Journal of Assisted Reproduction and Genetics

, Volume 36, Issue 2, pp 267–275 | Cite as

Histone modification signatures in human sperm distinguish clinical abnormalities

  • Samantha B. SchonEmail author
  • Lacey J. Luense
  • Xiaoshi Wang
  • Marisa S. Bartolomei
  • Christos Coutifaris
  • Benjamin A. Garcia
  • Shelley L. BergerEmail author
Gamete Biology



Alternations to the paternal epigenome, specifically the components of sperm chromatin, can lead to infertility in humans and potentially transmit aberrant information to the embryo. One key component of sperm chromatin is the post-translational modification of histones (PTMs). We previously identified a comprehensive profile of histone PTMs in normozoospermic sperm; however, only specific histone PTMs have been identified in abnormal sperm by antibody-based approaches and comprehensive changes to histone PTM profiles remain unknown. Here, we investigate if sperm with abnormalities of total motility, progressive motility, and morphology have altered histone PTM profiles compared to normozoospermic sperm samples.


Discarded semen samples from 31 men with normal or abnormal semen parameters were analyzed for relative abundance of PTMs on histone H3 and H4 by “bottom-up” nano-liquid chromatography-tandem mass spectrometry.


Asthenoteratozoospermic samples (abnormal motility, forward progression, and morphology, n = 6) displayed overall decreased H4 acetylation (p = 0.001) as well as alterations in H4K20 (p = 0.003) and H3K9 methylation (p < 0.04) when compared to normozoospermic samples (n = 8). Asthenozoospermic samples (abnormal motility and progression, n = 5) also demonstrated decreased H4 acetylation (p = 0.04) and altered H4K20 (p = 0.005) and H3K9 methylation (p < 0.04). Samples with isolated abnormal progression (n = 6) primarily demonstrated decreased acetylation on H4 (p < 0.02), and teratozoospermic samples (n = 6) appeared similar to normozoospermic samples (n = 8).


Sperm samples with combined and isolated abnormalities of total motility, progressive motility, and morphology display distinct and altered histone PTM signatures compared to normozoospermic sperm. This provides evidence that alterations in histone PTMs may be important for normal sperm function and fertility.


Histones Post-translational modifications Paternal epigenetics Sperm Male infertility 



We would like to thank all of the staff at the Penn Fertility Care Andrology laboratory for their assistance, time, and effort in the de-identification of samples used in this study. In addition, we would like to thank The Penn Center for the Study of Epigenetics in Reproduction.


This work was funded by the National Institutes of Health Grants P50HD06817 (S.L.B./M.S.B./C.C/L.J.L), T32HD040135 (S.B.S/C.C), 5K12HD065257-07 (S.B.S), F32HD086939 (L.J.L.), and GM110174 (B.A.G).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Approval for this study was obtained from the University of Pennsylvania Institutional Review Board (Protocol 815929).

Informed consent

As all samples were discarded and de-identified, the University of Pennsylvania Institutional Review Board determined that this study was exempt from requiring informed consent.

Supplementary material

10815_2018_1354_MOESM1_ESM.docx (19 kb)
ESM 1 (DOCX 19 kb)


  1. 1.
    Stephen EH, Chandra A. Declining estimates of infertility in the United States: 1982–2002. Fertil Steril. 2006;86:516–23.CrossRefGoogle Scholar
  2. 2.
    Thonneau P, Marchand S, Tallec A, Ferial ML, Ducot B, Lansac J, et al. Incidence and main causes of infertility in a resident population (1,850,000) of three French regions (1988-1989). Hum Reprod. 1991;6:811–6.CrossRefGoogle Scholar
  3. 3.
    Cooper TG, Noonan E, von Eckardstein S, Auger J, Baker HWG, Behre HM, et al. World Health Organization reference values for human semen characteristics. Hum Reprod Update Oxford University Press. 2010;16:231–45.CrossRefGoogle Scholar
  4. 4.
    Larsen L. Computer-assisted semen analysis parameters as predictors for fertility of men from the general population. Hum Reprod. 2000;15:1562–7.CrossRefGoogle Scholar
  5. 5.
    Nallella KP, Sharma RK, Aziz N, Agarwal A. Significance of sperm characteristics in the evaluation of male infertility. Fertil Steril. 2006;85:629–34.CrossRefGoogle Scholar
  6. 6.
    Gunalp S, Onculoglu C, Gurgan T, Kruger TF, Lombard CJ. A study of semen parameters with emphasis on sperm morphology in a fertile population: an attempt to develop clinical thresholds. Hum Reprod. 2001;16:110–4.CrossRefGoogle Scholar
  7. 7.
    Vawda AI, Gunby J, Younglai EV. Andrology: semen parameters as predictors of in-vitro fertilization: the importance of strict criteria sperm morphology. Hum Reprod. 1996;11:1445–50.CrossRefGoogle Scholar
  8. 8.
    Verheyen G, Tournaye H, Staessen C, De Vos A, Vandervorst M, Van Steirteghem A. Controlled comparison of conventional in-vitro fertilization and intracytoplasmic sperm injection in patients with asthenozoospermia. Hum Reprod. 1999;14:2313–9.CrossRefGoogle Scholar
  9. 9.
    Kornberg RD. Structure of chromatin. Annu Rev Biochem. 1977;46:931–54.CrossRefGoogle Scholar
  10. 10.
    Zhao Y, Garcia BA. Comprehensive catalog of currently documented histone modifications. Cold Spring Harb Perspect Biol. 2015;7:a025064–22.CrossRefGoogle Scholar
  11. 11.
    Kouzarides T. Chromatin Modifications and Their Function. Cell. 2007;128:693–705.CrossRefGoogle Scholar
  12. 12.
    Tanphaichitr N, Sobhon P, Taluppeth N, Chalermisarachai P. Basic nuclear proteins in testicular cells and ejaculated spermatozoa in man. Exp Cell Res. 1978;117:347–56.CrossRefGoogle Scholar
  13. 13.
    Ward WS, Coffey DS. DNA packaging and organization in mammalian spermatozoa: comparison with somatic cells. Biol Reprod. 1991;44:569–74.CrossRefGoogle Scholar
  14. 14.
    Gannon JR, Emery BR, Jenkins TG, Carrell DT. The sperm epigenome: implications for the embryo. Adv Exp Med Biol New York, NY: Springer New York. 2014;791:53–66.CrossRefGoogle Scholar
  15. 15.
    Gatewood J, Cook G, Balhorn R, Bradbury E, Schmid C. Sequence-specific packaging of DNA in human sperm chromatin. Science. 1987;236:962–4.CrossRefGoogle Scholar
  16. 16.
    Zhang X. Sperm nuclear histone to protamine ratio in fertile and infertile men: evidence of heterogeneous subpopulations of spermatozoa in the ejaculate. J Androl. 2006;27:414–20.CrossRefGoogle Scholar
  17. 17.
    Chevaillier P, Mauro N, Feneux D, Jouannet P, David G. Anomalous protein complement of sperm nuclei in some infertile men. Lancet. 1987;2:806–7.CrossRefGoogle Scholar
  18. 18.
    de Mateo S, Ramos L, van der Vlag J, de Boer P, Oliva R. Improvement in chromatin maturity of human spermatozoa selected through density gradient centrifugation. Int J Androl. 2010;34:256–67.CrossRefGoogle Scholar
  19. 19.
    Carrell DT, Liu L. Altered protamine 2 expression is uncommon in donors of known fertility, but common among men with poor fertilizing capacity, and may reflect other abnormalities of spermiogenesis. J Androl. 2001;22:604–10.Google Scholar
  20. 20.
    Carrell DT, Emery BR, Hammoud S. Altered protamine expression and diminished spermatogenesis: what is the link? Hum Reprod Update. 2007;13:313–27.CrossRefGoogle Scholar
  21. 21.
    Aoki VW, Emery BR, Liu L, Carrell DT. Protamine levels vary between individual sperm cells of infertile human males and correlate with viability and DNA integrity. J Androl. 2006;27:890–8.CrossRefGoogle Scholar
  22. 22.
    Aoki V, Liu L, Jones K, Hatasaka H, Gibson M, Peterson C, et al. Sperm protamine 1/protamine 2 ratios are related to in vitro fertilization pregnancy rates and predictive of fertilization ability. Fertil Steril. 2006;86:1408–15.CrossRefGoogle Scholar
  23. 23.
    Hammoud SS, Nix DA, Zhang H, Purwar J, Carrell DT, Cairns BR. Distinctive chromatin in human sperm packages genes for embryo development. Nature Nature Publishing Group. 2009;460:473–8.CrossRefGoogle Scholar
  24. 24.
    Arpanahi A, Brinkworth M, Iles D, Krawetz SA, Paradowska A, Platts AE, et al. Endonuclease-sensitive regions of human spermatozoal chromatin are highly enriched in promoter and CTCF binding sequences. Genome Res. 2009;19:1338–49.CrossRefGoogle Scholar
  25. 25.
    Brykczynska U, Hisano M, Erkek S, Ramos L, Oakeley EJ, Roloff TC, et al. Repressive and active histone methylation mark distinct promoters in human and mouse spermatozoa. Nat Struct Mol Biol. 2010;17:679–87.CrossRefGoogle Scholar
  26. 26.
    Jung YH, Sauria MEG, Lyu X, Cheema MS, Ausió J, Taylor J, et al. Chromatin states in mouse sperm correlate with embryonic and adult regulatory landscapes. Cell Rep. ElsevierCompany. 2017;18:1366–82.CrossRefGoogle Scholar
  27. 27.
    Samans B, Yang Y, Krebs S, Sarode GV, Blum H, Reichenbach M, et al. Uniformity of nucleosome preservation pattern in mammalian sperm and its connection to repetitive DNA elements. Dev Cell. 2014;30:23–35.CrossRefGoogle Scholar
  28. 28.
    Carone BR, Hung J-H, Hainer SJ, Chou M-T, Carone DM, Weng Z, et al. High-resolution mapping of chromatin packaging in mouse embryonic stem cells and sperm. Dev Cell 2014;1–20. Elsevier Inc.Google Scholar
  29. 29.
    Yuan ZF, Arnaudo AM, Garcia BA. Mass spectrometric analysis of histone proteoforms. Annu Rev Anal Chem (Palo Alto Calif). 2014;7:113–28.CrossRefGoogle Scholar
  30. 30.
    Luense LJ, Wang X, Schon SB, Weller AH, Shiao EL, Bryant JM, et al. Comprehensive analysis of histone post-translational modifications in mouse and human male germ cells. Epigenetics Chromatin BioMed Central. 2016;9:1–15.CrossRefGoogle Scholar
  31. 31.
    Kruger TF, Acosta AA, Simmons KF, Swanson RJ, Matta JF, Oehninger S. Predictive value of abnormal sperm morphology in in vitro fertilization. Fertil Steril. 1988;49:112–7.CrossRefGoogle Scholar
  32. 32.
    Shechter D, Dormann HL, Allis CD, Hake SB. Extraction, purification and analysis of histones. Nat Protoc. 2007;2:1445–57.CrossRefGoogle Scholar
  33. 33.
    Lin S, Garcia BA. Examining histone posttranslational modification patterns by high-resolution mass spectrometry. Methods Enzymol. 2012;512:3–28.Google Scholar
  34. 34.
    Govin J, Lestrat C, Caron C, Pivot-Pajot C, Rousseaux S, Khochbin S. Histone acetylation-mediated chromatin compaction during mouse spermatogenesis. Ernst Schering Res Found Workshop. 2006;57:155–72.Google Scholar
  35. 35.
    Boissonnas CC, Jouannet P, Jammes H. Epigenetic disorders and male subfertility. Fertil Steril. 2013;99:624–31.CrossRefGoogle Scholar
  36. 36.
    Rathke C, Baarends WM, Awe S, Renkawitz-Pohl R. Chromatin dynamics during spermiogenesis. Biochim Biophys Acta. 1839;2014:155–68.Google Scholar
  37. 37.
    Goudarzi A, Shiota H, Rousseaux S, Khochbin S. Genome-scale acetylation-dependent histone eviction during spermatogenesis. J Mol Biol Elsevier Ltd. 2014;426:3342–9.CrossRefGoogle Scholar
  38. 38.
    Dada R, Kumar M, Jesudasan R, Fernández JL, Gosálvez J, Agarwal A. Epigenetics and its role in male infertility. J Assist Reprod Genet. 2012;29:213–23.CrossRefGoogle Scholar
  39. 39.
    Faure AK. Misregulation of histone acetylation in Sertoli cell-only syndrome and testicular cancer. Mol Hum Reprod. 2003;9:757–63.CrossRefGoogle Scholar
  40. 40.
    Sonnack V, Failing K, Bergmann M, Steger K. Expression of hyperacetylated histone H4 during normal and impaired human spermatogenesis. Andrologia. 2002;34:384–90.CrossRefGoogle Scholar
  41. 41.
    van Nuland R, Gozani O. Histone H4 lysine 20 (H4K20) methylation, expanding the signaling potential of the proteome one methyl moiety at a time. Mol Cell Proteomics. 2016;15:755–64.CrossRefGoogle Scholar
  42. 42.
    Jørgensen S, Schotta G, Sørensen CS. Histone H4 lysine 20 methylation: key player in epigenetic regulation of genomic integrity. Nucleic Acids Res Oxford University Press. 2013;41:2797–806.CrossRefGoogle Scholar
  43. 43.
    Grewal SIS, Jia S. Heterochromatin revisited. Nat Rev Genet. 2007;8:35–46.CrossRefGoogle Scholar
  44. 44.
    Okada Y, Scott G, Ray MK, Mishina Y, Zhang Y. Histone demethylase JHDM2A is critical for Tnp1 and Prm1 transcription and spermatogenesis. Nature. 2007;450:119–23.CrossRefGoogle Scholar
  45. 45.
    Peters A, O'Carroll D, Scherthan H, Mechtler K. Loss of the Suv39h histone methyltransferases impairs mammalian heterochromatin and genome stability. Cell. 2001;107:323–37.CrossRefGoogle Scholar
  46. 46.
    van de Werken C, van der Heijden GW, Eleveld C, Teeuwssen M, Albert M, Baarends WM, et al. Paternal heterochromatin formation in human embryos is H3K9/HP1 directed and primed by sperm-derived histone modifications. Nat Commun. 2014;5:5868.CrossRefGoogle Scholar
  47. 47.
    Steilmann C, Paradowska A, Bartkuhn M, Vieweg M, Schuppe HC, Bergmann M, et al. Presence of histone H3 acetylated at lysine 9 in male germ cells and its distribution pattern in the genome of human spermatozoa. Reprod Fertil Dev. 2011;23:997–15.CrossRefGoogle Scholar
  48. 48.
    Vieweg M. Methylation analysis of histone H4K12ac-associated promoters in sperm of healthy donors and subfertile patients. Clin Epigenetics BioMed Central. 2015;7:1–17.CrossRefGoogle Scholar
  49. 49.
    Hammoud SS, Nix DA, Hammoud AO, Gibson M, Cairns BR, Carrell DT. Genome-wide analysis identifies changes in histone retention and epigenetic modifications at developmental and imprinted gene loci in the sperm of infertile men. Hum Reprod. 2011;26:2558–69.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Samantha B. Schon
    • 1
    • 2
    Email author
  • Lacey J. Luense
    • 3
    • 4
  • Xiaoshi Wang
    • 4
    • 5
  • Marisa S. Bartolomei
    • 3
    • 4
  • Christos Coutifaris
    • 1
  • Benjamin A. Garcia
    • 4
    • 5
  • Shelley L. Berger
    • 3
    • 4
    Email author
  1. 1.Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and GynecologyUniversity of PennsylvaniaPhiladelphiaUSA
  2. 2.Division of Reproductive Endocrinology & Infertility, Department of Obstetrics and GynecologyUniversity of Michigan Medical SchoolAnn ArborUSA
  3. 3.Department of Cell and Developmental Biology, Perelman School of Medicine, 9-125 Smilow Center for Translational ResearchUniversity of PennsylvaniaPhiladelphiaUSA
  4. 4.Epigenetics Institute, Perelman School of Medicine, 9-125 Smilow Center for Translational ResearchUniversity of PennsylvaniaPhiladelphiaUSA
  5. 5.Department of Biochemistry and Molecular Biophysics, Perelman School of Medicine, 9-125 Smilow Center for Translational ResearchUniversity of PennsylvaniaPhiladelphiaUSA

Personalised recommendations