Resiliency of equid H19 imprint to somatic cell reprogramming by oocyte nuclear transfer and genetically induced pluripotency†.
Abstract: Cell reprogramming by somatic cell nuclear transfer and in induced pluripotent stem cells is associated with epigenetic modifications that are often incompatible with embryonic development and differentiation. For instance, aberrant DNA methylation patterns of the differentially methylated region and biallelic expression of H19-/IGF2-imprinted gene locus have been associated with abnormal growth of fetuses and placenta in several mammalian species. However, cloned horses are born with normal sizes and with no apparent placental anomalies, suggesting that H19/IGF2 imprinting may be epigenetically stable after reprogramming in this species. In light of this, we aimed at characterizing the equid H19 gene to determine whether imprinting is altered in somatic cell nuclear transfer (SCNT)-derived conceptuses and induced pluripotent stem cell (iPSC) lines using the mule hybrid model. A CpG-rich region containing five CTCF binding sites was identified upstream of the equine H19 gene and analyzed by bisulfite sequencing. Coupled with parent-specific and global H19 transcript analysis, we found that the imprinted H19 remains monoallelic and that on average the methylation levels of both parental differentially methylated regions in embryonic and extra-embryonic SCNT tissues and iPSC lines remained unaltered after reprogramming. Together, these results show that, compared to other species, equid somatic cells are more resilient to epigenetic alterations to the H19-imprinted locus during SCNT and iPSC reprogramming.
© The Author(s) 2019. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.
Publication Date: 2019-09-11 PubMed ID: 31504208DOI: 10.1093/biolre/ioz168Google Scholar: Lookup
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- Journal Article
Summary
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The research article represents an investigation into the resiliency of the H19 gene imprint within horse cells during the process of somatic cell nuclear transfer and generating induced pluripotent stem cell (iPSC) lines. The study reflects that equine cells display considerable resistance to epigenetic modifications in the H19 gene, contrary to what occurs in several other mammalian species.
Research Scope
- The aim of this research was to examine the equid H19 gene to ascertain if its imprinting is disrupted in somatic cell nuclear transfer-derived conceptuses and induced pluripotent stem cell lines.
- This study was conducted using a mule hybrid model.
Research Methodology
- The team identified a CpG-rich region encompassing five CTCF binding sites upstream of the equine H19 gene. This region was examined using bisulfite sequencing.
- Across parent-specific and global H19 transcript analysis, the researchers found that the imprinted H19 gene maintains its monoallelic status.
Research Findings
- Contrary to many other mammalian species, the average methylation levels of both parental differentially methylated regions within embryonic and extra-embryonic SCNT tissues as well as iPSC lines were unaltered by reprogramming.
- The equid somatic cells showed significant resilience to epigenetic changes to the H19-imprinted locus throughout the SCNT and iPSC reprogramming process.
Research Implications
- The findings suggest that cloned horses exhibit normal size, with no obvious abnormalities with their placenta, which possibly indicates that H19/IGF2 imprinting is likely to be epigenetically stable after reprogramming in equids.
- This comparative resilience to epigenetic modifications within the H19 gene in horse cells could have significant implications for the process of cell reprogramming and related applications within the field of veterinary medicine and associated fields.
Cite This Article
APA
Poirier M, Smith OE, Therrien J, Rigoglio NN, Miglino MA, Silva LA, Meirelles FV, Smith LC.
(2019).
Resiliency of equid H19 imprint to somatic cell reprogramming by oocyte nuclear transfer and genetically induced pluripotency†.
Biol Reprod, 102(1), 211-219.
https://doi.org/10.1093/biolre/ioz168 Publication
Researcher Affiliations
- Centre de Recherche en Reproduction et Fértilité, Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC, Canada.
- Centre de Recherche en Reproduction et Fértilité, Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC, Canada.
- Centre de Recherche en Reproduction et Fértilité, Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC, Canada.
- Centre de Recherche en Reproduction et Fértilité, Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC, Canada.
- Department of Surgery, School of Veterinary Medicine and Animal Science, University of Sao Paulo, São Paulo, SP, Brazil.
- Department of Surgery, School of Veterinary Medicine and Animal Science, University of Sao Paulo, São Paulo, SP, Brazil.
- Department of Veterinary, School of Animal and Food Sciences, University of São Paulo, Pirassununga, SP, Brazil.
- Department of Veterinary, School of Animal and Food Sciences, University of São Paulo, Pirassununga, SP, Brazil.
- Centre de Recherche en Reproduction et Fértilité, Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC, Canada.
MeSH Terms
- Animals
- Cellular Reprogramming / physiology
- Female
- Genomic Imprinting
- Horses
- Induced Pluripotent Stem Cells / metabolism
- Nuclear Transfer Techniques
- Oocytes / metabolism
- Ovary / metabolism
- RNA, Long Noncoding / genetics
- RNA, Long Noncoding / metabolism
Citations
This article has been cited 4 times.- Gastal GDA, Scarlet D, Melchert M, Ertl R, Aurich C. Epigenetic Changes in Equine Embryos after Short-Term Storage at Different Temperatures. Animals (Basel) 2021 May 6;11(5).
- Gambini A, Duque Rodríguez M, Rodríguez MB, Briski O, Flores Bragulat AP, Demergassi N, Losinno L, Salamone DF. Horse ooplasm supports in vitro preimplantation development of zebra ICSI and SCNT embryos without compromising YAP1 and SOX2 expression pattern. PLoS One 2020;15(9):e0238948.
- Zhang ZP, Zhang JT, Huang SC, He XY, Deng LX. Double sperm cloning (DSC) is a promising strategy in mammalian genetic engineering and stem cell research. Stem Cell Res Ther 2020 Sep 7;11(1):388.
- Luo M, Yi Y, Huang S, Dai S, Xie L, Liu K, Zhang S, Jiang T, Wang T, Yao B, Wang H, Xu D. Gestational dexamethasone exposure impacts hippocampal excitatory synaptic transmission and learning and memory function with transgenerational effects. Acta Pharm Sin B 2023 Sep;13(9):3708-3727.
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