Journal of Assisted Reproduction and Genetics ( IF 3.1 ) Pub Date : 2020-06-23 , DOI: 10.1007/s10815-020-01839-x Wanhong He 1 , Υuhua Sun 2 , Sufen Zhang 1 , Xing Feng 2 , Minjie Xu 1 , Jianfeng Dai 3 , Xiaohua Ni 1 , Xin Wang 2 , Qihan Wu 1
Purpose
Changes in DNA methylation modifications have been associated with male infertility. With the development of assisted reproductive technologies (ARTs), abnormal DNA methylation in sperm, especially in imprinted genes, may impact the health of offspring and requires an in-depth study.
Methods
In this study, we collected abnormal human semen samples, including asthenospermic, oligospermic, oligoasthenospermic and deformed sperm, and investigated the methylation of imprinted genes by reduced representation bisulfite sequencing (RRBS) and bisulfite amplicon sequencing on the Illumina platform.
Results
The differentially methylated regions (DMRs) of imprinted genes, including H19, GNAS, MEG8 and SNRPN, were different in the abnormal semen groups. MEG8 DMR methylation in the asthenospermic group was significantly increased. Furthermore, higher methylation levels of MEG8, GNAS and SNRPN DMR in the oligospermic and oligoasthenospermic groups and a decrease in the H19 DMR methylation level in the oligospermic group were observed. However, the methylation levels of these regions varied greatly among the different semen samples and among individual sperm within the same semen sample. The SNP rs2525883 genotype in the H19 DMR affected DNA methylation. Moreover, DNA methylation levels differed in the abnormal semen groups in the non-imprinted genomic regions, including repetitive sequence DNA transposons and long/short interspersed nuclear elements (LINEs and SINEs).
Conclusion
Our study established that imprinted gene DMRs, such as H19, GNAS, SNRPN and MEG8, were differentially methylated in the abnormal semen groups with obvious inter- and intra-sample heterogeneities. These results suggest that special attention needs to be paid to possible epigenetic risks during reproduction.
中文翻译:
通过下一代亚硫酸氢盐测序分析异常精液样本中印记基因的 DNA 甲基化模式。
目的
DNA 甲基化修饰的变化与男性不育有关。随着辅助生殖技术(ARTs)的发展,精子中的异常DNA甲基化,特别是印迹基因,可能会影响后代的健康,需要深入研究。
方法
在这项研究中,我们收集了异常人类精液样本,包括弱精子、少精子、少弱精子和畸形精子,并通过减少代表性亚硫酸氢盐测序(RRBS)和亚硫酸氢盐扩增子测序在 Illumina 平台上研究了印记基因的甲基化。
结果
的差异甲基化区域印记基因,包括(DMRS)H19,GNAS,MEG8和SNRPN,分别在精液异常基团不同。弱精子症组的MEG8 DMR 甲基化显着增加。此外,较高的甲基化水平MEG8,GNAS和SNRPN在少精子和oligoasthenospermic基DMR和在降低H19观察到少精组中的 DMR 甲基化水平。然而,这些区域的甲基化水平在不同的精液样本之间以及在同一精液样本中的单个精子之间差异很大。H19 DMR 中的 SNP rs2525883 基因型影响 DNA 甲基化。此外,非印迹基因组区域中异常精液组的 DNA 甲基化水平不同,包括重复序列 DNA 转座子和长/短散布核元件(LINEs和SINEs)。
结论
我们的研究证实,印记基因的DMR,诸如H19,GNAS,SNRPN和MEG8,被差异甲基化在精液异常组具有明显之间和内部样品的非均质性。这些结果表明需要特别注意生殖过程中可能存在的表观遗传风险。