Promoter analysis of the rabbit POU5F1 gene and its expression in preimplantation stage embryos.
Promoter analysis of the rabbit POU5F1 gene and its expression in preimplantation stage embryos.
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DOI：
10.1186/1471-2199-10-88
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发表时间：
2009-09-04
影响因子：
--
通讯作者：
Dinnyes A
中科院分区：
文献类型：
--
作者：
Kobolak J;Kiss K;Polgar Z;Mamo S;Rogel-Gaillard C;Tancos Z;Bock I;Baji AG;Tar K;Pirity MK;Dinnyes A
POU5F1基因编码八聚体结合转录因子-4 （Oct4）。它在胚胎发育过程中对多能性的调控起着至关重要的作用，被广泛用作胚胎干细胞的分子标记。本研究的目的是鉴定和分析兔POU5F1基因的启动子区域；进一步研究其在着床前期兔胚胎中的表达规律。对兔POU5F1上游区域进行亚克隆测序，鉴定出4个高度保守的启动子区域（CR1-4）。兔与人的保守结构域在序列水平上的相似性最高。近端增强子区PE-1A的同源性最高（96.4%）。此外，含有远端增强子（DE-2A）的CR4调节结构域负责干细胞特异性表达。BAC文库筛选还发现了一个加工过的兔POU5F1假基因。实时荧光定量PCR实验结果显示，在卵母细胞和受精卵中，POU5F1 mRNA含量丰富，但在胚胎基因组激活后，POU5F1 mRNA含量逐渐减少，此后POU5F1 mRNA水平持续升高，直至囊胚期。利用XYClone激光系统分别对胚胎内细胞团（ICM）和滋养层进行显微解剖和检测，并在两种细胞类型中检测到POU5F1 mRNA。本研究对家兔POU5F1基因的调控区进行了比较序列分析。我们的数据表明，在哺乳动物早期发育过程中，POU5F1基因受到严格调控。我们提出含有DE-2A增强子的保守的CR4区域负责高度保守的ESC特异性基因表达。值得注意的是，我们首次报道了兔POU5F1不仅局限于ICM细胞，而且也在滋养细胞中表达。这一信息为进一步研究POU5F1基因的系统发育作用和调控机制提供了理论依据。
The POU5F1 gene encodes the octamer-binding transcription factor-4 (Oct4). It is crucial in the regulation of pluripotency during embryonic development and widely used as molecular marker of embryonic stem cells (ESCs). The objective of this study was to identify and to analyse the promoter region of rabbit POU5F1 gene; furthermore to examine its expression pattern in preimplantation stage rabbit embryos. The upstream region of rabbit POU5F1 was subcloned sequenced and four highly conserved promoter regions (CR1-4) were identified. The highest degree of similarity on sequence level was found among the conserved domains between rabbit and human. Among the enhancers the proximal enhancer region (PE-1A) exhibited the highest degree of homology (96.4%). Furthermore, the CR4 regulator domain containing the distal enhancer (DE-2A) was responsible for stem cell-specific expression. Also, BAC library screen revealed the existence of a processed pseudogene of rabbit POU5F1. The results of quantitative real-time PCR experiments showed that POU5F1 mRNA was abundantly present in oocytes and zygotes, but it was gradually reduced until the activation of the embryonic genome, thereafter a continuous increase in POU5F1 mRNA level was observed until blastocyst stage. By using the XYClone laser system the inner cell mass (ICM) and trophoblast portions of embryos were microdissected and examined separately and POU5F1 mRNA was detected in both cell types. In this study we provide a comparative sequence analysis of the regulatory region of rabbit POU5F1 gene. Our data suggest that the POU5F1 gene is strictly regulated during early mammalian development. We proposed that the well conserved CR4 region containing the DE-2A enhancer is responsible for the highly conserved ESC specific gene expression. Notably, we are the first to report that the rabbit POU5F1 is not restricted to ICM cells only, but it is expressed in trophoblast cells as well. This information may be well applicable to investigate further the possible phylogenetic role and the regulation of POU5F1 gene.
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影响因子：
4
作者：
Cauffman, G;Van de Velde, H;Van Steirteghem, A
通讯作者：
Van Steirteghem, A
影响因子：
3.6
作者：
Kurosaka, S;Eckardt, S;McLaughlin, KJ
通讯作者：
McLaughlin, KJ
DOI：
10.1089/clo.2008.0072
发表时间：
2009-03-01
期刊：
CLONING AND STEM CELLS
影响因子：
--
作者：
Meng, Qinggang;Polgar, Zsuzsanna;Dinnyes, Andras
通讯作者：
Dinnyes, Andras
影响因子：
3.6
作者：
Mitalipov, SM;Kuo, HC;Wolf, DP
通讯作者：
Wolf, DP
影响因子：
5.3
作者：
Chew, JL;Loh, YH;Ng, HH
通讯作者：
Ng, HH