Optimizing 5-aza-2'-deoxycytidine treatment to enhance the development of porcine cloned embryos by inhibiting apoptosis and improving DNA methylation reprogramming.
Animals
Apoptosis
/ drug effects
Azacitidine
/ pharmacology
Blastocyst
/ metabolism
Cellular Reprogramming
/ drug effects
Cloning, Organism
/ methods
DNA Methylation
/ drug effects
Decitabine
/ pharmacology
Embryonic Development
/ drug effects
Gene Expression Regulation, Developmental
/ drug effects
Insulin-Like Growth Factor II
/ genetics
Nuclear Transfer Techniques
/ veterinary
Swine
/ embryology
5-aza-dC
Apoptosis
Cloned embryos
DNA methylation
Pig
Journal
Research in veterinary science
ISSN: 1532-2661
Titre abrégé: Res Vet Sci
Pays: England
ID NLM: 0401300
Informations de publication
Date de publication:
Oct 2020
Oct 2020
Historique:
received:
07
12
2019
revised:
18
06
2020
accepted:
18
06
2020
pubmed:
4
7
2020
medline:
1
1
2021
entrez:
4
7
2020
Statut:
ppublish
Résumé
Apoptosis and incomplete DNA methylation reprogramming in cloned embryos reduce cloning efficiency. 5-aza-2'-deoxycytidine (5-aza-dC) is proven to regulate apoptosis and DNA methylation reprogramming, however, the treatment method and potential role of 5-aza-dC during cloned embryo development are still not well studied. This study displayed that treating donor cells with 5-aza-dC (AN group) significantly reduced the blastocyst rate, while treating cloned embryos (NA group) or both donor cells and cloned embryos (ANA group) significantly promoted the blastocyst formation, and the ANA group was the best treatment of 5-aza-dC to enhance the development of cloned embryos. Then, compared with the NT group, the ANA group showed the significantly enhanced nuclear remodeling. The apoptotic cell numbers and rates of blastocysts were significantly reduced, and the expression levels of significantly upregulated anti-apoptosis gene Bcl2l1 and downregulated pro-apoptosis genes Bax, P53 and Caspase3 were observed in the ANA group. Further study demonstrated that the transcription levels of DNA methylation reprogramming genes Dnmt1, Dnmt3a, Tet1 and Tet3 were significantly upregulated, and, significant genomic DNA remethylation, DNA demethylation of pluripotency gene Oct4, and DNA remethylation of tissue specific gene Thy1 were observed at the blastocyst stage in the ANA group. Embryo development related genes including Igf2, H19, Oct4, Nanog, Sox2, Eif1a, Cdx2 and ATP1b1 were significantly upregulated, and Thy1 and Col5a2 were remarkably silenced at the 4-cell and blastocyst stages in the ANA group. In conclusion, the best 5-aza-dC treatment enhanced the development of cloned embryos by inhibiting apoptosis and improving DNA methylation reprogramming.
Identifiants
pubmed: 32619801
pii: S0034-5288(20)30911-5
doi: 10.1016/j.rvsc.2020.06.020
pii:
doi:
Substances chimiques
Insulin-Like Growth Factor II
67763-97-7
Decitabine
776B62CQ27
Azacitidine
M801H13NRU
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
229-236Informations de copyright
Copyright © 2020 Elsevier Ltd. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Competing Interest The authors declare that no conflicting financial interests exist.