Observation of mechanisms for transcriptional regulation

转录调控机制的观察

• 批准号: 13670031
• 负责人: WATANABE Jun
• 金额: $ 2.24万
• 依托单位: Kansai Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670031/
• 关键词: arylhydrocarbon receptor; HSP90; transcriptional regulation; CYP1A1; CYP2B2; Neural tissues; liver; rat; arylhydrocarbon receptor; 細胞内輸送; メチルコランスレン; 脳

1.By the chemiluminescent or fluorescent immunohistochemistry, HSP90 localized with arylhydrocarbon receptor(AhR) was dissociated from AhR in the cytoplasm but not in the nuclei after the stimulation of 3 -methylcholanthrene in the H4IIE cell lines and hepatocytes in sections from rat livers. However, FLET analysis revealed that the both proteins associated with a loose connection or with interconnecting factor(s). We applied this technique to examine the intracellular localization of a novel protein that localized exclusively in the endocrine cells.2.Aclose relationship was found between the expression of CYP2B2 and area of endoplasmic reticulum(ER) in hepatocytes from normal and phenobarbital-treated rats as revealed by a combination of highly-sensitive and quantitative in situ hybridization of CYP2B2 mRNA with stereological analysis of ER.3.When a reporter gene was connected to 5'-region of CYP1A1 gene, we could detect the localization of the gene by the photon imaging. However, no significant difference was found in the DNA structure if 5 '-region of CYP1A1 gene and AhR-Arnt were reacted under the presence of transcriptional factors, Sp1 or CBP/p-300 by the photon detection and/or atomic-force microscopy.

1.化学发光或荧光免疫组化显示，H4IIE细胞系和大鼠肝脏切片中的肝细胞经3-甲基胆蒽刺激后，定位于芳基烃受体(AhR)的HSP90在细胞质中与AhR解离，但不在细胞核中解离。然而，FLET 分析表明，这两种蛋白质都与松散连接或互连因子相关。我们应用该技术来检查一种仅定位于内分泌细胞的新型蛋白质的细胞内定位。 2.通过CYP2B2 mRNA与CYP2B2 mRNA的高灵敏和定量原位杂交相结合，发现正常和苯巴比妥治疗的大鼠肝细胞中CYP2B2的表达与内质网（ER）面积之间存在密切关系。 ER的体视学分析。3.当CYP1A1基因的5'区连接报告基因时，可以通过光子成像检测该基因的定位。然而，通过光子检测和/或原子力显微镜，在转录因子Sp1或CBP/p-300存在下，CYP1A1基因的5'区与AhR-Arnt反应，DNA结构没有发现显着差异。

项目成果

Y.Utsunomiya, J.Watanabe, Y.Takamori, Y.Kataoka, K.Kurokawa, H.Yamada: "Crossreaction with an anti-Bax antibody reveales nobel multi-endocrine cellular antigen."J.Histochem.Cytochem.. 52. 805-812 (2004)

Y.Utsunomiya、J.Watanabe、Y.Takamori、Y.Kataoka、K.Kurokawa、H.Yamada：“与抗 Bax 抗体的交叉反应揭示了诺贝尔多内分泌细胞抗原。”J.Histochem.Cytochem.. 52。

Y.Cui, Y.Kataoka, Q.H.Li, C.Yokoyama, A.Yamagata, N.Mochizuki-Oda, J.Watanabe, H.Yamada, Y.Watanabe: "Targeted tissue oxidation in the cerebral cortex induces local prolonged depolarization and cortical spreading depression in the rat brain"Biochem Biophy

Y.Cui、Y.Kataoka、Q.H.Li、C.Yokoyama、A.Yamagata、N.Mochizuki-Oda、J.Watanabe、H.Yamada、Y.Watanabe：“大脑皮层的目标组织氧化会导致局部长时间去极化，

Y.Cui, Y.Kataoka, Q.H.Li, C.Yokoyama, A.Yamagata, N.Mochizuki-Oda, J.Watanabe, et al.: "Targeted tissue oxidation in the cerebral cortex induces local prolonged depolarization and cortical spreading depression in the rat brain"Biochem.Biophys.Res.Commun..

Y.Cui、Y.Kataoka、Q.H.Li、C.Yokoyama、A.Yamagata、N.Mochizuki-Oda、J.Watanabe 等人：“大脑皮层中的目标组织氧化会导致局部长期去极化和皮层扩散抑制

Utsunomiya Y: "Crossreaction with an anri-Bax antibody reveales nobel multi-endocrine cellular antigen."J.Histochem.Cytochem.. 52(印刷中). (2004)

Utsunomiya Y：“与 anri-Bax 抗体的交叉反应揭示了诺贝尔多内分泌细胞抗原。”J.Histochem.Cytochem.. 52（印刷中）。

Utsunomiya Y, Watanabe J, Takamori Y, et al.: "Crossreaction with an anti-Bax antibody reveals novel multi-endocrine cellular antigen."J.Histochem.Cytochem.. 52. 805-812 (2004)

Utsunomiya Y、Watanabe J、Takamori Y 等人：“与抗 Bax 抗体的交叉反应揭示了新型多内分泌细胞抗原。”J.Histochem.Cytochem.. 52. 805-812 (2004)