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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Oocyte IVM or vitrification significantly impairs DNA methylation patterns in blastocysts as analysed by single-cell whole-genome methylation sequencing

Ya-Han Zhao A * , Jing-Jing Wang A * , Pei-Pei Zhang A * , Hai-Sheng Hao A , Yun-Wei Pang A , Hao-Yu Wang A , Wei-Hua Du A , Shan-Jiang Zhao A , Wei-Min Ruan B , Hui-Ying Zou A , Tong Hao A , Hua-Bin Zhu A and Xue-Ming Zhao https://orcid.org/0000-0001-6110-2690 A C
+ Author Affiliations
- Author Affiliations

A Embryo Biotechnology and Reproduction Laboratory and the Centre of Domestic Animal Reproduction and Breeding, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan Western Road, Haidian District, Beijing 100193, PR China.

B International Joint Center for Biomedical Innovation, School of Life Sciences, Henan University, Ming Lun Street, Kaifeng, Henan, 475004, PR China.

C Corresponding author. Email: zhaoxueming@caas.cn

Reproduction, Fertility and Development 32(7) 676-689 https://doi.org/10.1071/RD19234
Submitted: 26 June 2019  Accepted: 14 November 2019   Published: 11 March 2020

Abstract

To explore the mechanisms leading to the poor quality of IVF blastocysts, the single-cell whole-genome methylation sequencing technique was used in this study to analyse the methylation patterns of bovine blastocysts derived from in vivo, fresh (IVF) or vitrified (V_IVF) oocytes. Genome methylation levels of blastocysts in the IVF and V_IVF groups were significantly lower than those of the in vivo group (P < 0.05). In all, 1149 differentially methylated regions (DMRs) were identified between the IVF and in vivo groups, 1578 DMRs were identified between the V_IVF and in vivo groups and 151 DMRs were identified between the V_IVF and IVF groups. For imprinted genes, methylation levels of insulin-like growth factor 2 receptor (IGF2R) and protein phosphatase 1 regulatory subunit 9A (PPP1R9A) were lower in the IVF and V_IVF groups than in the in vivo group, and the methylation level of paternally expressed 3 (PEG3) was lower in the V_IVF group than in the IVF and in vivo groups. Genes with DMRs between the IVF and in vivo and the V_IVF and IVF groups were primarily enriched in oocyte maturation pathways, whereas DMRs between the V_IVF and in vivo groups were enriched in fertilisation and vitrification-vulnerable pathways. The results of this study indicate that differences in the methylation of critical DMRs may contribute to the differences in quality between in vitro- and in vivo-derived embryos.

Graphical Abstract Image

Additional keywords: bovine, development, embryo, mechanism.


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