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1.
Cell Reprogram ; 21(5): 260-269, 2019 10.
Artículo en Inglés | MEDLINE | ID: mdl-31596623

RESUMEN

DNA methylation is an essential epigenetic mark for embryo development and can be susceptible to environment factors such as in vitro conditions. The aim of this study was to verify the effect of in vitro culture until Day (D) 14 of the development on the embryo size and DNA methylation pattern of the insulin-like growth factor 2 (IGF2)-imprinted gene. To achieve this, we produced bovine embryos completely in vivo, completely in vitro, and in vitro until D7 and then in vivo up to D14. The embryos produced in in vitro were smaller than those in other two groups (p = 0.024); no differences in embryo size were observed between genders. The in vitro embryos showed a higher level of DNA methylation in the IGF2 as compared with that in the completely in vivo-produced (IVV) embryos (p = 0.009). Furthermore, totally in vitro-produced male embryos showed higher levels of DNA methylation as compared with those observed for the totally IVV male embryos (p = 0.034). No differences were observed among genders for IGF2 DNA methylation. These results showed that the window between D7 and D14 is critical for embryo development and alterations in the environmental conditions during this period can impair DNA methylation establishment of important developmental imprinted genes. This study brings unprecedented data for bovine embryos regarding the impact of the environmental conditions during the posthatching development.


Asunto(s)
Metilación de ADN , Técnicas de Cultivo de Embriones/métodos , Embrión de Mamíferos/citología , Desarrollo Embrionario/genética , Impresión Genómica , Factor II del Crecimiento Similar a la Insulina/genética , Trofoblastos/citología , Animales , Bovinos , Transferencia de Embrión , Embrión de Mamíferos/metabolismo , Femenino , Técnicas In Vitro , Masculino , Trofoblastos/metabolismo
2.
Placenta ; 70: 25-33, 2018 10.
Artículo en Inglés | MEDLINE | ID: mdl-30316323

RESUMEN

INTRODUCTION: Cloning via somatic cell nuclear transfer (SCNT) has been associated with a variety of pathologies, primarily in the placenta, and these alterations may be associated with aberrant epigenetic reprogramming of the donor cell genome. We tested the hypothesis that DNA methylation patterns are not appropriately established after nuclear transfer and that those altered patterns are associated with specific aberrant phenotypes. METHODS: We compared global and specific placental DNA methylation patterns between aberrant and healthy SCNT-produced calves. Foetal cotyledon samples of ten SCNT pregnancies were collected. Global DNA methylation and hydroxymethylation levels were measured using an ELISA-based assay and specific DNA methylation of satellite I, and α-satellite repeat elements were measured using bisulfite PCR. RESULTS: Our analysis revealed that the SCNT-produced calves, which showed aberrant phenotypes, exhibited a reduced methylation pattern of the satellite I region compared to that of healthy calves. In contrast, global methylation and hydroxymethylation analyses showed higher levels for both cytosine modifications in SCNT-produced female calves with aberrant phenotypes. The satellite I region showed most of the sequences to be hypermethylated in live cloned calves compared with those in deceased calves. DISCUSSION: Our results suggest that this satellite I region could be used as an epigenetic biomarker for predicting offspring viability. Studies evaluating DNA methylation patterns of this satellite region in the donor cell genome or embryo biopsies could shed light on how to improve the efficiency of SCNT cloning.


Asunto(s)
Metilación de ADN , Placenta/metabolismo , Placentación/fisiología , Animales , Bovinos , Clonación de Organismos , Epigénesis Genética , Femenino , Embarazo
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