ABSTRACT
The study of sperm epigenetics is challenging and limited largely to sperm population estimates rather than individual spermatozoa. While this type of approach is likely sufficient for most somatic cell lines, it is problematic in a tissue where a single cell is disproportionality influential. Furthermore, we know very little about the epigenetic variability between different sperm from the same ejaculate. Thus, it is essential that we better understand the heterogeneity of sperm epigenetic marks within an ejaculate. In this study, we have performed sperm genome-wide DNA methylation analyses on single ejaculates from 20 individuals. Sperm samples were subjected to gradient separation, following which the 90% layer ('high-quality sperm') and the 35% layer ('low-quality sperm') were isolated and analyzed separately using the Illumina 450K methylation array. We did not identify any single CpG that was differentially methylated between the two fractions. In contrast, we did identify 772 significant regional methylation alterations between the two layers. Coefficient of variance analysis also revealed that, in addition to having multiple sites that appear to be differentially methylated, the 35% layer sperm population, as a whole, displayed significantly higher variability in DNA methylation than did the 90% layer. In conclusion, while the two sperm populations analyzed here do not appear to be entirely distinct, those sperm that are generally considered to be of 'poor-quality' display some consistent regions of alteration and, more strikingly, demonstrate more heterogeneity than sperm considered to be of more normal quality.