Repeated assessment by high-throughput assay demonstrates that sperm DNA methylation levels are highly reproducible

Fertil Steril. 2011 Dec;96(6):1325-30. doi: 10.1016/j.fertnstert.2011.09.036. Epub 2011 Oct 26.

Abstract

Objective: To assess reliability of a high-throughput assay of sperm DNA methylation.

Design: Observational study comparing DNA methylation of sperm isolated from 3 divided and 12 longitudinally collected semen samples.

Setting: Academic medical center.

Patient(s): One man undergoing screening semen analysis during evaluation of an infertile couple and 2 healthy fertile male volunteers.

Intervention(s): Spermatozoa were separated from seminal plasma and somatic cells using gradient separation. DNA was extracted from spermatozoa, and DNA methylation was assessed at 1,505 DNA sequence-specific sites.

Main outcome measure(s): Repeatability of sperm DNA methylation measures, estimated by correlation coefficients.

Result(s): DNA methylation levels were highly correlated within matched sets of divided samples (all r ≥ 0.97) and longitudinal samples (average r = 0.97).

Conclusion(s): The described methodology reliably assessed methylation of sperm DNA at large numbers of sites. Methylation profiles were consistent over time. High-throughput assessment of sperm DNA methylation is a promising tool for studying the role of epigenetic state in male fertility.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Methylation* / physiology
  • High-Throughput Nucleotide Sequencing / methods
  • High-Throughput Nucleotide Sequencing / standards
  • High-Throughput Nucleotide Sequencing / statistics & numerical data*
  • Humans
  • Infertility, Male / genetics
  • Infertility, Male / metabolism
  • Longitudinal Studies
  • Male
  • Reproducibility of Results
  • Semen Analysis
  • Sensitivity and Specificity
  • Spermatozoa / cytology
  • Spermatozoa / metabolism*
  • Time Factors
  • Validation Studies as Topic