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Functional analysis of proteins belongs to SIBLING family in physiological and pathological calcification

SIBLING家族蛋白在生理和病理钙化中的功能分析

基本信息

批准号：
18590371
负责人：
NOMURA Shintaro
金额：
$ 2.53万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590371/
关键词：
ostemontin mechanical stress estrogen transgenic mice cell attachment knockout mice SIBLING luciferase 石灰化骨、軟骨組織腎臓結石 RGD シュウ酸カルシウム

项目摘要

1. Identification of promoter region of Osteopontin in response to environmental change.Up-regulation of osteopontin was observed in transgenic mice carrying GFP cDNA ligated with distal promoter (-5505 bp to mRNA start site) (GFP5505 OPN) when mechanical stress was loaded to bone. Similar effect was observed in the ovaryextomized transgenic mice. Further investigation revealed that mechanical stress response element and estrogen respose element lies between -3156 and -5505 in the start site of transcription.2. Analysis of primary cultured cells derived from transgenic mice.Bone cells containing osteoblasts and osteocytes of the transgenic mice were cultured. Expression level of GFP was elevated when mechanical stress was loaded to cultured cells. Identification of mechanical stress-response element was performed by luciferase assay using various region of promoter region joined to luciferase cDNA. Finally, we identified the mechanical stress-response element between-3983 and -315G to the mRNA start site of osteopontin gene.3. Effects of truncated osteopontin on development of bone tissue.Transgenic mice carrying either calcium binding site or cell attachment site of osteopontin controlled -5505 promoter were generated. Transgenic mice were introduced to knockout mice of osteopontin gene. Generated mice showed abnormal structure in bone. The result indicate that calcium binding site and cell attachment site of osteopontin functioning in vivo.

1.骨桥蛋白启动子区域对环境变化的响应：在机械应力作用下，携带GFP基因启动子的转基因小鼠(GFP5505OPN)骨桥蛋白表达上调。在摘除卵巢的转基因小鼠中也观察到了类似的效果。进一步研究发现，在转录起始点，机械应激反应元件和雌激素反应元件位于-3156和-5505之间。转基因小鼠成骨细胞和转基因小鼠骨细胞的原代培养。当机械应力作用于培养细胞时，GFP的表达水平升高。利用荧光素酶基因启动子区的不同区域，通过荧光素酶分析鉴定机械应激反应元件。最后，我们确定了骨桥蛋白基因的-3983~-315G之间的机械应力反应元件。截短型骨桥蛋白对骨组织发育的影响：建立了携带骨桥蛋白控制的-5505启动子的钙结合部位或细胞附着部位的转基因小鼠。将转基因小鼠引入骨桥蛋白基因敲除小鼠。产生的小鼠骨骼结构异常。结果表明，骨桥蛋白的钙结合部位和细胞附着部位在体内发挥作用。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Saito-Ohara F, Inazawa J. Kohno T. Yokota J. Ishii S. Reduced levels of ATF-2 predispose mice to mammary tumors

Saito-Ohara F、Inazawa J. Kohno T. Yokota J. Ishii S. ATF-2 水平降低使小鼠易患乳腺肿瘤

DOI：
发表时间：
2007
期刊：
Mol Cell Biol 27
影响因子：
0
作者：
Maekawa;T.;Shinagawa;T.;Sano;Y.;Sakuma;T.;Nomura;S.;Nagasaki;K.;Miki;Y
通讯作者：
Y

Successful formation of calcium oxalate stone in mouse kidney by intra abdominal glyoxylate injection

腹腔注射乙醛酸盐成功形成小鼠肾草酸钙结石

DOI：
发表时间：
2007
期刊：
Urological Research 35
影响因子：
0
作者：
Ogata N;Kawaguchi H;Chung UI;Roth SI;Segre GV.;Atsushi Okada
通讯作者：
Atsushi Okada

Reduced levels of ATF-2mice to Mammary tumors

降低 ATF-2 小鼠乳腺肿瘤的水平

DOI：
发表时间：
期刊：
Mol. Cell Biol. (印刷中)
影响因子：
0
作者：
Maekawa T;Nomura S. et al.
通讯作者：
Nomura S. et al.

IL-6 blockade inhibits the induction of myelin antigen-specific Th17 cells and Th1 cells in experimental autoimmune encephalomyelitis

DOI：
10.1073/pnas.0802218105
发表时间：
2008-07-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
影响因子：
11.1
作者：
Serada, Satoshi;Fujimoto, Minoru;Naka, Tetsuji
通讯作者：
Naka, Tetsuji

Successful formation of calcium oxalate crystal deposition in mouse kidney by intraabdominal glyoxylate injection