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Updated: Jun 8, 2025
08:48
Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional Defects
Published on: April 21, 2022
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Transcriptome analysis of human spermatozoa with different DNA fragmentation index using RNA sequencing
Kailin Yang1, Xue Sun2, Qiyuan Zheng1
1Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, College of Animal Science and Technology, Guangxi University, Nanning, Guangxi, China.
Reproductive Biology
|November 5, 2024
Summary
Sperm DNA fragmentation in men is linked to increased reactive oxygen species (ROS) and decreased mitochondrial potential. Gene expression analysis revealed key genes related to oxidative stress and spermatogenesis are downregulated in sperm with high DNA fragmentation.
Area of Science:
- Reproductive Biology
- Genomics
- Oxidative Stress Research
Background:
- Sperm DNA fragmentation (SDF) negatively impacts male fertility and assisted reproduction outcomes.
- Understanding the molecular mechanisms underlying SDF is crucial for improving sperm selection and treatment strategies.
Purpose of the Study:
- To identify differentially expressed genes (DEGs) associated with sperm DNA fragmentation (SDF) in men.
- To provide a molecular basis for sperm selection in assisted reproduction based on DNA fragmentation levels.
Main Methods:
- Evaluation of 60 semen samples categorized by high, medium, and low sperm DNA fragmentation index (DFI).
- Assessment of reactive oxygen species (ROS) content, phosphatidylserine (PS) externalization, and mitochondrial membrane potential (MMP) using multicolor flow cytometry.
- RNA sequencing (RNA-seq) followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of DEGs.
- Validation of key DEGs using quantitative reverse transcription PCR (qRT-PCR).
Main Results:
- Sperm with higher DFI exhibited increased ROS content and PS externalization, alongside decreased MMP.
- RNA-seq identified 5334 DEGs across high, medium, and low DFI groups.
- GO and KEGG analyses revealed 421 downregulated genes in the high DFI group associated with oxidative stress and spermatogenesis.
- Thirteen novel genes implicated in SDF were identified and validated via qRT-PCR.
Conclusions:
- Highly fragmented sperm DNA is associated with the downregulation of genes critical for antioxidant defense and spermatogenesis.
- The identified DEGs offer potential biomarkers for assessing sperm quality and guiding assisted reproduction techniques.
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