cells that were transfected by GFP-ERa construct and were not treated by estrogen showed the GFP-ERa in the nucleus. Nucleolus was devoid of GFP-ERa.
将 GFP-GR 嵌合体构建体转染至培养细胞中。通过使用荧光素酶测定,GFP-GR嵌合体系统被证实具有功能。 DEX诱导型小鼠乳腺肿瘤病毒启动子(MMTV)-Luc报告基因是由DEX诱导的。实时成像研究表明,在没有配体的情况下,细胞的细胞质中存在GFP-GR。将 DEX 暴露于培养细胞会诱导 GFP-GR 的核积累,并且其从细胞质到细胞核的易位呈时间依赖性方式。 DEX处理后三十分钟,GFP-GR完全转位到细胞核中。核仁中未观察到绿色荧光。在原代培养的海马神经元和皮质神经胶质细胞中观察到相同的结果。 DEX和皮质酮处理后15至30分钟后观察到GFP-GR从核周小细胞质到细胞核,表明配体存在下细胞类型间GR的运输方式没有明显差异。 GFP-MR嵌合体构建体也被转染到COS-1细胞、海马神经元和皮质神经胶质细胞中。与 GFP-GR 的情况一样,在观察实时成像之前,应验证 GFP-MR 嵌合体构建体是否具有转录活性。在大多数没有配体的 COS-1 细胞的细胞质中观察到 GFP-MR。醛固酮是 MR 的激动剂,以时间依赖性方式诱导 GFP-MR 从细胞质易位到细胞核。处理后 30 分钟内,绿色荧光完全出现在细胞核中,并且核仁中没有 GFP-MR。皮质酮处理导致 COS-1 细胞的细胞核中 GFP-MR 类似积累,但在核仁处没有观察到荧光。被GFP-ERa构建体转染且未经雌激素处理的细胞在细胞核中显示出GFP-ERa。核仁缺乏 GFP-ERa。

项目成果

期刊论文数量(66)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
ITO,T., MORITA,N., NISHI,M., KAWATA,M.: "IN VITRO AND IN VIVO IMMUNOCYTOCHEMISTRY FOR THE DISTRIBUTION OF MINERALOCORTICOID RECEPTOR WITH THE USE OF SPECIFIC ANTIBODY."NEUROSCI.RES.. 37. 173-182 (2000)
ITO,T.、MORITA,N.、NISHI,M.、Kawata,M.:“使用特定抗体进行盐皮质激素受体分布的体外和体内免疫细胞化学。”NEUROSCI.RES.. 37. 173-
  • DOI:
  • 发表时间:
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  • 影响因子:
    0
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  • 通讯作者:
Nishi M.et al: "Real-time imaging of glucocoriticoid receptor dynamics in living neurons and glial cells in comparison with non-neural cells"Eur.J.Neurosci.. 11. 1927-1936 (1999)
Nishi M.等人:“与非神经细胞相比,活体神经元和神经胶质细胞中糖皮质激素受体动力学的实时成像”Eur.J.Neurosci.. 11. 1927-1936 (1999)
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    0
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KAWATA Mitsuhiro其他文献

KAWATA Mitsuhiro的其他文献

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{{ truncateString('KAWATA Mitsuhiro', 18)}}的其他基金

Structural analysis of molecular and behavioral neuroendocrinology on the sexual difference and hormonal action in the nervous system
分子和行为神经内分泌学对神经系统性别差异和激素作用的结构分析
  • 批准号:
    24300128
  • 财政年份:
    2012
  • 资助金额:
    $ 9.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Steroid hormones and their receptor interaction by Raman spectrometer analysis
通过拉曼光谱仪分析类固醇激素及其受体相互作用
  • 批准号:
    22659040
  • 财政年份:
    2010
  • 资助金额:
    $ 9.41万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Interaction of genomic and non-genomic actions of hormones on the nervous system
激素对神经系统的基因组和非基因组作用的相互作用
  • 批准号:
    20240036
  • 财政年份:
    2008
  • 资助金额:
    $ 9.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Environmental Effects on the Dynamism of Receptors of Lipophilic Signal Molecules in the Brain and its Functional Significance
环境对脑内亲脂信号分子受体动态的影响及其功能意义
  • 批准号:
    16200026
  • 财政年份:
    2004
  • 资助金额:
    $ 9.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
DYNAMICS AND REGULATION OF NUCLEAR RECEPTORS OF LIPID SOLUBLE MOLECULE IDENTIFIED BY USING GFPS
GFPS鉴定脂溶性分子核受体的动力学和调控
  • 批准号:
    13470005
  • 财政年份:
    2001
  • 资助金额:
    $ 9.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
TRANSCRIPTION FACTOR INTERACTIONS REVEALED BY FRET
FRET 揭示的转录因子相互作用
  • 批准号:
    11358013
  • 财政年份:
    1999
  • 资助金额:
    $ 9.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
ROLE OF STEROID HORMONES ON THE CELLULAR COMMUNICATION : MOLECULAR, MORPHOLOGICAL, PHYSIOLOGICAL AND BEHAVIORAL MULTIDICIPLINARY APPROACH
类固醇激素对细胞通讯的作用:分子、形态学、生理学和行为多学科方法
  • 批准号:
    10044314
  • 财政年份:
    1998
  • 资助金额:
    $ 9.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
Effects of perinatal steroid hormonal changes on the cytoarchitecture of brain
围产期类固醇激素变化对大脑细胞结构的影响
  • 批准号:
    09480214
  • 财政年份:
    1997
  • 资助金额:
    $ 9.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Steroid receptors-multiforms and their significance on brain differentiation
多种形式的类固醇受体及其对大脑分化的意义
  • 批准号:
    07458257
  • 财政年份:
    1995
  • 资助金额:
    $ 9.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
AN APPLICATION OF IN SITU HYBRIDIZATION AND ANTISENSE OLIGONUCLEOTIDE METHOD TO VISUALIZE AND MANIPULATE GENE EXPRESSION OF STEROID HORMONE RECEPTOR PROTEINS IN THE NERVOUS SYSTEM
应用原位杂交和反义寡核苷酸方法可视化和操纵神经系统中类固醇激素受体蛋白的基因表达
  • 批准号:
    05454674
  • 财政年份:
    1993
  • 资助金额:
    $ 9.41万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)

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Role of glucocorticoid receptor-mediated mRNA decay in alcohol dependence
糖皮质激素受体介导的 mRNA 衰减在酒精依赖中的作用
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