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

• 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
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590253/
• 关键词: ES cells; Neural stem cells; enhancer; undifferentiated state; pluripotency; in utero electroporation; Sox-2; octamer factor; 多能性・自己増殖性

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.

我们一直有兴趣在分子水平上了解ES细胞和神经干细胞之间的共同特性，如自我更新能力。为了深入了解这一点，我们认为重要的是要确定一个调节区域，在这两种干细胞中工作。由于已知Sox-2基因在ES细胞和神经干细胞中表达，我们继续研究Sox-2基因具有这样的调控区的可能性。我们以前已经确定了两个增强子，SRR 1和SRR 2的基础上，他们在多能ES细胞的活动。在这种情况下，我们研究了这些Sox-2调控区是否也在神经干/祖细胞中表现出它们的活性。事实上，在所有情况下，用来自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，SRR 1和SRR 2在神经干/祖细胞以及ES细胞中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SRR 1和SRR 2的活性在其多能状态下是特异性的，并且当用血清诱导神经干/祖细胞分化时，这些增强剂失去其全部活性。突变分析表明，SRR 2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复合物参与了神经细胞中SRR 2的活性，而Oct-3/4-Sox-2复合物在ES细胞中起主要作用。从这些分析中，我们发现SRR 2一般在神经干/祖细胞中不起作用，但在端脑中的神经干/祖细胞中特异性地发挥其活性。

项目成果

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

• 期刊: Molecular and Cellular Biology (in press)
• 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)

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）。

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

• DOI: 10.1128/mcb.25.12.5084-5094.2005
• 发表时间: 2005-06-01
• 期刊: MOLECULAR AND CELLULAR BIOLOGY
• 影响因子: 5.3
• 作者: Nishimoto, M;Miyagi, S;Okuda, A
• 通讯作者: Okuda, A

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