Skip to main content

Human Cervical Mucus Can Act in Vitro as a Selective Barrier Against Spermatozoa Carrying Fragmented DNA and Chromatin Structural Abnormalities


Purpose : We have carried out experiments to determine if human cervical mucus can act as an in vitro selective barrier against spermatozoa morphologically normal that carry genetic structural abnormalities.

Methods : Sperm chromatin abnormalities have been evaluated by Chromomycin A3 and “endogenous” nick translation.

Results : The data obtained have shown that spermatozoa possessing higher levels of DNA protamination are more proficient in crossing the cervical mucus barrier. Moreover, the levels of positivity to endogenous nick translation treatment was practically zero in such spermatozoa.

Conclusions : We suggest that sperm penetration of cervical mucus could be used to select sperm preparations free of fragmented DNA or chromatin structural abnormalities for assisted reproduction.

This is a preview of subscription content, access via your institution.


  1. 1.

    Barrat, CLR, Osborn, JC, Harrison, PE, Monks, N, Dumphy, BC, Lenton, EA, Cooke, ID: The hypo-osmotic swelling test and sperm mucus penetration test in determining fertilization of the human oocyte. Hum Reprod 1989;4:430-434

    Google Scholar 

  2. 2.

    Eggert-Kruse, W, Schwarz, H, Rohr, G, Demirakca, T, Tilgen, W, Runnebaum, B: Sperm morphology assessment using strict criteria and male fertility under in-vivo conditions of conception. Hum Reprod 1996;11:139-146

    Google Scholar 

  3. 3.

    Abu-Heija, AT, Fleming, R, Jamieson, ME, Yates, RW, Coutts, JR: The predictive value of the sperm-cervical mucus interaction test on the outcome of in vitro fertilization and ovulation induction combined with intrauterine insemination. Aust NZJ Obstet Gynaecol 1997;37:232-234

    Google Scholar 

  4. 4.

    Baker, HW, Clarke, GN, Hudson, B, McBain, J-C, McGowan, MP, Pepperell, RJ: Treatment of sperm autoimmunity in men. Clin Reprod Fertil 1983;2:55-71

    Google Scholar 

  5. 5.

    Katz, DF, Overstreet, JW, Hanson, FW: A new quantitative test for sperm penetration into cervical mucus. Fertil Steril 1980;33:179-186

    Google Scholar 

  6. 6.

    Katz, DF, Morales, P, Samuels, SJ, Overstreet, JW: Mechanisms of filtration of morphologically abnormal human sperm by cervical mucus. Fertil Steril 1990;54:513-516

    Google Scholar 

  7. 7.

    Katz, DF: Human cervical mucus: Research update. Am J Obstet Gynecol 1991;165:1984-1986

    Google Scholar 

  8. 8.

    Aitken, RJ, Sutton, M, Warner, P, Richardson, DW: Relationship between the movement characteristics of human spermatozoa and their ability to penetrate cervical mucus and zona-free hamster oocytes. J Reprod Fertil 1985;73:441-449

    Google Scholar 

  9. 9.

    Eggert-Kruse, W, Reimann-Andersen, J, Rohr, G, Pohl, S, Tilgen, W, Runnebaum, B: Clinical relevance of sperm morphology assessment using strict criteria and relationship with sperm-mucus interaction in vivo and in vitro. Fertil Steril 1995;63:612-624

    Google Scholar 

  10. 10.

    Clarke, GN: A simplified quantitative cervical mucus penetration test. Hum Reprod 1997;12:1184-1187

    Google Scholar 

  11. 11.

    Clarke, GN, Garrett, C, Baker, HW: Quantitative sperm mucus penetration: Modified formulae for calculating penetration efficiency. Hum Reprod 1998;13:1255-1259

    Google Scholar 

  12. 12.

    Evenson, DP, Darzynkiewicz, Z, Melamed, MR: Relation of mammalian sperm chromatin heterogeneity to fertility. Science 1980;210:1131-1133

    Google Scholar 

  13. 13.

    Evenson, DP, Jost, LK, Marshall, D, Zinaman, MJ, Clegg, E, Purvis, K, de Angelis, P, Claussen, OP: Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic. Hum Reprod 1999;3:1039-1049

    Google Scholar 

  14. 14.

    Bianchi, PG, Manicardi, GC, Bizzaro, D, Bianchi, U, Sakkas, D: Effect of deoxyribonucleic acid protamination on fluorochrome staining and in situ nick translation of murine and human nature spermatozoa. Biol Reprod 1993;49:1083-1088

    Google Scholar 

  15. 15.

    Manicardi, GC, Bianchi, PG, Pantano, S, Azzoni, P, Bizzaro, D, Bianchi, U, Sakkas, D: Presence of endogenous nicks in DNA of ejaculated human spermatozoa and its relationship to Chromomycin A3 accessibility. Biol Reprod 1995;52:864-867

    Google Scholar 

  16. 16.

    Manicardi, GC, Tombacco, A, Bizzaro, D, Bianchi, U, Bianchi, PG, Sakkas, D: DNA strand breaks in ejaculated human spermatozoa: Comparison of susceptibility to the nick translation and terminal transferase assays. Histochem J 1998;30:33-39

    Google Scholar 

  17. 17.

    Sakkas, D, Urner, F, Bianchi, PG, Bizzaro, D, Wagner, I, Jaquenoud, N, Manicardi, GC, Campana, A: Sperm chromatin abnormalities can influence decondensation after intracytoplasmic sperm injection. Hum Reprod 1996;11:837-843

    Google Scholar 

  18. 18.

    Sakkas, D, Urner, F, Bizzaro, D, Bianchi, PG, Manicardi, GC, Shoukir, Y, Campana, A: Sperm nuclear DNA damage and altered chromatin structure: Effect on fertilization and embryo development. Hum Reprod 1998;4(Suppl):11-19

    Google Scholar 

  19. 19.

    Qiu, J, Hales, BF, Robaire, B: Damage to rat spermatozoal DNA after chronic cyclophosphamide exposure. Biol Reprod 1995;53:1465-1473

    Google Scholar 

  20. 20.

    Qiu, J, Hales, BF, Robaire, B: Effects of chronic low-dose cyclophosphamide exposure on the nuclei of rat spermatozoa. Biol Reprod 1995;52:33-40

    Google Scholar 

  21. 21.

    Lopes, S, Sun, JG, Juriscova, A, Meriano, J, Casper, RF: Sperm deoxyribonucleic acid fragmentation is increased in poor quality sperm samples and correlated with failed fertilization in intracytoplasmic sperm injection. Fertil Steril 1998;69:528-532

    Google Scholar 

  22. 22.

    Bianchi, PG, Manicardi, GC, Bizzaro, D, Campana, A, Bianchi, U, Sakkas, D: Use of the guanine-cytosine (GC) specific fluorochrome Chromomycin A3, as an indicator of poor sperm morphology. J Assist Reprod Genet 1996;13:246-250

    Google Scholar 

  23. 23.

    Bizzaro, D, Manicardi, GC, Bianchi, PG, Bianchi, U, Mariethoz, E, Sakkas, D: In situ competition between protamine and fluorochromes for sperm DNA. Mol Hum Reprod 1998;4:127-132

    Google Scholar 

  24. 24.

    Gao, X, Patel, DJ: Chromomycin dimer-DNA oligomer complex. Sequence selectivity and divalent cation specificity. Biochemistry 1990;29:10940-10956

    Google Scholar 

  25. 25.

    Gao, X, Mirau, P, Patel, DJ: Structure refinement of the chromomycin dimer-DNA oligomer complex in solution. J Mol Biol 1992;223:259-279

    Google Scholar 

  26. 26.

    Goodsell, DS, Kopka, ML, Cascio, D, Dickerson, RE: Crystal structure of CATGGCCATG and its implication for A-tract bending models. Proc Natl Acad Sci USA 1993;90:2930-2934

    Google Scholar 

  27. 27.

    Moghissi, KS, Lyner, FN: Studies in the human cervical mucus: Mucoids and their relation to sperm penetration. Fertil Steril 1970;21:234-239

    Google Scholar 

  28. 28.

    Barros, C, Vigil, P, Herrera, E, Guadarrama, A, Bustos-Obregon, E: In vitro interaction between human spermatozoa and human cervical mucus. Micr Elect Biol Cell 1983;7:13-19

    Google Scholar 

  29. 29.

    Vigil, P, Valdez, E: Bases cientìficas de la planificaciòn natural de la familia. Actual Ginec Obstet 1989;3:59-67

    Google Scholar 

  30. 30.

    Vigil, P, Perez, A, Neira, J, Morales, P: Post-partum cervical mucus: Biological and rheological properties. Hum Reprod 1991;6:475-479

    Google Scholar 

  31. 31.

    Fredricsson, B, Björk, G: Morphology of postcoital spermatozoa in the cervical secretion and its clinical significance. Fertil Steril 1977;28:841-845

    Google Scholar 

  32. 32.

    Hanson, FW, Overstreet, JW: The interaction of human spermatozoa with cervical mucus in vivo. Am J Obstet Gynaecol 1981;140:173-178

    Google Scholar 

  33. 33.

    Foresta, C, Zorzi, M, Rossato, M, Varotto, A: Sperm nuclear instability and staining with aniline blue: Abnormal persistence of histones in spermatozoa in infertile men. Int J Androl 1992;15:330-337

    Google Scholar 

  34. 34.

    Bianchi, PG, Manicardi, GC, Urner, F, Campana, A, Sakkas, D: Chromatin packaging and morphology in ejaculated human spermatozoa: Evidence of hidden anomalies in normal spermatozoa. Mol Hum Reprod 1996;2:139-144

    Google Scholar 

  35. 35.

    Hughes, CM, Lewis, SEM, McClavey-Martin, VJ, Tomphson, W: The effect of antioxidant supplementation during Percol preparation on human sperm DNA integrity. Mol HumReport 1998;13:1240-1247.

    Google Scholar 

Download references

Author information



Rights and permissions

Reprints and Permissions

About this article

Cite this article

Bianchi, P., De Agostini, A., Fournier, J. et al. Human Cervical Mucus Can Act in Vitro as a Selective Barrier Against Spermatozoa Carrying Fragmented DNA and Chromatin Structural Abnormalities. J Assist Reprod Genet 21, 97–102 (2004).

Download citation

  • Chromatin abnormalities
  • Chromomycin A3
  • human cervical mucus
  • in situ nick translation
  • spermatozoa