Differential Transcriptional Regulation of the NANOG Gene in Chicken Primordial Germ Cells and Embryonic Stem Cells

Abstract BackgroundNANOG is a core transcription factor (TF) in embryonic stem cells (ESCs) and primordial germ cells (PGCs). Regulation of the NANOG gene by TFs, epigenetic factors, and autoregulatory factors is well characterized in ESCs, and transcriptional regulation of NANOG is well established in these cells. Although NANOG plays a key role in germ cells, the molecular mechanism underlying its transcriptional regulation in PGCs has not been studied. Therefore, we investigated the mechanism that regulates transcription of the chicken NANOG (cNANOG) gene in PGCs and ESCs. ResultsWe first identified the transcription start site of cNANOG by 5’-rapid amplification of cDNA ends PCR analysis. Then, we measured the promoter activity of various 5’ flanking regions of cNANOG in chicken PGCs and ESCs using the luciferase reporter assay. cNANOG expression required transcriptional cis-regulatory elements, which were positively regulated by POU5F3 (OCT4) and SOX2 and negatively regulated by TP53 in PGCs. The proximal region of the cNANOG promoter contains a positive cis-regulatory element (CCAAT/enhancer-binding protein (CEBP)-binding site) in ESCs. Furthermore, small interfering RNA-mediated knockdown demonstrated that POU5F3, SOX2, and CEBP played a role in cell type-specific transcription of cNANOG.ConclusionsWe show for the first time that different cis-regulatory elements control transcription of cNANOG in a cell type-specific manner. This finding might help to elucidate the mechanism that regulates cNANOG expression in PGCs and ESCs.

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Differential transcriptional regulation of the NANOG gene in chicken primordial germ cells and embryonic stem cells

Journal of Animal Science and Biotechnology ◽

10.1186/s40104-021-00563-5 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Hee Jung Choi ◽

So Dam Jin ◽

Deivendran Rengaraj ◽

Jin Hwa Kim ◽

Bertrand Pain ◽

...

Keyword(s):

Stem Cells ◽

Transcriptional Regulation ◽

Embryonic Stem Cells ◽

Germ Cells ◽

Primordial Germ Cells ◽

Embryonic Stem ◽

Regulatory Elements ◽

Cell Type ◽

Transcriptional Regulatory ◽

Cell Type Specific

Abstract Background NANOG is a core transcription factor (TF) in embryonic stem cells (ESCs) and primordial germ cells (PGCs). Regulation of the NANOG gene by TFs, epigenetic factors, and autoregulatory factors is well characterized in ESCs, and transcriptional regulation of NANOG is well established in these cells. Although NANOG plays a key role in germ cells, the molecular mechanism underlying its transcriptional regulation in PGCs has not been studied. Therefore, we investigated the mechanism that regulates transcription of the chicken NANOG (cNANOG) gene in PGCs and ESCs. Results We first identified the transcription start site of cNANOG by 5′-rapid amplification of cDNA ends PCR analysis. Then, we measured the promoter activity of various 5′ flanking regions of cNANOG in chicken PGCs and ESCs using the luciferase reporter assay. cNANOG expression required transcriptional regulatory elements, which were positively regulated by POU5F3 (OCT4) and SOX2 and negatively regulated by TP53 in PGCs. The proximal region of the cNANOG promoter contains a positive transcriptional regulatory element (CCAAT/enhancer-binding protein (CEBP)-binding site) in ESCs. Furthermore, small interfering RNA-mediated knockdown demonstrated that POU5F3, SOX2, and CEBP played a role in cell type-specific transcription of cNANOG. Conclusions We show for the first time that different trans-regulatory elements control transcription of cNANOG in a cell type-specific manner. This finding might help to elucidate the mechanism that regulates cNANOG expression in PGCs and ESCs.

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Collagen-alginate microspheres as a 3D culture system for mouse embryonic stem cells differentiation to primordial germ cells

Biologicals ◽

10.1016/j.biologicals.2017.04.003 ◽

2017 ◽

Vol 48 ◽

pp. 114-120 ◽

Cited By ~ 4

Author(s):

Vahid Mansouri ◽

Mohammad Salehi ◽

Mir davood Omrani ◽

Zahra Niknam ◽

Abdolreza Ardeshirylajimi

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Germ Cells ◽

Culture System ◽

Primordial Germ Cells ◽

Embryonic Stem ◽

3D Culture ◽

Mouse Embryonic Stem Cells ◽

3D Culture System

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Primordial germ cells can be differentiated by retinoic acid and progesterone induction from embryonic stem cells

Journal of Biosciences ◽

10.1007/s12038-021-00210-1 ◽

2021 ◽

Vol 46 (3) ◽

Author(s):

Meysam Hassani Moghaddam ◽

Neda Eskandari ◽

Hossein Nikzad ◽

Mohammad Miryounesi ◽

Mohammad Karimian ◽

...

Keyword(s):

Stem Cells ◽

Retinoic Acid ◽

Embryonic Stem Cells ◽

Germ Cells ◽

Primordial Germ Cells ◽

Embryonic Stem

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An Evidence That Murine Marrow-Derived CXCR4+ SSEA-1+ Oct-4+ Very Small Embryonic-Like (VSEL) Stem Cells Are Pluripotent and Express Several Primordial Germ Cell (PGC) Markers - Hypotesis for Developmental Deposition of PGC in Various Organs.

Blood ◽

10.1182/blood.v108.11.1676.1676 ◽

2006 ◽

Vol 108 (11) ◽

pp. 1676-1676 ◽

Cited By ~ 1

Author(s):

Magda Kucia ◽

Ewa Zuba-Surma ◽

Ryan Reca ◽

Janina Ratajczak ◽

Mariusz Ratajczak

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Germ Cell ◽

Germ Cells ◽

Pluripotent Stem Cells ◽

Fetal Liver ◽

Primordial Germ Cells ◽

Embryonic Stem ◽

Embryoid Bodies ◽

C2c12 Cells

Abstract Recently we identified in murine BM a homogenous population of rare (~0.01% of BMMNC) Sca-1+ lin− CD45− cells that express by RQ-PCR and immunhistochemistry markers of pluripotent stem cells (PSC) such as SSEA-1, Oct-4, Nanog and Rex-1 and highly express Rif-1 telomerase protein (Leukemia2006;20,857–869). Direct electronmicroscopical analysis revealed that these cells display several features typical for embryonic stem cells such as i) small size (2–4 um in diameter), ii) large nuclei surrounded by a narrow rim of cytoplasm, and iii) open-type chromatin (euchromatin). We also found that VSELs may be released from BM and circulate in peripheral blood during tissue/organ injuries (e.g., heart infarct, stroke). Recently we noticed that ~5–10% of purified VSELs if plated over a C2C12 murine sarcoma cell feeder layer are able to form spheres that resemble embryoid bodies. Cells from these VSEL-derived spheres (VSEL-DS) are composed of immature cells with large nuclei containing euchromatin, and similarly as purified VSELs are CXCR4+SSEA-1+Oct-4+. Furthermore, VSEL-DS after replating over C2C12 cells may again (up to 5–7 passages) grow new spheres or if plated into cultures promoting tissue differentiation expand into cells from all three germ-cell layers. The formation of VSEL-DS was observed in a presence of C2C12 cells obtained from different sources. Furthermore, VSELs isolated from GFP+ mice grew GFP+ VSEL-DS which show a diploid content of DNA. This suggests that VSEL-DS are in fact derived from VSELs and not from the supportive C2C12 cell line as well as excludes the possibility of cell fusion to the observed phenomenon. Similar spheres were also formed by VSELs isolated from murine fetal liver, spleen and thymus. Interestingly formation of VSEL-DS was associated with a young age, and no VSEL-DS were observed by cells isolated from old mice (> 2 years). We also found that cells isolated from VSEL-DS similarly as embryonic stem cells grow tumors after injection into immunodeficient NOD/SCID mice (51/52 inoculated mice). Since VSELs isolated by us express several markers of primordial germ cells (fetal-type alkaline phosphatase, Oct-4, SSEA-1, CXCR4, Mvh, Stella, Fragilis, Nobox, Hdac6) we hypothesize that VSELs are closely related to a population of primordial germ cells. These cells are specified during early gastrulation in the proximal epiblast and subsequently migrate in a CXCR4-SDF-1 dependent manner through the embryo proper to their final destination in genital ridges. It is possible that some of these cells or a population of cells closely related to them migrate astray being chemoattracted by SDF-1 to fetal liver and subsequently, during the third trimester of gestation seed together with hematopoietic stem cells in bone marrow and perhaps other organs as well. In conclusion, we postulate that VSELs identified by us and purified at the single cell level could become an important source of pluripotent stem cells for regeneration.

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