SIRT1 gene methylation in sperm differs in rams with high and low fertility

Desislava Abadjieva, Nevqna Stancheva, Yordan Marchev, Elena Kistanova, Stanimir Yotov


Submitted 2020-07-02 | Accepted 2020-09-04 | Available 2020-12-01

Recently, more evidences of epigenetic impact on the male fertility, particularly on sperm DNA methylation have been reported. Data related to this issue in livestock males is still limited. The present study analyzed the DNA methylation status of the important gene for spermatogenesis, SIRT1, in ram sperm and its correspondence with semen quality and fertilizing ability. The ejaculates of 10 rams (5 rams - 1.5 years old, and 5 rams - 4 years old) from Synthetic Population Bulgarian Milk breed were evaluated and used for the artificial insemination of 174 ewes in breeding season. Two semen samples from each animal were used for DNA extraction followed by bisulfite conversion. The DNA methylation status of SIRT1 was detected through quantitative methylation-specific PCR using two sets of primers designed specifically for bisulfite-converted DNA sequences to attach methylated and unmethylated sites. On the base of age and conception rate the rams were divided in different groups. Data of semen quality, DNA methylation status of SIRT1 and reproductive performances of each group were statistically processed. Results showed a high average value of DNA methylation of SIRT1 in ram sperm (78.5±23.9%) and wide individual variability among investigated animals, with a coefficient of variation of 34.4%. The 1.5 years old animals tended to have a higher level of SIRT1 methylation than 4 years old animals. The rams in group with high fertilizing ability had significantly higher DNA methylation of SIRT1 in sperm than those with low fertilizing ability. In conclusion, results of this study provided evidence that the alteration of sperm SIRT1 methylation is associated with fertility performances of the rams and, probably, with their age.

Keywords: sperm DNA methylation, SIRT1, ram fertility


AHLAWAT, S. et al. (2019). Promoter methylation and expression analysis of Bvh gene in bulls with varying semen motility parameters. Theriogenology, 125, 152–156.

ASTON, K. I. et al. (2015). Aberrant sperm DNA methylation predicts male fertility status and embryo quality. Fertility and Sterility, 104, 1388–1397.

AX, R. L. et al. (2000). Semen evaluation. In: Hafez, B., Hafez, E. S. E. (Eds.), Reproduction in Farm Animals, 7 th ed. Lippincott Williams and Wilkins, Philadelphia, pp. 365-375.

BELL, E. L. et al. (2014). SirT1 is required in the male germ cell for differentiation and fecundity in mice. Development, 141(18), 3495-3504.

BOISSONNAS, C. C. et al. (2013). Epigenetic disorders and male subfertility. Fertility and Sterility, 99, 624–631.

CONGRAS, A. et al. (2014). Sperm DNA methylation analysis in swine reveals conserved and species-specific methylation patterns and highlights an altered methylation at theGNAS locus in infertile boars. Biology of Reproduction, 91(6), 137, 1–14.

COUSSENS, M. et al. (2008). Sirt1 Deficiency Attenuates Spermatogenesis and Germ Cell Function. PLoS ONE, 3(2), e1571.

DONKIN, I. and BARRES, R. (2018). Sperm epigenetics and influence of environmental factors. Molecular Metabolism, 14, 1–11.

ISLAM, S. et al. (2020). DNA hypermethylation of sirtuin 1 (SIRT1) caused by betel quid chewing—a possible predictive biomarker for malignant transformation. Clinical Epigenetics, 12, 12. JENKINS, T. G. et al. (2019). Age-associated sperm DNA methylation patterns do not directly persist trans-generationally. Epigenetics & Chromatin 12,(1), NA

JING, H. and LIN, H. (2015). Sirtuins in epigenetic regulation. Chemical Reviews, 115, 2350−2375.

KENNEDY, D. (2012). Sheep Reproduction Basics and Conception Rates.

KROPP, J. et al. (2017). Male fertility status is associated with DNA methylation signatures in sperm and transcriptomic profiles of bovine preimplantation embryos. BMC Genomics, 18, 280.

LAMBERT, S. et al. (2018). Spermatozoa DNA methylation patterns differ due to peripubertal age in bulls. Theriogenology, 106, 21–29.

LAQQAN, M. et al. (2017). Alterations in sperm DNA methylation patterns of oligospermic males. Reproductive Biology, 17, 396–400.

LIU, C. et al. (2017). Sirt1 regulates acrosome biogenesis by modulating autophagic flux during spermiogenesis in mice. Development, 144, 441-451.

MARTTILA, S. (2016). Ageing-associated Changes in Gene Expression and DNA Methylation. Academic dissertation. University of Tampere.

MCSWIGGIN, H. M. and O’DOHERTY, A. M. (2018). Epigenetic reprogramming during spermatogenesis and male factor infertility. Reproduction, 156, R9–R21.

MOLARO, A. et al. (2011). Sperm methylation profiles reveal features of epigenetic inheritance and evolution in primates. Cell, 146, 1029–1041.

NAYAK, K. et al. (2016). Epigenetic regulation of gene expression during spermatogenesis.

OAKES, C. C. et al. (2007). Developmental acquisition of genome-wide DNA methylation occurs prior to meiosis in male germ cells. Developmental Biology, 307, 368–379.

OLIVIER, W. J. (2014). Calculation of reproduction parameters. Info pack ref: AP 2014/032, Grootfontein Agricultural Development Institute.

PERRIER, J. P. et al. (2018). A multi-scale analysis of bull sperm methylome revealed both species peculiarities and conserved tissue-specific features. BMC Genomics, 19, 404.

RAHMAN S. and ISLAM, R. (2011). Mammalian Sirt1: insights on its biological functions. Cell Communication and Signaling, 9, 11.

SHARAFI, M. et al. (2017). Epigenetic modulation of ram sperm during cryopreservation. Reproduction in Domestic Animals 52(S3), 133.

SCHAGDARSURENGIN, U. and STEGER, K. (2016). Epigenetics in male reproduction: effect of paternal diet on sperm quality and offspring health. Nature Reviews Urology, 13, 584–595.

SHOJAEI SAADI, H. A. et al. (2017). Genome-wide analysis of sperm DNA methylation from monozygotic twin bulls. Reproduction, Fertility and Development, 29, 838–843.

TAKEDA, K. et al. 2019. Age-related changes in DNA methylation levels at CpG sites in bull spermatozoa and in vitro fertilization-derived blastocyst-stage embryos revealed by combined bisulfite restriction analysis. Journal of Reproduction and Development, 65, 305–312.

TANG, Q. et al. (2017). Idiopathic male infertility and polymorphisms in the DNA methyltransferase genes involved in epigenetic marking. Scientific Reports, 7, 11219.

TIBARY, A. et al. (2018). Ram and buck breeding soundness examination. Revue Marocaine des Sciences Agronomiques et Vétérinaires, 6(2), 241-255.

TOLIC, A. et al. (2019). Absence of PARP‐1 affects Cxcl12 expression by increasing DNA demethylation. Journal of Cellular and Molecular Medicine, 23, 2610–2618.

URDINGUIO, R. G. et al. (2015). Aberrant DNA methylation patterns of spermatozoa in men with unexplained infertility. Human Reproduction, 30,(5), 1014–1028.

VERMA, A. et al. (2014). Genome-wide profiling of sperm DNA methylation in relation to buffalo (Bubalus bubalis) bull fertility. Theriogenology, 82, 750–759.

WOLFFE, A. P. and GUSCHIN, D. (2000). Review: chromatin structural features and targets that regulate transcription. Journal of Structural Biology, 129, 102–122.

ZHOU, Y. et al. (2018). Comparative whole genome DNA methylation profiling of cattle sperm and somatic tissues reveals striking hypomethylated patterns in sperm. GigaScience, 7(5), giy039.


Full Text:



  • There are currently no refbacks.

Copyright (c) 2020 Acta Fytotechnica et Zootechnica

© Slovak University of Agriculture in Nitra, Faculty of Agrobiology and Food Resources