Global Methylation and Protamine Deficiency in Ram Spermatozoa Correlate with Sperm Production and Quality but Are Not Influenced by Melatonin or Season

SIMPLE SUMMARY: Though environmental factors can alter the epigenome of mammalian spermatozoa, it is currently unclear whether these epigenetic changes are linked to sperm production, quality and fertility. This study aimed to identify whether the hormone melatonin, responsible for upregulating ram reproductive function, is able to alter broad epigenetic markers in spermatozoa, namely sperm global methylation and protamine deficiency. It was also investigated whether these parameters corresponded to ram endocrinology, semen production and quality. Though no effects of season or melatonin were found, both sperm global methylation and protamine deficiency correlated with several semen production and quality parameters. These moderate associations with sperm production and quality support that sperm protamine deficiency and global methylation are broadly indicative of testicular function. ABSTRACT: This study assessed whether the seasonal effects of melatonin that upregulate ram reproductive function alter sperm global methylation or protamine deficiency and whether these parameters corresponded to ram endocrinology, semen production and quality. Ejaculates were assessed from rams that received melatonin implants (n = 9) or no implants (n = 9) during the non-breeding season. Ejaculates (n = 2/ram/week) were collected prior to implantation (week 0), 1, 6 and 12 weeks post implantation and during the following breeding season (week 30). Flow cytometry was used to assess the sperm global methylation and protamine deficiency in each ejaculate, which had known values for sperm concentration, motility, morphology, DNA fragmentation, seminal plasma levels of melatonin, anti-Mullerian hormone and inhibin A. Serum levels of testosterone and melatonin were also evaluated. Though there was no effect of melatonin or season, sperm protamine deficiency was negatively correlated with sperm production and seminal plasma levels of anti-Mullerian hormone and positively correlated with sperm DNA fragmentation and morphology. Global methylation of spermatozoa was positively correlated with sperm DNA fragmentation, morphology and serum testosterone and negatively correlated with sperm motility. These moderate associations with sperm production and quality suggest that sperm protamine deficiency and global methylation are indicative of ram testicular function.