Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 22, Issue 5, Pages -Publisher
MDPI
DOI: 10.3390/ijms22052679
Keywords
spermatozoa; genotyping by sequencing (GBS); methylated DNA immunoprecipitation (MeDIP); artificial insemination; fertility; prolificacy; pig
Funding
- Research Council FORMAS, Stockholm [201700946, 2018-01074, 2019-00288]
- MINECO (Spain)
- FEDER funds (EU) [AGL2015-69738-R]
- Seneca Foundation Murcia, Spain [19892/GERM-15]
- FAPESP [2016/20440-3, 2018/13600-0]
- Formas [2018-01074] Funding Source: Formas
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A combined GBS and MeDIP protocol was used to identify genetic and epigenetic differences in the genome of spermatozoa from porcine animal models. The study found that breeding boars with different fertility levels exhibited significant differences in their genetic and epigenetic profiles, with potential applications in sire selection and diagnosis of reproductive system diseases in males. This suggests that methylome analyses can be used to discern fertility levels in breeding males.
A combined Genotyping By Sequencing (GBS) and methylated DNA immunoprecipitation (MeDIP) protocol was used to identify-in parallel-genetic variation (Genomic-Wide Association Studies (GWAS) and epigenetic differences of Differentially Methylated Regions (DMR) in the genome of spermatozoa from the porcine animal model. Breeding boars with good semen quality (n = 11) and specific and well-documented differences in fertility (farrowing rate, FR) and prolificacy (litter size, LS) (n = 7) in artificial insemination programs, using combined FR and LS, were categorized as High Fertile (HF, n = 4) or Low Fertile (LF, n = 3), and boars with Unknown Fertility (UF, n = 4) were tested for eventual epigenetical similarity with those fertility-proven. We identified 165,944 Single Nucleotide Polymorphisms (SNPs) that explained 14-15% of variance among selection lines. Between HF and LF individuals (n = 7, 4 HF and 3 LF), we identified 169 SNPs with p <= 0.00015, which explained 58% of the variance. For the epigenetic analyses, we considered fertility and period of ejaculate collection (late-summer and mid-autumn). Approximately three times more DMRs were observed in HF than in LF boars across these periods. Interestingly, UF boars were clearly clustered with one of the other HF or LF groups. The highest differences in DMRs between HF and LF experimental groups across the pig genome were located in the chr 3, 9, 13, and 16, with most DMRs being hypermethylated in LF boars. In both HF and LF boars, DMRs were mostly hypermethylated in late-summer compared to mid-autumn. Three overlaps were detected between SNPs (p <= 0.0005, n = 1318) and CpG sites within DMRs. In conclusion, fertility levels in breeding males including FR and LS can be discerned using methylome analyses. The findings in this biomedical animal model ought to be applied besides sire selection for andrological diagnosis of idiopathic sub/infertility.
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