Skip to main content
Log in

Increase in the concentration of cytosolic-free calcium induced by human follicular fluid was decreased in single human spermatozoon with abnormal morphology

  • Original Article
  • Andrology
  • Published:
Reproductive Medicine and Biology

Abstract

Aim

The increase in the concentration of cytosolic-free calcium ([Ca2+]i) induced by follicular fluid or progesterone has been reported to promote an acrosome reaction and alternation in several motion parameters in human sperm (hyperactivation). We previously reported that populations of sperm in cell suspension obtained from infertile men with abnormal morphology exhibited lower mean peak progesterone-evoked [Ca2+]i compared with morphologically normal sperm using cell-suspension methods. In the present study, the change in [Ca2+]i in individual normally and abnormally shaped spermatozoa was compared.

Methods

The change in [Ca2+]i induced by human follicular fluid in individual spermatozoa with normal and abnormal morphology was compared using the fluorescent calciumsensitive dye fluo-3/AM. The spatial distribution of the increase in [Ca2+]i in single sperm was also investigated.

Results

The [Ca2+]i of normally shaped spermatozoa increased rapidly after the administration of human follicular fluid. The response reached a peak within 2–3 s and then slowly declined to a plateau phase. The baseline and peak fluorescence in spermatozoa with abnormal morphology was lower when compared with normal spermatozoa. The follicular-fluidinduced increase in [Ca2+]i (expressed as a percentage increase in [Ca2+]i over basal) in morphologically abnormal sperm was 39.2 ± 5.3% (n = 107, mean ± standard error), which was smaller than that of morphologically normal sperm (61.6 ± 5.7%, n = 100, P < 0.005) from seven healthy donors. The follicular-fluid-induced [Ca2+]i increases observed in sperm with morphologically abnormal mid-pieces (20.9 ± 4.3%, n = 12, P < 0.05) or tails (40.7 ± 6.0%, n = 92, P < 0.05) were lower than those of morphologically normal spermatozoa (61.6 ± 5.3%, n = 101). The follicular-fluid-induced [Ca2+]i increase of morphologically normal spermatozoa from infertile couples (35.1 ± 6.3%, n = 25, P < 0.05) was also found to be lower than that of morphologically normal spermatozoa from healthy donors.

Conclusion

The present study shows that spermatozoa with abnormal morphology in healthy donors have disorders of signal transduction, as do normally shaped sperm in men from infertile couples.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Liu DY, Baker HWG. Test of human sperm function and fertilization in vitro. Fertil Steril 1992; 58: 465–483.

    PubMed  CAS  Google Scholar 

  2. Osman RA, Andria ML, Jones AD et al. Steroid-induced exocytosis: the human sperm acrosome reaction. Biochem Biophys Res Commun 1989; 160: 828–833.

    Article  PubMed  CAS  Google Scholar 

  3. Baldi E, Casano R, Falsetti C et al. Intracellular calcium accumulation and responsiveness to progesterone in capacitating human spermatozoa. J Androl 1991; 12: 323–330.

    PubMed  CAS  Google Scholar 

  4. Meizel S, Turner KO. Progesterone acts at the plasma membrane of human sperm. Mol Cell Endocrinol 1991; 11: R1-R5.

    Article  Google Scholar 

  5. Bronson RA, Peresleni T, Golightly M. Progesterone promotes the acrosome reaction in capacitated human spermatozoa as judged by flow cytometry and CD46 staining. Mol Hum Reprod 1999; 5: 507–512.

    Article  PubMed  CAS  Google Scholar 

  6. Parat C, Serres C, Jouannet P. The acrosome reaction in human spermatozoa. Biol Cell 2000; 92: 255–266.

    Article  Google Scholar 

  7. Morales P, Llanos M, Gutierretz G et al. The acrosome reaction-inducing activity of individual human follicular fluid samples is highly variable and is related to the steroid content. Hum Reprod 1992; 7: 646–651.

    PubMed  CAS  Google Scholar 

  8. Saaranen ML, Calvo L, Dennison L et al. Acrosome reaction inducing activity in follicular fluid correlates with progesterone concentration but not with oocyte maturity or fertilizability. Hum Reprod 1993; 8: 1448–1454.

    PubMed  CAS  Google Scholar 

  9. Falsetti C, Baldi E, Krautz C et al. Decreased responsiveness to progesterone of spermatozoa in oligozoospermic patients. J Androl 1993; 14: 17–22.

    PubMed  CAS  Google Scholar 

  10. Tesarik J, Mendoza C. Defective function of a non-genomic progesterone receptor as a sole sperm anomaly in infertile patients. Fertil Steril 1992; 58: 793–797.

    PubMed  CAS  Google Scholar 

  11. Aitken RJ, Kelly RW. Analysis of the direct effects of prostaglandins on human sperm function. J Reprod Fertil 1985; 73: 139–146.

    PubMed  CAS  Google Scholar 

  12. Aitken RJ, Irvine S, Kelly RW. Significance of intracellular calcium and cytosolic adenosine 3′5′-monophosphate in the mechanisms by which prostaglandins influence human sperm function. J Reprod Fertil 1986; 77: 451–462.

    Article  PubMed  CAS  Google Scholar 

  13. Shimizu Y, Yorimitsu A, Maruyama Y et al. Prostaglandins induce calcium influx in human spermatozoa. Mol Hum Reprod 1998; 4: 555–561.

    Article  PubMed  CAS  Google Scholar 

  14. Enginsu ME, Damoulin JCM, Pieters MHEC et al. Predictive value of morphologically normal sperm concentration in the medium for in vitro fertilization. Int J Androl 1993; 16: 113–120.

    Article  PubMed  CAS  Google Scholar 

  15. Krausz C, Bonaccorsi L, Luconi M et al. Intracellular calcium increase and acrosome reaction in response to progesterone in human spermatozoa are correlated with in-vitro fertilization. Hum Reprod 1995; 10: 120–124.

    Article  PubMed  CAS  Google Scholar 

  16. Shimizu Y, Nord EP, Bronson RA. Progesterone-evoked increases in sperm [Ca2+]i correlate with the egg penetrating ability of sperm from fertile but not infertile men. Fertil Steril 1993; 60: 526–532.

    PubMed  CAS  Google Scholar 

  17. World Health Organization. Laboratory Manual for the Examination of Human Semen and Semen-Cervical Mucus Interactions, 4th edn. Cambridge University Press, Cambridge, 1999.

    Google Scholar 

  18. Waite LH, Tanzer ML. Specific colorimetric detection of odiphenols and 3,4-dihydroxyphenylalanine-containing peptides. Anal Biochem 1981; 111: 131–136.

    Article  PubMed  CAS  Google Scholar 

  19. Tesarik J, Carreras A, Mendoza C. Single cell analysis of tyrosine kinase dependent and independent Ca2+ fluxes in progesterone induced acrosome reaction. Mol Hum Reprod 1996; 2: 225–232.

    Article  PubMed  CAS  Google Scholar 

  20. Aitken RJ, Thatcher S, Glaser AF et al. Relative ability of modified version of the hamster oocyte penetration test, incorporating hyperosmotic medium or the ionophore A23187, to predict IVF outcome. Hum Reprod 1987; 2: 227–231.

    PubMed  CAS  Google Scholar 

  21. Pampiglione JS, Tan SL, Canpbell S. The use of stimulated acrosome reaction test as a test of fertilizing ability of human spermatozoa. Fertil Steril 1993; 59: 1280–1284.

    PubMed  CAS  Google Scholar 

  22. Harper CV, Publicover SJ. Reassessing the role of progesterone in fertilization-compartmentalized calcium signalling in human spermatozoa? Hum Reprod 2005; 20: 2675–2680.

    Article  PubMed  CAS  Google Scholar 

  23. Bedu-Addo K, Barratt CL, Kirkman-Brown JC et al. Patterns of [Ca2+](i) mobilization and cell response and cell response in human spermatozoa exposed to progesterone. Dev Biol 2007; 302: 324–332.

    Article  PubMed  CAS  Google Scholar 

  24. Aitken RJ, Buckingham DW, Irvine DS. The extragenomic action of progesterone on human spermatozoa: evidence for a ubiquitous response that is rapidly down-regulated. Endocrinol 1996; 137: 3999–4009.

    Article  CAS  Google Scholar 

  25. Oehninger S, Sueldo C, Blackmore P et al. Defective calcium influx and acrosome reaction (spontaneous and progesterone-induced) in spermatozoa of infertile men with severe teratozoospermia. Fertil Steril 1994; 61: 349–354.

    PubMed  CAS  Google Scholar 

  26. Shirakawa H, Miyazaki S. Spatiotemporal characterization of intracellular Ca2+ rise during the acrosome reaction of mammalian spermatozoa induced by zona pellucida. Dev Biol 1999; 208: 70–78.

    Article  PubMed  CAS  Google Scholar 

  27. Kirkman-Brown JC, Bray C, Stewart PM et al. Biphasic elevation of [Ca2+]i in individual human spermatozoa exposed to progesterone. Dev Biol 2000; 222: 326–335.

    Article  PubMed  CAS  Google Scholar 

  28. Watson PF, Plummer JM, Jones PS et al. Localization of intracellular calcium during the acrosome reaction in ram spermatozoa. Mol Reprod Dev 1995; 41: 513–520.

    Article  PubMed  CAS  Google Scholar 

  29. Florman HM. Sequential focal and global elevations of sperm intracellular Ca2+ are initiated by the zona pellucida during acrosomal exocytosis. Dev Biol 1994; 165: 152–164.

    Article  PubMed  CAS  Google Scholar 

  30. Tesarik J, Mendoza C, Moos J et al. Selective expression of progesterone receptor on the human sperm surface. Fertil Steril 1992; 58: 784–792.

    PubMed  CAS  Google Scholar 

  31. Liu DY, Baker HWG. Disordered acrosome reaction of sperm defect causing infertility with reduced sperm-ZP penetration and reduced fertilization in vitro. Hum Reprod 1994; 9: 1964–1700.

    Google Scholar 

  32. Liu DY, Baker HWG. Protein kinase C plays an important role in human zona pellucida-induced acrosome reaction. Mol Hum Reprod 1997; 3: 1037–1043.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yasufumi Shimizu.

About this article

Cite this article

Shimizu, Y., Minaguchi, R., Ishikawa, T. et al. Increase in the concentration of cytosolic-free calcium induced by human follicular fluid was decreased in single human spermatozoon with abnormal morphology. Reprod Med Biol 7, 143–149 (2008). https://doi.org/10.1111/j.1447-0578.2008.00211.x

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1111/j.1447-0578.2008.00211.x

Key words

Navigation