Establishment of cell models on transfection of functional protein and the study on brain signal transduction

功能蛋白转染细胞模型的建立及脑信号转导研究

基本信息

  • 批准号:
    12557011
  • 负责人:
  • 金额:
    $ 8.58万
  • 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:
    2000
  • 资助国家:
    日本
  • 起止时间:
    2000 至 2001
  • 项目状态:
    已结题

项目摘要

The present study intends to establish the model of the cell line in which functionally important proteins such as receptors, enzymes, ion channels are expressed. Stimulation of the established cells with neurotransmitters hormones, growth factors etc. activates the expressed proteins in the cells. These models serve as the established systems to analyze effects of drugs and to create new drugs.The study focuses the analysis on the intracellular calcium (Ca^<2+>) and on the functional significance of CaM kinases I, II, III, IV and kinase. The cytosolic and nuclear isoforms of CaM kinase II were expressed in NG108-15 cells in which the expression of brain-derived neurotrophic factor (BDNF) was examined. The expression of BDNF was not increased by the transfection of the cytosolic isoforms such as CaM kinase II δ1, and δ2 but was increased by CaM kinase II δ3 and αB. The quantitative method of RT-PCR for the amount of the mRNA indicated the increase in Exon IV-BDNF mRNA but not Exon III-BDNF mRNA. This suggest that the nuclear isoforms only increase the mRNA and proteins of BDNF.Dopamine D2 receptors (D2R) consist of the long chain receptor (D2RL) and the short chain receptor (D2RS). Both cDNAs were transfected into NG108-15 cells and the stable cells with the expression of each receptor were obtained. The analysis by confocal laser microscopy revealed different localization of each receptor in the cells. Stimulation of D2R increased the concentration of the intracellular Ca^<2+>. The cDNA of CaM kinase II δ3 with nuclear localization signal was transiently transfected in the stable cells of D2RL. Stimulation of D2R of the transfected cells with the agonist resulted in activation of the nuclear isoform of CaM kinase II and the increase in the expression of BDNF.
本研究旨在建立具有重要功能的蛋白质如受体、酶、离子通道等在细胞系中表达的模型。用神经递质、激素、生长因子等刺激已建立的细胞,激活细胞中表达的蛋白质。这些模型作为既定的系统来分析药物的效果和创造新的药物。本研究重点分析细胞内钙(Ca^<2+>)和CaM激酶I、II、III、IV和激酶的功能意义。在检测脑源性神经营养因子(BDNF)表达的NG108-15细胞中表达了CaM激酶II的胞浆和核同工型。转染CaM kinase II δ1和δ2后,BDNF的表达不增加,而转染CaM kinase II δ3和αB后,BDNF的表达增加。RT-PCR定量测定mRNA量的方法显示,外显子IV-BDNF mRNA增加,而外显子III-BDNF mRNA未增加。这表明核异构体只增加BDNF的mRNA和蛋白质。多巴胺D2受体(D2R)由长链受体(D2RL)和短链受体(D2RS)组成。将这两种cdna转染到NG108-15细胞中,获得了稳定表达这两种受体的细胞。激光共聚焦显微镜分析显示,细胞中每种受体的定位不同。D2R刺激增加细胞内Ca^<2+>的浓度。在稳定的D2RL细胞中瞬时转染带有核定位信号的CaM kinase II δ3 cDNA。用激动剂刺激转染细胞的D2R导致CaM激酶II核亚型的激活和BDNF表达的增加。

项目成果

期刊论文数量(160)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
M Morioka, K. Fukunaga, Y. Kai, T. Todaka, S. Yano, J. Hamada, E. Miyamoto and Y. Ushio: "Intravenously injected FK506 failed to inhibit hippocampal calcineurin"Biochem. Biophys. Res. Commun.. 286. 802-806 (2001)
M Morioka、K. Fukunaga、Y. Kai、T. Todaka、S. Yano、J. Hamada、E. Miyamoto 和 Y. Ushio:“静脉注射 FK506 未能抑制海马神经钙蛋白”Biochem。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
J.Kasahara: "Activation of Ca^<2+>/calmodulin-dependent protein kinase IV in cultured rat hippocampal neurons"J. Neurosci. Res.. 59. 594-600 (2000)
J.Kasahara:“培养的大鼠海马神经元中Ca^2/钙调蛋白依赖性蛋白激酶IV的激活”J。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
宮本英七: "プロテインキナーゼ阻害薬からの開発"蛋白質核酸酵素,増刊「最先端創薬-戦略的アプローチと先端的医薬品」. 45. 845-851 (2000)
Eishichi Miyamoto:“蛋白激酶抑制剂的发展”蛋白质核酸酶,特别版“尖端药物发现 - 战略方法和先进药物”45。845-851(2000)。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
K. Fukunaga, M. Ohmitsu, E. Miyamoto, T. Sato, M. Sugimura, T. Uchida and Y. Shirasaki: "Inhibition of neuronal nitric oxide synthase activity by 3-[2-[4-(3-chloro 2-methylphenyl)-1-piperazinyl]ethyl]-5, 6-dimethoxy- l -(4-imidazolylmethyl)-1H- indazole d
K. Fukunaga、M. Ohmitsu、E. Miyamoto、T. Sato、M. Sugimura、T. Uchida 和 Y. Shirasaki:“3-[2-[4-(3-氯 2) 抑制神经元一氧化氮合酶活性”
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
N. Inagaki, M. Nishizawa, N. Arimura, H. Yamamoto, Y. Takeuchi, E. Miyamoto, K. Kaibuchi and M. Inagaki: "Activation of Ca^<2+>/calmodulin-dependent protein kinase II within post-synaptic dendritic spines of cultured hippocampal neurons"J. Biol. Chem.. 27
N. Inagaki、M. Nishizawa、N. Arimura、H. Yamamoto、Y. Takeuchi、E. Miyamoto、K. Kaibuchi 和 M. Inagaki:“Ca^2/钙调蛋白依赖性蛋白激酶 II 的激活
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

MIYAMOTO Eishichi其他文献

MIYAMOTO Eishichi的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('MIYAMOTO Eishichi', 18)}}的其他基金

Molecular and cytobiological study on regulation of synapse
突触调控的分子和细胞生物学研究
  • 批准号:
    11694295
  • 财政年份:
    1999
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
Molecular cytobiological study on hippocampal LTP and LTD
海马LTP和LTD的分子细胞生物学研究
  • 批准号:
    09044324
  • 财政年份:
    1997
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Grant-in-Aid for international Scientific Research
Molecular cytobiological study on CaィイD12+ィエD1 signaling in the cells with cultured cells
培养细胞中CaiD12+CaiD1信号传导的分子细胞生物学研究
  • 批准号:
    09480223
  • 财政年份:
    1997
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Study on the mechanism of brain plasticity
脑可塑性机制研究
  • 批准号:
    07044281
  • 财政年份:
    1995
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Grant-in-Aid for international Scientific Research
Intracellular responses by stimulation of receptors in neurons and related cells
通过刺激神经元和相关细胞中的受体而产生细胞内反应
  • 批准号:
    07458205
  • 财政年份:
    1995
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Establishment of stable cells by induction of cDNAs of functional proteins and preparation of models for evaluation of drug efficacy for creation of new drugs
通过功能蛋白cDNA诱导建立稳定细胞并制备用于新药研发的药效评价模型
  • 批准号:
    07557195
  • 财政年份:
    1995
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Molecular and Cellular Biological Study on the Actions of Intracellular Calcium Ion in Cultured Cells
培养细胞内钙离子作用的分子和细胞生物学研究
  • 批准号:
    05454668
  • 财政年份:
    1993
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)
Study on long-term potentiation of brain hippocampus
大脑海马长时程增强研究
  • 批准号:
    04044134
  • 财政年份:
    1992
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Grant-in-Aid for international Scientific Research
The Cytophamacological Study on the Responses of the Receptors in the Cell System
细胞系统受体反应的细胞病理学研究
  • 批准号:
    02454139
  • 财政年份:
    1990
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)

相似海外基金

CAREER: Elucidating spatial and epigenetic regulation of gene expression during human development using photopatterning and single-cell multiomics
职业:利用光模式和单细胞多组学阐明人类发育过程中基因表达的空间和表观遗传调控
  • 批准号:
    2339849
  • 财政年份:
    2024
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Continuing Grant
CAREER: Scalable algorithms for regularized and non-linear genetic models of gene expression
职业:基因表达的正则化和非线性遗传模型的可扩展算法
  • 批准号:
    2336469
  • 财政年份:
    2024
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Continuing Grant
CAREER: Epigenetic Regulation of Gene Expression in Engineered Prokaryotes
职业:工程原核生物基因表达的表观遗传调控
  • 批准号:
    2338573
  • 财政年份:
    2024
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Continuing Grant
MFB: RNA modifications of frameshifting stimulators: cellular platforms to engineer gene expression by computational mutation predictions and functional experiments
MFB:移码刺激器的RNA修饰:通过计算突变预测和功能实验来设计基因表达的细胞平台
  • 批准号:
    2330628
  • 财政年份:
    2024
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Standard Grant
22-BBSRC/NSF-BIO Building synthetic regulatory units to understand the complexity of mammalian gene expression
22-BBSRC/NSF-BIO 构建合成调控单元以了解哺乳动物基因表达的复杂性
  • 批准号:
    BB/Y008898/1
  • 财政年份:
    2024
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Research Grant
How does the chromatin remodeller CHD4 regulate gene expression?
染色质重塑因子 CHD4 如何调节基因表达?
  • 批准号:
    DP240102119
  • 财政年份:
    2024
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Discovery Projects
Data-driven model links BMIz to gene expression in pediatric asthma
数据驱动模型将 BMIz 与小儿哮喘基因表达联系起来
  • 批准号:
    493135
  • 财政年份:
    2023
  • 资助金额:
    $ 8.58万
  • 项目类别:
Regulation of gene expression by the La and La-related proteins
La 和 La 相关蛋白对基因表达的调节
  • 批准号:
    489704
  • 财政年份:
    2023
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Operating Grants
Investigating the role of SARS-CoV-2 and MERS-CoV transcription regulatory sequence (TRS) in viral gene expression and virulence
研究 SARS-CoV-2 和 MERS-CoV 转录调控序列 (TRS) 在病毒基因表达和毒力中的作用
  • 批准号:
    494272
  • 财政年份:
    2023
  • 资助金额:
    $ 8.58万
  • 项目类别:
    Operating Grants
Application for 2024 CIHR NIF (ECR): Investigating the role of SARS-CoV-2 and MERS-CoV transcription regulatory sequence (TRS) in viral gene expression and virulence
2024 CIHR NIF (ECR) 申请:研究 SARS-CoV-2 和 MERS-CoV 转录调控序列 (TRS) 在病毒基因表达和毒力中的作用
  • 批准号:
    491942
  • 财政年份:
    2023
  • 资助金额:
    $ 8.58万
  • 项目类别:
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了