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Phosphopeptide Analysis of Rodent Epididymal Spermatozoa
Published on: December 30, 2014
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Proteomic Landscape of Human Spermatozoa: Optimized Extraction Method and Application.
Mengqi Luo1, Tao Su1, Shisheng Wang1
1Laboratory of Clinical Proteomics and Metabolomics, Institutes for Systems Genetics, Frontiers Science Center for Disease-Related Molecular Network, West China Hospital, Sichuan University, Chengdu 610017, China.
Cells
|December 23, 2022
Summary
Optimizing human sperm protein extraction is crucial. A new method using urea buffer and ultrasonication achieved the deepest proteome coverage, revealing antibiotics may impact sperm quality and morphology.
Area of Science:
- Proteomics
- Spermatozoa Biology
- Analytical Chemistry
Background:
- Understanding human spermatozoa proteomics is vital for reproductive health.
- Existing sample preparation methods limit in-depth protein analysis of sperm cells.
- The impact of antibiotics on sperm proteome and quality remains unclear.
Purpose of the Study:
- To compare and optimize protein extraction methods for human sperm.
- To investigate the effects of antibiotics (amoxicillin, clarithromycin) on sperm proteome.
- To assess changes in sperm quality and morphology during antibiotic therapy.
Main Methods:
- Systematic comparison of six protein extraction methods combining lysis buffers and physical lysis.
- Identification of the optimal method: urea buffer with ultrasonication (UA-ultrasonication).
- Longitudinal sperm proteome analysis using data-independent acquisition tandem mass spectrometry (DIA-MS/MS) during antibiotic treatment for H. pylori infection.
Main Results:
- UA-ultrasonication yielded the highest protein extraction rate and deepest human sperm proteome coverage (5685 protein groups).
- Antibiotic therapy correlated with increased sperm concentration and malformed sperm rates.
- Significant disruption of sperm proteome expression was observed during antibiotic treatment.
Conclusions:
- The UA-ultrasonication method offers an optimized approach for in-depth human sperm proteome characterization.
- Antibiotics may negatively affect sperm quality and proteome integrity.
- This optimized method has potential for expanded clinical applications in reproductive medicine.
