Identification of a regulatory region which is involved in Sox-2 expression in embryonic and neural stem cells and its molecular basis

胚胎和神经干细胞中参与Sox-2表达的调控区的鉴定及其分子基础

基本信息

批准号：
15590253
负责人：
OKUDA Akihiko
金额：
$ 2.18万
依托单位：
Saitama Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

We have been interested in understanding common properties between ES cells and neural stem cells such as self-renewal ability in a molecular level. To gain insight about it, we considered that it is important to identify a regulatory region which works in both of these two kinds of stem cells. Since it is known that Sox-2 gene is expressed in both ES cells and neural stem cells, we pursued the possibility that the Sox-2 gene bears such regulatory region. We have previously identified two enhancers, SRR1 and SRR2 based on their activities in pluripotent ES cells. In this context, we examined whether these Sox-2 regulatory regions exhibit their activities also in neural stem/progenitor cells. Indeed, experiments with nestin-positive cells derived from ES cells, neurosphere methods, and in utero electroporation in all cases revealed that both of SRR1 and SRR2 exert rather strong transcription stimulating activities in neural stem/progenitor cells as well as in ES cells. Moreover, as in the case of ES cells, activities of SRR1 and SRR2 in neural stem/progenitor cells are specific in their multipotent state and these enhancers lose their entire activities when neural stem/progenitor cells are induced to differentiate with serum. Mutagenesis analyses demonstrated that SRR2 exhibits its activities in these two different types of stem cells by utilizing a common core sequence. Our analyses also revealed that Brn-1-Sox-2 and Brn-2-Sox-2 complexes are involved in SRR2 activities in neural cells, while Oct-3/4-Sox-2 complex plays major role in ES cells.We also performed the transgenic analyses about the SRR2 enhancer. From these analyses, we found that the SRR2 do not function in neural stem/progenitor cells in general, but exerts its activity rather specifically in neural stem/progenitor cells residing in telencephalon.

我们一直对了解 ES 细胞和神经干细胞之间的共同特性感兴趣，例如分子水平上的自我更新能力。为了深入了解这一点，我们认为确定在这两种干细胞中都起作用的调节区域非常重要。由于已知Sox-2基因在ES细胞和神经干细胞中均表达，因此我们探究Sox-2基因具有这样的调控区域的可能性。我们之前根据 SRR1 和 SRR2 在多能 ES 细胞中的活性鉴定了两种增强子。在这种情况下，我们检查了这些 Sox-2 调节区是否也在神经干/祖细胞中表现出其活性。事实上，在所有情况下对来源于 ES 细胞的巢蛋白阳性细胞、神经球方法和子宫内电穿孔进行的实验表明，SRR1 和 SRR2 在神经干/祖细胞以及 ES 细胞中都发挥相当强的转录刺激活性。此外，与ES细胞的情况一样，神经干/祖细胞中SRR1和SRR2的活性在其多能状态下是特异的，并且当神经干/祖细胞被血清诱导分化时，这些增强子失去其全部活性。诱变分析表明，SRR2 通过利用共同的核心序列在这两种不同类型的干细胞中表现出其活性。我们的分析还表明，Brn-1-Sox-2 和 Brn-2-Sox-2 复合物参与神经细胞中的 SRR2 活性，而 Oct-3/4-Sox-2 复合物在 ES 细胞中起主要作用。进行了有关SRR2增强子的转基因分析。从这些分析中，我们发现SRR2一般不在神经干/祖细胞中发挥作用，而是在端脑中的神经干/祖细胞中相当特异地发挥其活性。

项目成果

期刊论文数量（24）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

「研究成果報告書概要(欧文)」より

摘自《研究结果报告摘要（欧洲）》

DOI：
发表时间：
2006
期刊：
Seibutsu Butsuri 46(1)
影响因子：
0
作者：
Yasushi Shigeri;Keiko Shimamoto
通讯作者：
Keiko Shimamoto

Oct-3/4 maintains the proliferative ES cell state via specific binding to a variant octamer sequence in the regulatory region of the UTF1 locus

Oct-3/4 通过与 UTF1 基因座调控区的变异八聚体序列特异性结合来维持 ES 细胞增殖状态

DOI：
发表时间：
期刊：
Molecular and Cellular Biology (in press)
影响因子：
0
作者：
M.Narita;J.Khotib;H.Mizuguchi;M.Suzuki;S.Ozaki;Y.Yajima;L.F.Tseng;T.Suzuki;Masazumi Nishimoto
通讯作者：
Masazumi Nishimoto

Masazumi Nishimoto: "The embryonic Octamer factor 3/4 displays distinct DNA binding specificity from those of other Octamer factors"Biochem.Biophys.Res.Commun.. 302. 581-586 (2003)

Masazumi Nishimoto：“胚胎八聚体因子 3/4 显示出与其他八聚体因子不同的 DNA 结合特异性”Biochem.Biophys.Res.Commun.. 302. 581-586 (2003)

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Satoru Miyagi: "The Sox-2 regulatory regions display their activities in two distinct multipotent stem cells"Mol.Cell.Biol.. (in press). (2004)

Satoru Miyagi：“Sox-2 调节区在两种不同的多能干细胞中显示出它们的活性”Mol.Cell.Biol..（正在出版）。

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Expression of tbx20 RNA during chick heart development

DOI：
10.1002/dvdy.20076
发表时间：
2004-07
期刊：
Developmental Dynamics
影响因子：
2.5
作者：
T. Yamagishi;Y. Nakajima;S. Nishimatsu;T. Nohno;Katsumi Ando;Hiroaki Nakamura
通讯作者：
T. Yamagishi;Y. Nakajima;S. Nishimatsu;T. Nohno;Katsumi Ando;Hiroaki Nakamura