Afficher la couverture 'Promoter analysis of the rabbit POU5F1 gene and its expression in preimplantation stage embryos | Kobolak, Julianna' en grand format
/5

0 avis

Promoter analysis of the rabbit POU5F1 gene and its expression in preimplantation stage embryos

Archive ouverte

Kobolak, Julianna | Kiss, Katalin | Polgar, Zsuzsanna | Mamo, Solomon | Gaillard, Claire | Tancos, Zsuzsanna | Bock, Istvan | Baji, Arpad | Tar, Krisztina | Pirity, Melinda | Dinnyes, Andras

Edité par CCSD ; BioMed Central -

Background: 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. [br/] Results: 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. [br/] Conclusion: 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.

Suggestions

Du même auteur

Mammalian embryo comparison identifies novel pluripotency genes associated with the naïve or primed state.

Archive ouverte | Bernardo, Andreia S | CCSD

International audience. During early mammalian development transient pools of pluripotent cells emerge that can be immortalised upon stem cell derivation. The pluripotent state, "naïve" or "primed", depends on the e...

Allele-specific RNA-seq expression profiling of imprinted genes in mouse isogenic pluripotent states

Archive ouverte | Dirks, René a M | CCSD

International audience. BACKGROUND: Genomic imprinting, resulting in parent-of-origin specific gene expression, plays a critical role in mammalian development. Here, we apply allele-specific RNA-seq on isogenic B6D2...

Enhanced cardiac differentiation of mouse embryonic stem cells by use of the slow-turning, lateral vessel (STLV) bioreactor

Archive ouverte | Rungarunlert, Sasitorn | CCSD

International audience. Embryoid body (EB) formation is a common intermediate during in vitro differentiation of pluripotent stem cells into specialized cell types. We have optimized the slow-turning, lateral vessel...

Chargement des enrichissements...