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Single Oocyte Bisulfite Mutagenesis
Published on: June 27, 2012
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Minor DNA methylation changes are observed in spermatozoa prepared using different protocols
Martin Stimpfel1, Eda Vrtacnik-Bokal1
1Department of Human Reproduction, Division of Gynaecology, University Medical Center Ljubljana, Ljubljana, Slovenia.
Andrology
|May 30, 2020
Summary
Sperm preparation methods impact DNA methylation profiles, with distinct differentially methylated regions observed between conventional density gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) techniques. Further research is needed to confirm these findings on sperm DNA methylation.
Area of Science:
- Reproductive Biology
- Epigenetics
- Genomics
Background:
- DNA methylation patterns are heritable and influenced by environmental factors.
- Assisted reproductive technologies may alter these epigenetic patterns.
Purpose of the Study:
- To compare the effects of two sperm preparation methods on sperm DNA methylation.
- Evaluate conventional density gradient centrifugation (DGC) versus DGC with magnetic-activated cell sorting (MACS).
Main Methods:
- Semen samples from IVF patients were divided and processed using DGC or DGC+MACS.
- Reduced representation bisulfite sequencing (RRBS) analyzed DNA methylation profiles.
- Sperm morphology and DNA fragmentation were also assessed.
Main Results:
- Genome-wide methylation levels were similar between DGC and MACS groups (0.53-0.56).
- 99 differentially methylated regions (DMRs) and 800 differentially methylated positions (DMPs) were identified.
- Three imprinting-associated genes (IGF2, PRDM16, CLF4/BRUNOL4) were identified among DMRs/DMPs.
Conclusions:
- Distinct methylation changes exist between DGC and MACS sperm preparation methods, despite similar overall methylation levels.
- Further studies with more samples and varied methods are recommended to validate findings on sperm DNA methylation.
