Transcriptome analysis of human spermatozoa with different DNA fragmentation index using RNA sequencing

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology Pub Date : 2024-11-04 DOI:10.1016/j.repbio.2024.100964

Kailin Yang , Xue Sun , Qiyuan Zheng , Chen Pan , Siyuan Wang , Qingfang Lu , Changlong Xu , Yangqing Lu

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引用次数: 0

Abstract

The study is aimed to screen the differential expressed genes (DEGs) related to sperm DNA fragmentation in men and provide reference basis of the sperm selection in assisted reproduction based on DNA fragmentation. We evaluated 60 semen samples from patients with high, medium or low sperm DNA fragmentation index (DFI). Using multicolor flow cytometry, we measured the content of reactive oxygen species (ROS), phosphatidylserine (PS) externalization and mitochondrial membrane potential (MMP) in these sperm samples. The results revealed that the more ROS content and PS externalization were detected in the sperm with higher DFI, but there was lower MMP level in the high DFI sperm. Next, we conducted RNA sequencing (RNA-seq) on 3 groups of sperm samples with high, medium and low DFI. Then, Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were performed on DEGs. Furthermore, we utilized qRT-PCR to validated the significantly DEGs from the RNA-seq assay. The transcriptome results showed a total of 5334 DEGs were found in the sperm sample with high, medium and low DFI. According to GO and KEGG analysis 421 down-regulated genes in the high DFI group were related to oxidative stress and spermatogenesis. Thirteen novel genes were also identified that most likely were involved in sperm DNA fragmentation, which were further validated by qRT-PCR. In conclusion, our study suggested that the sperms with highly fragmented DNA were accompanied by down-regulation of a series of genes related to antioxidant and spermatogenesis.

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利用 RNA 测序分析不同 DNA 破碎指数的人类精子转录组

本研究旨在筛选与男性精子DNA碎片相关的差异表达基因（DEGs），为辅助生殖中根据DNA碎片选择精子提供参考依据。我们评估了60份精子DNA碎片指数（DFI）为高、中、低的患者精液样本。我们使用多色流式细胞术测量了这些精子样本中活性氧（ROS）、磷脂酰丝氨酸（PS）外化和线粒体膜电位（MMP）的含量。结果显示，DFI较高的精子中ROS含量和磷脂酰丝氨酸（PS）外化程度较高，但DFI较高的精子中线粒体膜电位（MMP）水平较低。接着，我们对高、中、低DFI的三组精子样本进行了RNA测序（RNA-seq）。然后，我们对 DEGs 进行了基因本体（GO）富集分析和京都基因组百科全书（KEGG）通路分析。此外，我们还利用 qRT-PCR 验证了 RNA-seq 分析中显著的 DEGs。转录组结果显示，高、中、低DFI精子样本中共发现了5334个DEGs。根据 GO 和 KEGG 分析，高 DFI 组中有 421 个下调基因与氧化应激和精子发生有关。研究还发现了 13 个很可能与精子 DNA 断裂有关的新基因，并通过 qRT-PCR 对其进行了进一步验证。总之，我们的研究表明，DNA高度破碎的精子伴随着一系列与抗氧化和精子发生有关的基因的下调。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Reproductive biology 生物-生殖生物学

CiteScore

3.90

自引率

0.00%

发文量

审稿时长

29 days

期刊介绍： An official journal of the Society for Biology of Reproduction and the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn, Poland. Reproductive Biology is an international, peer-reviewed journal covering all aspects of reproduction in vertebrates. The journal invites original research papers, short communications, review articles and commentaries dealing with reproductive physiology, endocrinology, immunology, molecular and cellular biology, receptor studies, animal breeding as well as andrology, embryology, infertility, assisted reproduction and contraception. Papers from both basic and clinical research will be considered.