Ukr.Biochem.J. 2018; Volume 90, Issue 1, Jan-Feb, pp. 48-57


Responsiveness to progesterone and potassium channel blockers 4-aminopyridine, tetraethylammonium and free Ca(2+) contentration in spermatozoa of patients with oligozoospermia/leucocytospermia

R. V. Fafula1, G. V. Danylovych2, A. S. Besedina1, O. V. Melnyk1, Z. D. Vorobets1

1Danylo Halytsky Lviv National Medical University, Ukraine;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;

The present study was undertaken to evaluate [Ca2+]i signals that occur in human sperm cells exposed in vitro to three diverse compounds; progesterone, 4-aminopyridine (a highly effective inducer of hyperactivation in human sperm) and tetraethylammonium. The [Ca2+]i reached after the extracellular calcium treatment was always higher in normozoospermic samples pretreated with progesterone than in pathozoospermic samples pretreated with progesterone. There were no changes in calcium signal in spermatozoa pretreated with progesterone from patients with oligozoospermia and leucocytospermia compared to control samples (without progesterone). [Ca2+]i was always higher in pathozoospermic samples without 4-aminopyridine and always lower in pathozoospermic samples with 4-aminopyridine compared to these values in normozoospermic men. The 2 mM extracellular calcium administration to spermatozoa pretreated with tetraethylammonium did not result in a detectable increase in [Ca2+]i in normo- and pathozoospermic samples. The mechanisms of progesterone-dependent activation of the Ca2+-entry and the functioning of the voltage gated Ca2+-channels of plasmalemma are disturbed in pathologies – there was no increase in the Ca2+ level in the conditions of K+-depolarization (in the presence of inhibitors of K+-channels).

Keywords: , , ,


  1. Jung JH, Kim MH, Kim J, Baik SK, Koh SB, Park HJ, Seo JT. Treatment of Leukocytospermia in Male Infertility: A Systematic Review. World J Mens Health. 2016 Dec;34(3):165-172. PubMed, PubMedCentral, CrossRef
  2. Publicover S, Harper CV, Barratt C. [Ca2+]i signalling in sperm – making the most of what you’ve got. Nat Cell Biol. 2007 Mar;9(3):235-42. PubMed, CrossRef
  3. Publicover SJ, Giojalas LC, Teves ME, de Oliveira GS, Garcia AA, Barratt CL, Harper CV. Ca2+ signalling in the control of motility and guidance in mammalian sperm. Front Biosci. 2008 May 1;13:5623-37. PubMed, CrossRef
  4. Espino J, Mediero M, Lozano GM, Bejarano I, Ortiz A, García JF, Pariente JA, Rodríguez AB. Reduced levels of intracellular calcium releasing in spermatozoa from asthenozoospermic patients. Reprod Biol Endocrinol. 2009 Feb 6;7:11. PubMed, PubMedCentral, CrossRef
  5. Marquez B, Ignotz G, Suarez SS. Contributions of extracellular and intracellular Ca2+ to regulation of sperm motility: Release of intracellular stores can hyperactivate CatSper1 and CatSper2 null sperm. Dev Biol. 2007 Mar 1;303(1):214-21.  PubMed, PubMedCentral, CrossRef
  6. Eisenbach M, Giojalas LC. Sperm guidance in mammals – an unpaved road to the egg. Nat Rev Mol Cell Biol. 2006 Apr;7(4):276-85. PubMed, CrossRef
  7. Felix R. Molecular physiology and pathology of Ca2+-conducting channels in the plasma membrane of mammalian sperm. Reproduction. 2005 Mar;129(3):251-62. PubMed, CrossRef
  8. Ho K, Wolff CA, Suarez SS. CatSper-null mutant spermatozoa are unable to ascend beyond the oviductal reservoir. Reprod Fertil Dev. 2009;21(2):345-50. PubMed, CrossRef
  9. Al-Dossary AA, Strehler EE, Martin-Deleon PA. Expression and secretion of plasma membrane Ca2+-ATPase 4a (PMCA4a) during murine estrus: association with oviductal exosomes and uptake in sperm. PLoS One. 2013 Nov 14;8(11):e80181. PubMed, PubMedCentral, CrossRef
  10. Krasznai Z, Krasznai ZT, Morisawa M, Bazsáné ZK, Hernádi Z, Fazekas Z, Trón L, Goda K, Márián T. Role of the Na+/Ca2+ exchanger in calcium homeostasis and human sperm motility regulation. Cell Motil Cytoskeleton. 2006 Feb;63(2):66-76. PubMed, CrossRef
  11. Ho HC, Suarez SS. Characterization of the intracellular calcium store at the base of the sperm flagellum that regulates hyperactivated motility. Biol Reprod. 2003 May;68(5):1590-6. PubMed, CrossRef
  12. Lawson C, Dorval V, Goupil S, Leclerc P. Identification and localisation of SERCA 2 isoforms in mammalian sperm. Mol Hum Reprod. 2007 May;13(5):307-16.  PubMed, CrossRef
  13. Harper C, Wootton L, Michelangeli F, Lefièvre L, Barratt C, Publicover S. Secretory pathway Ca2+-ATPase (SPCA1) Ca(2)+ pumps, not SERCAs, regulate complex [Ca2+]i signals in human spermatozoa. J Cell Sci. 2005 Apr 15;118(Pt 8):1673-85. PubMed, CrossRef
  14. Gunaratne HJ, Vacquier VD. Evidence for a secretory pathway Ca2+-ATPase in sea urchin spermatozoa. FEBS Lett. 2006 Jul 10;580(16):3900-4.
    PubMed, CrossRef
  15. Costello S, Michelangeli F, Nash K, Lefievre L, Morris J, Machado-Oliveira G, Barratt C, Kirkman-Brown J, Publicover S. Ca2+-stores in sperm: their identities and functions. Reproduction. 2009 Sep;138(3):425-37. PubMed, PubMedCentral, CrossRef
  16. Amaral A, Lourenço B, Marques M, Ramalho-Santos J. Mitochondria functionality and sperm quality. Reproduction. 2013 Oct 1;146(5):R163-74.  PubMed, CrossRef
  17. Kolomiets OV, Danylovych YuV, Danylovych HV, Kosterin SO. Ca2+/H+-exchange in myometrium mitochondria. Ukr Biochem J. 2014; 86(3): 41-48. (In Ukrainian). PubMed, CrossRef
  18. Meseguer M, Garrido N, Martínez-Conejero JA, Simón C, Pellicer A, Remohí J. Relationship between standard semen parameters, calcium, cholesterol contents, and mitochondrial activity in ejaculated spermatozoa from fertile and infertile males. J Assist Reprod Genet. 2004 Dec;21(12):445-51. PubMed, PubMedCentral, CrossRef
  19. Munuce MJ, Quintero I, Caille AM, Ghersevich S, Berta CL. Comparative concentrations of steroid hormones and proteins in human peri-ovulatory peritoneal and follicular fluids. Reprod Biomed Online. 2006 Aug;13(2):202-7.  PubMed, CrossRef
  20. Teves ME, Barbano F, Guidobaldi HA, Sanchez R, Miska W, Giojalas LC. Progesterone at the picomolar range is a chemoattractant for mammalian spermatozoa. Fertil Steril. 2006 Sep;86(3):745-9. PubMed, CrossRef
  21. Harper CV, Kirkman-Brown JC, Barratt CL, Publicover SJ. Encoding of progesterone stimulus intensity by intracellular [Ca2+] ([Ca2+]i) in human spermatozoa. Biochem J. 2003 Jun 1;372(Pt 2):407-17. PubMed, PubMedCentral, CrossRef
  22. Strünker T, Goodwin N, Brenker C, Kashikar ND, Weyand I, Seifert R, Kaupp UB. The CatSper channel mediates progesterone-induced Ca2+ influx in human sperm. Nature. 2011 Mar 17;471(7338):382-6.  PubMed, CrossRef
  23. Bedu-Addo K, Barratt CL, Kirkman-Brown JC, Publicover SJ. Patterns of [Ca2+](i) mobilization and cell response in human spermatozoa exposed to progesterone. Dev Biol. 2007 Feb 1;302(1):324-32. PubMed, CrossRef
  24. Petersen OH, Sutton R, Criddle DN. Failure of calcium microdomain generation and pathological consequences. Cell Calcium. 2006 Nov-Dec;40(5-6):593-600. PubMed, CrossRef
  25. Alexander SP, Mathie A, Peters JA. Guide to receptors and channels, 2nd edition. Br J Pharmacol. 2006 Mar;147(S3):S1-168. PubMed, PubMedCentral, CrossRef
  26. Bedu-Addo K, Costello S, Harper C, Machado-Oliveira G, Lefievre L, Ford C, Barratt C, Publicover S. Mobilisation of stored calcium in the neck region of human sperm – a mechanism for regulation of flagellar activity. Int J Dev Biol. 2008;52(5-6):615-26.  PubMed, CrossRef
  27. Navarro B, Kirichok Y, Clapham DE. KSper, a pH-sensitive K+ current that controls sperm membrane potential. Proc Natl Acad Sci USA. 2007 May 1;104(18):7688-92.  PubMed, PubMedCentral, CrossRef
  28. Xia J, Ren D. Egg coat proteins activate calcium entry into mouse sperm via CATSPER channels. Biol Reprod. 2009 Jun;80(6):1092-8.  PubMed, PubMedCentral, CrossRef
  29. Alasmari W, Costello S, Correia J, Oxenham SK, Morris J, Fernandes L, Ramalho-Santos J, Kirkman-Brown J, Michelangeli F, Publicover S, Barratt CL. Ca2+ signals generated by CatSper and Ca2+ stores regulate different behaviors in human sperm. J Biol Chem. 2013 Mar 1;288(9):6248-58. PubMed, PubMedCentral, CrossRef
  30. Parodi J, Navarrete P, Marconi M, Gutiérrez RS, Martínez-Torres A, Mejías FR. Tetraethylammonium-sensitive K(+) current in the bovine spermatozoa and its blocking by the venom of the Chilean Latrodectus mactans. Syst Biol Reprod Med. 2010 Feb;56(1):37-43. PubMed, CrossRef
  31. Kasatkina LA. 4-Аminopyridine sequesters intracellular Ca2+ which triggers exocytosis in excitable and non-excitable cells. Sci Rep. 2016 Oct 5;6:34749. PubMed, PubMedCentral, CrossRef
  32. Giojalas LC, Iribarren P, Molina R, Rovasio RA, Estofán D. Determination of human sperm calcium uptake mediated by progesterone may be useful for evaluating unexplained sterility. Fertil Steril. 2004 Sep;82(3):738-40. PubMed, CrossRef

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