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Epigenetic Modification Agents Improve Gene-Specific Methylation Reprogramming in Porcine Cloned Embryos.

Huan Y, Wu Z, Zhang J, Zhu J, Liu Z, Song X - PLoS ONE (2015)

Bottom Line: In this study, we found that compared with in vitro fertilized counterparts, cloned embryos displayed the disrupted patterns of Oct4 demethylation and Thy1 remethylation.In conclusion, our results demonstrated that cloned embryos displayed incomplete gene-specific methylation reprogramming and disrupted expression patterns of pluripotency and tissue specific genes, and epigenetic modification agents improved gene-specific methylation reprogramming and expression pattern by regulating epigenetic modification related genes.This work would have important implications in improving cloning efficiency.

View Article: PubMed Central - PubMed

Affiliation: College of Animal Science and Technology, Qingdao Agricultural University, Qingdao, Shandong Province, China.

ABSTRACT
Incomplete DNA methylation reprogramming in cloned embryos leads to poor cloning efficiency. Epigenetic modification agents can improve genomic methylation reprogramming and the development of cloned embryos, however, the effect of epigenetic modification agents on gene-specific methylation reprogramming remains poorly studied. Here, we investigated DNA methylation reprogramming of pluripotency (Oct4) and tissue specific (Thy1) genes during early embryo development in pigs. In this study, we found that compared with in vitro fertilized counterparts, cloned embryos displayed the disrupted patterns of Oct4 demethylation and Thy1 remethylation. When 5-aza-2'-deoxycytidine (5-aza-dC) or trichostatin A (TSA) enhanced the development of cloned embryos, the transcripts of DNA methyltransferases (Dnmt1 and Dnmt3a), histone acetyltransferase 1 (Hat1) and histone deacetylase 1 (Hdac1) and the methylation and expression patterns of Oct4 and Thy1 became similar to those detected in in vitro fertilized counterparts. Further studies showed that Dnmt1 knockdown in cloned embryos enhanced the methylation reprogramming of Oct4 and Thy1 and promoted the activation of Oct4 and the silence of Thy1. In conclusion, our results demonstrated that cloned embryos displayed incomplete gene-specific methylation reprogramming and disrupted expression patterns of pluripotency and tissue specific genes, and epigenetic modification agents improved gene-specific methylation reprogramming and expression pattern by regulating epigenetic modification related genes. This work would have important implications in improving cloning efficiency.

No MeSH data available.


Related in: MedlinePlus

Relative Oct4 and Thy1 transcripts in early embryos.A and B, relative transcripts of Oct4 and Thy1 at 1-cell, 2-cell, 4-cell, 8-cell and blastocyst stages of IVF, NT-CON, NT-AZA and NT-TSA embryos, respectively. 5-aza-dC or TSA appropriately promoted the expression of Oct4 and silenced the transcription of Thy1 in cloned embryos. The transcript abundance in MII oocytes (A) or cloned embryos at 2 h post activation (B) was considered to be the control. The data were expressed as mean ± SEM. a-cValues at a given stage for the same gene with different superscripts differ significantly (P<0.05).
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pone.0129803.g003: Relative Oct4 and Thy1 transcripts in early embryos.A and B, relative transcripts of Oct4 and Thy1 at 1-cell, 2-cell, 4-cell, 8-cell and blastocyst stages of IVF, NT-CON, NT-AZA and NT-TSA embryos, respectively. 5-aza-dC or TSA appropriately promoted the expression of Oct4 and silenced the transcription of Thy1 in cloned embryos. The transcript abundance in MII oocytes (A) or cloned embryos at 2 h post activation (B) was considered to be the control. The data were expressed as mean ± SEM. a-cValues at a given stage for the same gene with different superscripts differ significantly (P<0.05).

Mentions: Generally, the improvement of DNA methylation reprogramming would effectively regulate gene expression [17]. Thus, the enhanced methylation reprogramming of Oct4 and Thy1 induced by 5-aza-dC or TSA should lead to their appropriate expression (Fig 3).


Epigenetic Modification Agents Improve Gene-Specific Methylation Reprogramming in Porcine Cloned Embryos.

Huan Y, Wu Z, Zhang J, Zhu J, Liu Z, Song X - PLoS ONE (2015)

Relative Oct4 and Thy1 transcripts in early embryos.A and B, relative transcripts of Oct4 and Thy1 at 1-cell, 2-cell, 4-cell, 8-cell and blastocyst stages of IVF, NT-CON, NT-AZA and NT-TSA embryos, respectively. 5-aza-dC or TSA appropriately promoted the expression of Oct4 and silenced the transcription of Thy1 in cloned embryos. The transcript abundance in MII oocytes (A) or cloned embryos at 2 h post activation (B) was considered to be the control. The data were expressed as mean ± SEM. a-cValues at a given stage for the same gene with different superscripts differ significantly (P<0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4465902&req=5

pone.0129803.g003: Relative Oct4 and Thy1 transcripts in early embryos.A and B, relative transcripts of Oct4 and Thy1 at 1-cell, 2-cell, 4-cell, 8-cell and blastocyst stages of IVF, NT-CON, NT-AZA and NT-TSA embryos, respectively. 5-aza-dC or TSA appropriately promoted the expression of Oct4 and silenced the transcription of Thy1 in cloned embryos. The transcript abundance in MII oocytes (A) or cloned embryos at 2 h post activation (B) was considered to be the control. The data were expressed as mean ± SEM. a-cValues at a given stage for the same gene with different superscripts differ significantly (P<0.05).
Mentions: Generally, the improvement of DNA methylation reprogramming would effectively regulate gene expression [17]. Thus, the enhanced methylation reprogramming of Oct4 and Thy1 induced by 5-aza-dC or TSA should lead to their appropriate expression (Fig 3).

Bottom Line: In this study, we found that compared with in vitro fertilized counterparts, cloned embryos displayed the disrupted patterns of Oct4 demethylation and Thy1 remethylation.In conclusion, our results demonstrated that cloned embryos displayed incomplete gene-specific methylation reprogramming and disrupted expression patterns of pluripotency and tissue specific genes, and epigenetic modification agents improved gene-specific methylation reprogramming and expression pattern by regulating epigenetic modification related genes.This work would have important implications in improving cloning efficiency.

View Article: PubMed Central - PubMed

Affiliation: College of Animal Science and Technology, Qingdao Agricultural University, Qingdao, Shandong Province, China.

ABSTRACT
Incomplete DNA methylation reprogramming in cloned embryos leads to poor cloning efficiency. Epigenetic modification agents can improve genomic methylation reprogramming and the development of cloned embryos, however, the effect of epigenetic modification agents on gene-specific methylation reprogramming remains poorly studied. Here, we investigated DNA methylation reprogramming of pluripotency (Oct4) and tissue specific (Thy1) genes during early embryo development in pigs. In this study, we found that compared with in vitro fertilized counterparts, cloned embryos displayed the disrupted patterns of Oct4 demethylation and Thy1 remethylation. When 5-aza-2'-deoxycytidine (5-aza-dC) or trichostatin A (TSA) enhanced the development of cloned embryos, the transcripts of DNA methyltransferases (Dnmt1 and Dnmt3a), histone acetyltransferase 1 (Hat1) and histone deacetylase 1 (Hdac1) and the methylation and expression patterns of Oct4 and Thy1 became similar to those detected in in vitro fertilized counterparts. Further studies showed that Dnmt1 knockdown in cloned embryos enhanced the methylation reprogramming of Oct4 and Thy1 and promoted the activation of Oct4 and the silence of Thy1. In conclusion, our results demonstrated that cloned embryos displayed incomplete gene-specific methylation reprogramming and disrupted expression patterns of pluripotency and tissue specific genes, and epigenetic modification agents improved gene-specific methylation reprogramming and expression pattern by regulating epigenetic modification related genes. This work would have important implications in improving cloning efficiency.

No MeSH data available.


Related in: MedlinePlus