Methylation changes in mature sperm deoxyribonucleic acid from oligozoospermic men: assessment of genetic variants and assisted reproductive technology.

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Methylation changes in mature sperm deoxyribonucleic acid from oligozoospermic men: assessment of genetic variants and assisted reproductive technology outcome Debbie Montjean, Ph.D., Célia Ravel, M.D., Ph.D., Moncef Benkhalifa, Ph.D., Paul Cohen-Bacrie, D.Pharm., Isabelle Berthaut, Ph.D., Anu Bashamboo, Ph.D., Kenneth McElreavey, Ph.D. Fertility and Sterility Volume 100, Issue 5, Pages 1241-1247.e2 (November 2013) DOI: 10.1016/j.fertnstert.2013.06.047 Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Summary of methylation errors found the H19- and PEG1/MEST-DMR in sperm DNA of infertile men described in Table 1. The DNA of “normal” sperm cells is expected to be fully methylated at the H19-DMR. Top: methylation profile in the H19-DMR in oligozoospermic patients. The numbers at top, above the circles (1–18), represent individual CpGs in the PCR amplicon. Each black circle represents a methylated state of that particular CpG in all clones analyzed (n > 50), whereas white circles represent hypomethylated CpGs. The right column shows the average level of methylation at each hypomethylated CpG. Example: the H19-DMR in patient 1’s sperm DNA is hypomethylated on three sites, namely CpG4, CpG6, and CpG13. The methylation level on those CpGs is 60%, 27%, and 86%, respectively. Methylation of the seventh CpG grey circle is variable in the general population because there is a C/T polymorphism at this site, and hence it was excluded from the study. Bottom: methylation profile in the PEG1/MEST-DMR in oligozoospermic men. The DNA of “normal” sperm cells is expected to be fully unmethylated at the PEG1/MEST-DMR. The numbers at top, above the circles (1–15), represent individual CpGs in the PCR amplicon. Each black circle represents a methylated state of that particular CpG in all clones analyzed (n > 50), whereas white circles represent hypomethylated CpGs. The right column shows the average level of methylation at each hypermethylated CpG. Fertility and Sterility 2013 100, 1241-1247.e2DOI: (10.1016/j.fertnstert.2013.06.047) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Structural characteristics of the human H19-DMR and MEST/PEG1-DMR loci analyzed in this study. H19-DMR and MEST/PEG1-DMR methylation analysis involved 18 and 15 CpGs, respectively. Black circles: methylated CpGs. White circles: unmethylated CpGs. The transcription start site is indicated by an arrow. Differentially methylated regions (DMRs) are represented by grey boxes. Fertility and Sterility 2013 100, 1241-1247.e2DOI: (10.1016/j.fertnstert.2013.06.047) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions