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Updated: Sep 23, 2025
Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional Defects
Published on: April 21, 2022
Liquid chromatography-tandem mass spectrometry reveals an active response to DNA damage in human spermatozoa
Taylor Pini1, Mary Haywood1, Blair McCallie1
1Colorado Center for Reproductive Medicine, Lone Tree, Colorado.
Objective:
To investigate how endogenously elevated DNA fragmentation alters the human sperm proteome, and whether this fragmentation contributes to genomic deletions.
Design:
Research study.
Setting:
Commercial fertility clinic.
Patient(S):
Men with low (0%-4%, n = 7) or high (≥16%, n = 6) sperm DNA fragmentation, as assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling assay.
Intervention(S):
None.
Main Outcome Measure(S):
Global sperm proteome, single-nucleotide polymorphism genotyping array.
Result(S):
A total of 78 significantly differentially abundant proteins (30 decreased, 48 increased) were observed in control vs. high DNA damage samples. DNA damage resulted in robust proteomic responses, including markers of oxidative stress and apoptosis, DNA damage repair proteins, and transcription/translation and protein turnover machinery. Several key sperm functional proteins were significantly decreased in ejaculates with high DNA damage. We were unable to substantiate a link between increased DNA fragmentation and genomic deletions in human spermatozoa.
Conclusion(S):
Developing human spermatozoa initiate an active transcriptional response to endogenous DNA damage, which manifests as alterations in the sperm proteome.
Insights
High sperm DNA fragmentation alters the human sperm proteome, showing significant changes in protein levels. However, this study did not find a link between DNA fragmentation and genomic deletions in sperm.
Area of Science:
- Reproductive biology
- Molecular biology
- Genetics
Background:
- Sperm DNA fragmentation is a marker of sperm damage.
- Elevated DNA fragmentation can impact male fertility.
- The proteomic consequences of DNA fragmentation are not fully understood.
Purpose of the Study:
- To investigate how elevated sperm DNA fragmentation affects the human sperm proteome.
- To determine if sperm DNA fragmentation contributes to genomic deletions.
Main Methods:
- Proteomic analysis of sperm from men with low versus high DNA fragmentation.
- Terminal deoxynucleotidyl transferase dUTP nick end labeling assay for DNA fragmentation assessment.
- Single-nucleotide polymorphism genotyping array to assess genomic deletions.
Main Results:
- 78 differentially abundant proteins were identified in sperm with high DNA fragmentation.
- Proteomic alterations included markers of oxidative stress, apoptosis, and DNA repair.
- Key sperm functional proteins were decreased in samples with high DNA damage.
- No link was found between increased DNA fragmentation and genomic deletions.
Conclusions:
- Developing human spermatozoa exhibit a transcriptional response to endogenous DNA damage.
- This response leads to significant alterations in the sperm proteome.
- Sperm DNA fragmentation impacts sperm proteomic profiles without causing genomic deletions.
