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

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 o...

Full description

Saved in:
Bibliographic Details
Published in:BMC molecular biology Vol. 10; no. 1; p. 88
Main Authors: Kobolak, Julianna, Kiss, Katalin, Polgar, Zsuzsanna, Mamo, Solomon, Rogel-Gaillard, Claire, Tancos, Zsuzsanna, Bock, Istvan, Baji, Arpad G, Tar, Krisztina, Pirity, Melinda K, Dinnyes, Andras
Format: Journal Article
Language:English
Published: England BioMed Central Ltd 04-09-2009
BioMed Central
Subjects:
Agricultural biotechnology
Analysis
Animal models in research
Animals
Base Sequence
Binding sites
Blastocyst - metabolism
Cloning
Embryonic Development
Embryos
Experiments
Female
Gene expression
Gene Expression Regulation, Developmental
Genetic aspects
Genetic transcription
Genetics
Humans
Life Sciences
Molecular Sequence Data
Octamer Transcription Factor-3 - genetics
Octamer Transcription Factor-3 - metabolism
Promoter Regions, Genetic
Rabbits - embryology
Rabbits - genetics
Rabbits - metabolism
Sequence Homology, Nucleic Acid
Studies
transcription factor
pseudogene
oocyte
rabbit
messenger-rna
pluripotency
pou5f1 gene
nuclear transfer
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1471-2199
1471-2199
DOI:10.1186/1471-2199-10-88