Investigation on the mechanisms of human sperm DNA damage based on the proteomics analysis by SWATH-MS

Chun-Hui Zhu; Ye Wei; Fang Chen; Feng Li; Sheng-Min Zhang; Nai-Jun Dong; Tong-Min Xue; Kai-Feng Liu; Heng-Mi Cui; Jin-Chun Lu

doi:10.1186/s12014-022-09391-9

Investigation on the mechanisms of human sperm DNA damage based on the proteomics analysis by SWATH-MS

Clin Proteomics. 2023 Jan 6;20(1):2. doi: 10.1186/s12014-022-09391-9.

Authors

Chun-Hui Zhu^{1

2}, Ye Wei², Fang Chen¹, Feng Li¹, Sheng-Min Zhang¹, Nai-Jun Dong¹, Tong-Min Xue¹, Kai-Feng Liu¹, Heng-Mi Cui³, Jin-Chun Lu⁴

Affiliations

¹ Center for Reproductive Medicine, Northern Jiangsu People's Hospital Affiliated to Yangzhou University/Clinical Medical College, Yangzhou University, Yangzhou, 225001, Jiangsu, China.
² Institute of Epigenetics and Epigenomics, College of Animal Science and Technology, Yangzhou University, 48 Wenhui Road, Yangzhou, 225009, Jiangsu, China.
³ Institute of Epigenetics and Epigenomics, College of Animal Science and Technology, Yangzhou University, 48 Wenhui Road, Yangzhou, 225009, Jiangsu, China. hmcui@yzu.edu.cn.
⁴ Center for Reproductive Medicine, Zhongda Hospital, Southeast University, 3 Xinmofan Road, Nanjing, 210037, Jiangsu, China. 406646227@qq.com.

Abstract

Background: Spermatozoa have the task of delivering an intact paternal genome to the oocyte and supporting successful embryo development. The detection of sperm DNA fragmentation (SDF) has been emerging as a complementary test to conventional semen analysis for male infertility evaluation, but the mechanism leading to SDF and its impact on assisted reproduction remain unclear. Therefore, the study identified and analyzed the differentially expressed proteins of sperm with high and low SDF.

Methods: Semen samples from men attended the infertility clinic during June 2020 and August 2020 were analyzed, and sperm DNA fragmentation index (DFI) was detected by the sperm chromatin structure assay. Semen samples with low DFI (< 30%, control group) and high DFI (≥ 30%, experimental group) were optimized by density gradient centrifugation (DGC), and the differentially expressed proteins of obtained sperm were identified by the Sequential Window Acquisition of All Theoretical Mass Spectra Mass Spectrometry (SWATH-MS) and performed GO and KEGG analysis.

Results: A total of 2186 proteins were identified and 1591 proteins were quantified, of which 252 proteins were identified as differentially expressed proteins, including 124 upregulated and 128 downregulated. These differentially expressed proteins were involved in metabolic pathways, replication/recombination/repair, acrosomal vesicles, kinase regulators, fertilization, tyrosine metabolism, etc. Western blotting results showed that the expression levels of RAD23B and DFFA proteins and the levels of posttranslational ubiquitination and acetylation modifications in the experimental group were significantly higher than those in the control group, which was consistent with the results of proteomics analysis.

Conclusions: Proteomic markers of sperm with high DNA fragmentation can be identified by the SWATH-MS and bioinformatic analysis, and new protein markers and posttranslational modifications related to sperm DNA damage are expected to be intensively explored. Our findings may improve our understanding of the basic molecular mechanism of sperm DNA damage.

Keywords: Biomarker; Male infertility; Proteomics; Sperm DNA damage.

Abstract

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