Oocyte IVM or vitrification significantly impairs DNA methylation patterns in blastocysts as analysed by single-cell whole-genome methylation sequencing.
Oocyte IVM or vitrification significantly impairs DNA methylation patterns in blastocysts as analysed by single-cell whole-genome methylation sequencing.
复制标题
DOI：
10.1071/rd19234
复制
发表时间：
2020-04
期刊：
影响因子：
--
通讯作者：
Ya-Han Zhao;Jing-Jing Wang-Jing;Peipei Zhang;H. Hao;Yunwei Pang;Hao-Yu Wang;W. Du;Shanjiang Zhao
中科院分区：
文献类型：
--
作者：
Ya-Han Zhao;Jing-Jing Wang-Jing;Peipei Zhang;H. Hao;Yunwei Pang;Hao-Yu Wang;W. Du;Shanjiang Zhao
为探讨体外受精（IVF）囊胚质量差的机制，本研究采用单细胞全基因组甲基化测序技术，对牛体外受精（IVF）、新鲜（VIVF）和玻璃化冷冻（VIVF）囊胚的甲基化模式进行了分析。IVF组和VIVF组囊胚基因组甲基化水平均显著低于体内组（P<0.05）。总之，在IVF和体内组之间鉴定了1149个差异甲基化区域（DMR），在V_IVF和体内组之间鉴定了1578个DMR，在V_IVF和IVF组之间鉴定了151个DMR。印迹基因中胰岛素样生长因子2受体（IGF2R）和蛋白磷酸酶1调节亚基9A（PPP1R9A）甲基化水平在IVF组和V_IVF组均低于体内组，父系表达基因3（PEG3）甲基化水平在V_IVF组低于IVF组和体内组。IVF和体内以及V_IVF和IVF组之间具有DMR的基因主要富集在卵母细胞成熟途径中，而V_IVF和体内组之间的DMR富集在受精和玻璃化脆弱途径中。本研究的结果表明，关键DMR的甲基化差异可能导致体外和体内衍生胚胎之间的质量差异。
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 invivo, 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 invivo group (P<0.05). In all, 1149 differentially methylated regions (DMRs) were identified between the IVF and invivo groups, 1578 DMRs were identified between the V_IVF and invivo 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 invivo group, and the methylation level of paternally expressed 3 (PEG3) was lower in the V_IVF group than in the IVF and invivo groups. Genes with DMRs between the IVF and invivo and the V_IVF and IVF groups were primarily enriched in oocyte maturation pathways, whereas DMRs between the V_IVF and invivo 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 invitro- and invivo-derived embryos.