Loss of methylation of H19-imprinted gene derived from assisted reproductive technologies can be mitigated by cleavage-stage embryo transfer in mice
Loss of methylation of H19-imprinted gene derived from assisted reproductive technologies can be mitigated by cleavage-stage embryo transfer in mice
复制标题
通过小鼠卵裂期胚胎移植可以减轻源自辅助生殖技术的 H19 印记基因甲基化的丧失
DOI：
10.1007/s10815-019-01575-x
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发表时间：
2019-11-01
影响因子：
3.1
通讯作者：
Li, Bo
中科院分区：
文献类型：
--
作者：
Chen, Shuqiang;Zhang, Meizi;Li, Bo
目的：对啮齿动物的研究表明，辅助生殖技术（ART）与囊胚期胚胎基因组印记的干扰有关。然而，ART对胚胎基因组印记影响的脆弱发育窗口仍未确定。本研究的目的是建立特定的胚胎发育阶段，在该阶段，H19印迹控制区（ICR）的甲基化丢失是由ART发生。此外，我们探讨了协议，以防止可能的负面影响ART胚胎H19 imprinting.Methods：小鼠胚胎产生四种不同的实验条件下，分为四组：对照，体外培养（IVC），体外受精（IVF），卵胞浆内单精子注射（ICSI）。结果：IVC、IVF和ICSI对小鼠囊胚H19 ICR基因甲基化水平的影响程度相似。具体而言，我们观察到在小鼠8-细胞和桑椹胚阶段之间H19 ICR的甲基化的显著损失。此外，我们发现，在8-细胞阶段的子宫中的小鼠胚胎移植产生的ART诱导的囊胚中的甲基化模式的发生更接近在体内的thes.Conclusions：我们的研究结果表明，由ART引起的H19 ICR的甲基化损失发生在8-细胞和桑椹胚阶段之间，和卵裂胚胎转移到子宫减轻了由小鼠ART衍生的H19的甲基化损失。
Purpose:Studies on rodents have shown that assisted reproductive technologies (ARTs) are associated with perturbation of genomic imprinting in blastocyst-stage embryos. However, the vulnerable developmental window for ART influence on the genomic imprinting of embryos is still undetermined. The purpose of this study was to establish the specific embryonic development stage at which the loss of methylation of H19 imprinting control regions (ICRs) was caused by ART occurrence. Additionally, we explored protocols to safeguard against possible negative impacts of ART on embryo H19 imprinting.Methods:Mouse embryos were generated under four different experimental conditions, divided into four groups: control, in vitro culture (IVC), in vitro fertilization (IVF), and intracytoplasmic sperm injection (ICSI). The methylation levels of H19 ICR of the grouped or individual embryos were analyzed by bisulfite-sequencing PCR.Results:Our data showed that the loss of methylation of H19 ICR in mouse blastocysts was inflicted to a similar extent by IVC, IVF, and ICSI. Specifically, we observed a significant loss of methylation of H19 ICR between the mouse 8-cell and morula stages. In addition, we revealed that the transfer of mouse embryos generated by ARTs in the uterus at the 8-cell stage induced the occurrence of methylation patterns in the blastocysts closer to the in vivo ones.Conclusions:Our findings indicate that the loss of methylation of H19 ICR caused by ARTs occurs between the 8-cell and the morula stages, and the transfer of cleavage embryos to the uterus mitigates the loss methylation of H19 derived by mice ARTs.