Characterization of transgenic mice constitutively expressing dominant negative form of BMAL1.

组成性表达 BMAL1 显性失活形式的转基因小鼠的表征。

基本信息

批准号：
16590069
负责人：
SHIMBA Shigeki
金额：
$ 1.98万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590069/
关键词：
BMAL1 Adipocytes metabolic syndrome 体内時計トランスジェニックマウス

项目摘要

BMAL1, also termed as MOP3 or Arnt3, is a transcription factor known to regulate circadian rhythm. Here, we established its involvement in the control of adipogenesis and lipid metabolism activity in mature adipocytes. During adipose differentiation in 3T3-L1 cells, the level of BMAL1 mRNA began to increase 4 days after induction and was highly expressed in differentiated cells. In white adipose tissues isolated from C57BL/6J mice, BMAL1 was predominantly expressed in a fraction containing adipocytes, as compared with the stromal-vascular fraction. BMAL1 knockout mice embryonic fibroblast cells (MEFs) failed to be differentiated into adipocytes. Importantly, adding BMAL1 back by adenovirus gene transfer restored the ability of BMAL1 knockout MEFs to differentiate. Knock-down of BMAL1 expression in 3T3-L1 cells by an RNAi technique allowed the cells to accumulate only minimum amounts of lipid droplets in the cells. Adenovirus-mediated expression of BMAL1 in 3T3-L1 adipocytes resulted in induction of several factors involved in lipogenesis. The promoter activity of these genes was stimulated in a BMAL1-dependent manner. Interestingly, expression of these factors showed clear circadian rhythm in mice adipose tissue. Also, overexpression of BMAL1 in adipocytes increased lipid synthesis activity. To understand the roles of BMAL1 in adipocytes in vivo, we established transgenic mice constitutively expressing BMAL1 in adipocytes under the control of adiponectin promoter. These mice will be the useful tools to analyze the function of BMAL1 in mature adipocytes. These results indicate that BMAL1, a master regulator of circadian rhythm, plays important roles also in the regulation of adipose differentiation and lipogenesis in mature adipocytes.

BMAL1（也称为MOP3或ARNT3）是已知调节昼夜节律的转录因子。在这里，我们确定了它参与成熟脂肪细胞中脂肪生成和脂质代谢活性的控制。在3T3-L1细胞中的脂肪分化过程中，诱导后4天BMAL1 mRNA的水平开始升高，并在分化细胞中高度表达。在从C57BL/6J小鼠中分离的白色脂肪组织中，与基质 - 血管分数相比，BMAL1主要在含有脂肪细胞的馏分中表达。 BMAL1敲除小鼠胚胎成纤维细胞（MEF）未能分化为脂肪细胞。重要的是，通过腺病毒基因转移添加BMAL1恢复了BMAL1基因敲除MEF分化的能力。通过RNAi技术在3T3-L1细胞中敲低BMAL1表达，使细胞仅积聚细胞中的最小脂质液滴。腺病毒介导的BMAL1在3T3-L1脂肪细胞中的表达导致诱导脂肪生成的几个因素。这些基因的启动子活性以BMAL1依赖性方式刺激。有趣的是，这些因素的表达显示小鼠脂肪组织中的昼夜节律清晰。同样，脂肪细胞中BMAL1的过表达增加了脂质合成活性。为了了解BMAL1在体内的脂肪细胞中的作用，我们在脂联素启动子控制下建立了转基因小鼠在脂肪细胞中组成型表达BMAL1。这些小鼠将是分析BMAL1在成熟脂肪细胞中的功能的有用工具。这些结果表明，昼夜节律的主要调节剂BMAL1在调节成熟脂肪细胞中的脂肪分化和脂肪形成方面也起着重要作用。

项目成果

期刊论文数量（22）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

BMAL1, a component of the molecular clock, regulates adipogenesis.

BMAL1 是分子钟的一个组成部分，调节脂肪生成。

DOI：
发表时间：
2005
期刊：
Proc. Natl. Acad. Sci. U.S.A. 102
影响因子：
0
作者：
Shimba;S.;Ishii;N.;Ohta;Y.;Ohno;T;Watabe;Y.;Hayashi;M.;Wada;T.;Aoyagi;T.;Tezuka;M
通讯作者：
M

Characteristics of circadian gene expressions in mice white adipose tissure and 3T3-L1 adipocytes.

小鼠白色脂肪组织和3T3-L1脂肪细胞昼夜节律基因表达特征。