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

Julianna Kobolak, Katalin Kiss, Zsuzsanna Polgar, Solomon Mamo, Claire Rogel-Gaillard, Zsuzsanna Tancos, Istvan Bock, Arpad G. Baji, Krisztina Tar, M. Pirity, A. Dinnyés

Research output: Contribution to journalArticle

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Abstract

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. 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. 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.

Original languageEnglish
Article number1471
Pages (from-to)88
Number of pages1
JournalBMC Molecular Biology
Volume10
DOIs
Publication statusPublished - Sep 4 2009

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Blastocyst
Rabbits
Gene Expression
Genes
Trophoblasts
Embryonic Stem Cells
Genetic Promoter Regions
Messenger RNA
Octamer Transcription Factors
Embryonic Structures
Pseudogenes
Zygote
Nucleic Acid Regulatory Sequences
Libraries
Oocytes
Embryonic Development
Sequence Analysis
Real-Time Polymerase Chain Reaction
Lasers
Stem Cells

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Promoter analysis of the rabbit POU5F1 gene and its expression in preimplantation stage embryos. / Kobolak, Julianna; Kiss, Katalin; Polgar, Zsuzsanna; Mamo, Solomon; Rogel-Gaillard, Claire; Tancos, Zsuzsanna; Bock, Istvan; Baji, Arpad G.; Tar, Krisztina; Pirity, M.; Dinnyés, A.

In: BMC Molecular Biology, Vol. 10, 1471, 04.09.2009, p. 88.

Research output: Contribution to journalArticle

Kobolak, J, Kiss, K, Polgar, Z, Mamo, S, Rogel-Gaillard, C, Tancos, Z, Bock, I, Baji, AG, Tar, K, Pirity, M & Dinnyés, A 2009, 'Promoter analysis of the rabbit POU5F1 gene and its expression in preimplantation stage embryos', BMC Molecular Biology, vol. 10, 1471, pp. 88. https://doi.org/10.1186/1471-2199-10-88
Kobolak, Julianna ; Kiss, Katalin ; Polgar, Zsuzsanna ; Mamo, Solomon ; Rogel-Gaillard, Claire ; Tancos, Zsuzsanna ; Bock, Istvan ; Baji, Arpad G. ; Tar, Krisztina ; Pirity, M. ; Dinnyés, A. / Promoter analysis of the rabbit POU5F1 gene and its expression in preimplantation stage embryos. In: BMC Molecular Biology. 2009 ; Vol. 10. pp. 88.
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AU - Mamo, Solomon

AU - Rogel-Gaillard, Claire

AU - Tancos, Zsuzsanna

AU - Bock, Istvan

AU - Baji, Arpad G.

AU - Tar, Krisztina

AU - Pirity, M.

AU - Dinnyés, A.

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