Zhu Haibo, Sun Hao, Yu Dawei, Li Tianda, Hai Tang, Liu Chao, Zhang Ying, Chen Yurong, Dai Xiangpeng, Li Ziyi, Li Wei, Liu Ruizhi, Feng Guihai, Zhou Qi
Center of Reproductive Medicine, Center of Prenatal Diagnosis, First Hospital, Jilin University, Changchun, China.
State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Front Cell Dev Biol. 2021 Mar 16;9:632183. doi: 10.3389/fcell.2021.632183. eCollection 2021.
Low birth efficiency and developmental abnormalities in embryos derived using round spermatid injection (ROSI) limit the clinical application of this method. Further, the underlying molecular mechanisms remain elusive and warrant further in-depth study. In this study, the embryonic day (E) 11.5 mouse fetuses and corresponding placentas derived upon using ROSI, intracytoplasmic sperm injection (ICSI), and natural fertilized (control) embryos were collected. Transcriptome and DNA methylation profiles were analyzed and compared using RNA-sequencing (RNA-seq) and whole-genome bisulfite sequencing, respectively. RNA-seq results revealed similar gene expression profiles in the ROSI, ICSI, and control fetuses and placentas. Compared with the other two groups, seven differentially expressed genes (DEGs) were identified in ROSI fetuses, and ten DEGs were identified in the corresponding placentas. However, no differences in CpG methylation were observed in fetuses and placentas from the three groups. Imprinting control region methylation and imprinted gene expression were the same between the three fetus and placenta groups. Although 49 repetitive DNA sequences (RS) were abnormally activated in ROSI fetuses, RS DNA methylation did not differ between the three groups. Interestingly, abnormal hypermethylation in promoter regions and low expression of and were correlated with a crown-rump length less than 6 mm in one ROSI fetus. Our study demonstrates that the transcriptome and DNA methylation in ROSI-derived E11.5 mouse fetuses and placentas were comparable with those in the other two groups. However, some abnormally expressed genes in the ROSI fetus and placenta warrant further investigation to elucidate their effect on the development of ROSI-derived embryos.
利用圆形精子细胞注射（ROSI）获得的胚胎出生效率低且存在发育异常，这限制了该方法的临床应用。此外，其潜在的分子机制仍不清楚，需要进一步深入研究。在本研究中，收集了使用ROSI、卵胞浆内单精子注射（ICSI）和自然受精（对照）胚胎产生的胚胎第11.5天（E11.5）的小鼠胎儿及相应胎盘。分别使用RNA测序（RNA-seq）和全基因组亚硫酸氢盐测序分析并比较了转录组和DNA甲基化谱。RNA-seq结果显示，ROSI、ICSI和对照胎儿及胎盘的基因表达谱相似。与其他两组相比，在ROSI胎儿中鉴定出7个差异表达基因（DEG），在相应胎盘中鉴定出10个DEG。然而，三组胎儿和胎盘的CpG甲基化未观察到差异。三组胎儿和胎盘的印记控制区甲基化和印记基因表达相同。虽然在ROSI胎儿中有49个重复DNA序列（RS）异常激活，但三组之间RS DNA甲基化没有差异。有趣的是，在一个ROSI胎儿中，启动子区域的异常高甲基化以及[具体基因]和[具体基因]的低表达与头臀长度小于6 mm相关。我们的研究表明，ROSI来源的E11.5小鼠胎儿和胎盘的转录组和DNA甲基化与其他两组相当。然而，ROSI胎儿和胎盘中一些异常表达的基因值得进一步研究，以阐明它们对ROSI来源胚胎发育的影响。
