JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2017, Vol. 55 ›› Issue (1): 54-59.doi: 10.6040/j.issn.1671-7554.0.2016.1065

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DNA methylation and expression of SNRPN and GRB10 imprinted genes in human semen freezing-thawing process

ZHOU Xue1, WANG Yanrong1, TIAN Long1, MA Lianghong2, YAN Bei2, TIAN Jia2, ZHANG Fan2, ZHOU Yue2, WANG Hongyan1,2   

  1. 1. Key Laboratory of Fertility Maintenance, Ministry of Education, Ningxia Medical University &
    Key Laboratory of Reproduction and Genetics;
    2. General Hospital of Ningxia Medical University, Ningxia Human Sperm Bank, Ningxia 750004, Yinchuan, China
  • Received:2016-08-29 Online:2017-01-10 Published:2017-01-10

Abstract: Objective To assess the effect of human semen frozen-thawing process on sperm DNA methylation status and mRNA expressions of SNRPN and GRB10 imprinted genes. Methods The semen samples were collected from 20 healthy men, and each sample was divided into two equalaliquots: fresh group(n=20)and cryopreserved group(n=20). Routine semen parameters was tested by computer-aided sperm analysis(CASA). Each semen sample was purificated by Percoll density gradient centrifugation. The DNA methylation status of SNRPN promoter region neighbouring fragment and GRB10 promoter region fragment were determined by Sequenom DNA sequencing. The mRNA expressions of SNRPN and GRB10 were evaluated by qRT-PCR. Results Compared with fresh group,the semen parameters including sperm concentration, percentage of sperm motility and forward-moving sperms all decreased in cryopreserved 山 东 大 学 学 报 (医 学 版)55卷1期 -周雪,等.冷冻复苏过程对人精子印记基因SNRPN和GRB10DNA甲基化及表达的影响 \=-group(P<0.05), and percentage of unforward-moving sperm increased significantly(P<0.05). Compared with fresh group, the mRNA expression of SNRPN decreased significantly in cryopreserved group(P<0.05). DNA methylation status on the forth methylation site(-902CpG point)of GRB10 promoter region fragment remarkably decreased(P<0.05)and the mRNA expression of GRB10 increased significantly in cryopreserved group(P<0.05). Conclusion The freezing-thawing process affects DNA methylation status and mRNA expressions of human sperm SNRPN and GRB10 imprinted genes. It suggests that the semen freezing technique routinely used in the human sperm bank may alter the epigenetic information of the gametes and consequently regulate the expression of related genes.

Key words: DNA methylation, Growth factor receptor-binding protein 10, Small nuclear ribonucleoprotein polypeptide N gene, Semen freezing technique

CLC Number: 

  • R321.1
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