Role of Calcium and Calcium Channels in Progesterone Induced Acrosome Reaction in Caprine Spermatozoa

  • Somanath, P.R. ;
  • Gandhi, K.K.
  • Received : 2001.09.13
  • Accepted : 2002.02.15
  • Published : 2002.07.01


There are several physiological and pharmacological evidences indicating that opening of voltage dependent $Ca^{2+}$ channels play a critical role in induction of acrosome reaction in mammalian sperm. We determined the intracellular free $Ca^{2+}$ concentration in ejaculated goat sperm using a fluorescent, $Ca^{2+}$-specific probe, Fura2/AM, after the suspension of sperm in KRB medium, capable of sustaining capacitation and the acrosome reaction. We used nifedipine, D-600 and diltiazem, the $Ca^{2+}$ channel antagonists belonging to the classes of dihydropyridines, phenylalkylamines and benzothiazepines, to investigate the possibility that L-type voltage gated $Ca^{2+}$ channels play a role in the progesterone-stimulated exocytotic response. Progesterone promoted a rise in intracellular $Ca^{2+}$ in goat sperm and addition of nifedipine (100 nM) just prior to progesterone induction, significantly inhibited both intracellular $Ca^{2+}$ rise and exocytosis suggesting that $Ca^{2+}$ channels are involved in the process. However, the intracellular $Ca^{2+}$ increase during the process of capacitation was not affected with the addition of nifedipine suggesting a role of focal channel for $Ca^{2+}$ during capacitation. Studies using monensin and nigericin, two monovalent cation ionophores showed that an influx of $Na^+$ also may play a role in the opening of $Ca^{2+}$ channels. These results strongly suggests that the entry of $Ca^{2+}$ channels with characteristics similar to those of L-type, voltage-sensitive $Ca^{2+}$ channels found in cardiac and skeletal muscle, is a crucial step in the sequence of events leading to progesterone induced acrosome reaction in goat sperm.


Goat Sperm;Nifedipine;Monensin;Nigericin;$Ca^{2+}$;Channels;Fura 2


  1. Blackmore, P. F. and F. A. Lattanzio. 1991. Cell surface localization of a novel non-genomic progesterone receptor on the head of human sperm. Biochem. Biophys. Res. Commun. 181:331-336.
  2. Gonzalez-Martinez, M. T., B. E. Galindo, L. de De La Torre, O. Zapata, E. Rodriguez, H. M. Florman and A. Darszon. 2001. A sustained increase in intracellular $Ca^{2+}$ is required for the acrosome reaction in sea urchin sperm. Dev. Biol. 236:220-229.
  3. Ijima, T., S. Ciani and S. Hagawara. 1986. Effects of the external pH on Ca2+ channels: Experimental studies and theoretical considerations using a two-site, two-ion model. Proc. Natl. Acad. Sci. USA, 83:654-658.
  4. Meizel, S., M. Pillai, E. Diaz-Perez and P. Thomas. 1990. Initiation of the human sperm acrosome reaction by components of human follicular fluid and cumulus secretions including steroids. In: 'Fertilization in Mammals' (Ed. B. D. Bavister, J. M. Cummins, E. R. S. Roldan). Serona Symposia, USA, Norwell MA, pp. 205-222.
  5. Osman, R. A., M. L. Andria, A. D. Jones and S. Meizel. 1989. Steroid induced exocytosis: The human sperm acrosome reaction. Biochem. Biophys. Res. Commun. 160:828-833.
  6. Reuter, H. 1983. Calcium channel modulation by neurotransmitters, enzymes and drugs. Nature 301:569-574.
  7. Sabeur, K., D. P. Edwards and S. Meizel. 1996. Human sperm plasma membrane progesterone receptor(s) and the acrosome reaction. Biol. Reprod. 54:993-1001.
  8. Shi, Q. X. and E. R. S. Roldan. 1995. Evidence that a GABAA-like receptor is involved in progesterone-induced acrosomal exocytosis in mouse spermatozoa. Biol. Reprod. 52:373-381.
  9. Somanath, P. R., K. Suraj and K. K. Gandhi. 2000. Caprine sperm acrosome reaction: Promotion by progesterone and homologous zona pellucida. Small Rum. Res. 37:279-286.
  10. Walensky, L. D. and S. H. Snyder. 1995. Inositol 1,4,5-trisphosphate receptors selectively localized to the acrosomes of mammalian sperm. The J. Cell. Biol. 130:857-869.
  11. Yanagimachi, R. 1994. Mammalian fertilization. In: 'The Physiology of Reproduction' (Ed. E. Knobil and J. D. Neil). New York: Raven Press, 2nd Ed, pp. 189-317.
  12. Benoff, S. 1998. Voltage dependent calcium channels in mammalian spermatozoa. Front. Biosci. 1(3):D1220-1240.
  13. Florman, H. M., M. E. Corron, T. D-H. Kim and D. F. Babcock. 1992. Activation of voltage dependent Ca2+ channels of mammalian sperm is required for zona pellucida induced acrosomal exocytosis. Dev. Biol. 152:304-311.
  14. Kaul, G., S. Singh, K. K. Gandhi and S. R. Anand. 1997. Calcium requirement and time course of capacitation of goat spermatozoa assessed by chlortetracycline assay. Andrologia 24:243-251.
  15. Revelli, A., M. Modotti, A. Pittareth-Yanez, M. Massobrio and M. Balerna. 1994. Steroid receptors in human spermatozoa. Hum. Reprod. 2:317-326.
  16. Fraser, L. R. and K. McIntyre. 1989. Calcium channel antagonists modulate the acrosome reaction but not capacitation in mouse spermatozoa. J. Reprod. Fertil. 86:223-233.
  17. Murphy, S. J., E. R. S. Roldan and R. Yanagimachi. 1986. Effects of extracellular cations and energy substances on the acrosome reaction of precapacitated guinea pig spermatozoa. Gamete Res. 14:1-10.
  18. Publicover, S. J. and C. L. R. Barrat. 1999. Voltage operated $Ca^{2+}$ channels and the acrosome reaction : which channels are present and what do they do? Hum. Reprod. 14:873-879.
  19. Schuetz, A. W. and N. H. Dubin. 1981. Progesterone and prostaglandin secretion by ovulated rat cumulus cell-oocyte complexes. Endocrinology 108:457-463.
  20. Blackmore, P. F. and S. Eisoldt. 1999. The neoglycoprotein mannose-bovine serum albumin, but not progesterone, activates T-type calcium channels in human spermatozoa. Mol. Hum. Reprod. 5(6):498-506.
  21. Cross, N. L., P. Morales, J. M. Overstreet and F. W. Hanson. 1988. Induction of acrosome reaction by human zona pellucida. Biol. Reprod. 38:235-244.
  22. Hyne, R. V. 1984. Bicarbonate- and calcium- dependent induction of rapid guinea pig sperm acrosome reactions by monovalent ionophores. Biol. Reprod. 31:312-323.
  23. O'Toole, C. M. B., E. R. S. Roldan and L. R. Fraser. 1996. Role of $Ca^{2+}$ channels in the signal transduction pathway leading to acrosomal exocytosis in human spermatozoa. Mol. Reprod. Dev. 45:204-211.<204::AID-MRD13>3.0.CO;2-0
  24. Shi, Q. X., Y. Y. Yuan, X. Y. Lia and E. R. S. Roldan. 1996. GABA initiation of acrosome reaction mammalian sperm and interaction with progesterone. Biol. Reprod. 54(Suppl 1): 60.
  25. Fraser, L. R. 1993. Calcium channels play a pivotal role in the sequence of ionic changes involved in initiation of mouse sperm acrosomal exocytosis. Mol. Reprod. Dev. 36:368-379.
  26. Blackmore, P. F. 1993. Rapid non-genomic actions of progesterone stimulate $Ca^{2+}$ influx and the acrosome reaction in human sperm. Cell. Signall. 5:531-538.
  27. Austin, C. R. 1951. Observation on the penetration of sperm into the mammalian egg. Aust. J. Sci. Res. B4:697-698.
  28. Tiwari-Woodruff, S. K. and T. C. Cox. 1995. Boar sperm plasma membrane Ca2+ selective channels in planar lipid bilayers. Am. J. Physiol. 268 (Cell. Physiol. 37):C1284-C1294.
  29. Bleil, J. D. and P. M. Wassarman. 1983. Sperm-egg interactions in the mouse: Sequence of events and induction of acrosome reaction by a zona pellucida glycoprotein. Dev. Biol. 95:317-324.
  30. Hyne, R.V., R. E. Higginson, D. Kohlman and A. Lopata. 1984. Sodium requirement for capacitation and membrane fusion during the guinea-pig sperm acrosome reaction. J. Reprod. Fertil. 70:83-94.
  31. Meyers, S. A., J. W. Overstreet, I. K. M. Liu and E. Z. Drobnis. 1995. Capacitation in vitro of stallion spermatozoa: Comparison of progesterone induced acrosome reactions in fertile and subfertile males. J. Androl. 16:47-54.
  32. Shi, Q. X., Y. Y. Yuan and Z. H. Zhang. 1992. Factor(s) affecting the acrosome reaction in Chinese hamster sperm. Biol. Reprod. 46(Suppl 1):9.
  33. Snedecor, G. W. and W. G. Cochran. 1967. Statistical methods. Oxford & IBH Publishing Co., Calcutta.
  34. Chang, M. C. 1951. Fertilizing capacity of spermatozoa deposited in the fallopian tubes. Nature 168:697-689.
  35. Nilius, B., P. Hess, J. B. Lansman and R. W. Tsien. 1985. A novel type of cardiac $Ca^{2+}$ channel in ventricular cells. Nature 316:433-446.
  36. Fraser, L. R., G. Umar and S. Sayed. 1993. $Na^+$-requiring mechanisms modulate capacitation and acrosomal exocytosis in mouse spermatozoa. J. Reprod. Fertil. 98:187-194.
  37. Fraser, L. R. 1990. Sperm capacitation and its modulation. In:'Fertilization in mammals' (Ed. B. D. Bavister, J. M. Cummins and E. R. S. Roldan). Serono Symposia, Norwell, MA. pp. 141-153.
  38. Melendrez, C., S. Meizel and T. Berger. 1994. Comparison of the ability of progesterone and heat solubilized porcine zona pellucida to initiate the porcine sperm acrosome reaction in vitro. Mol. Reprod. Dev. 39:433-438.
  39. Malgaroli, A., D. Milani, J. Meldolezi and T. Possan. 1987. Fura-2 measurement of cytosolic free $Ca^{2+}$ in monolayers and suspensions of various types of animal cells. The J. Cell. Biol. 105:2145-2155.
  40. Roldan, E. R. S., T. Murase and Q. X. Shi. 1994. Exocytosis in spermatozoa in response to progesterone and zona pellucida. Science 266:1578-1581.
  41. Shi, Y. L. and X. H. Ma. 1998. Ion channels reconstituted into lipid bilayer from human sperm plasma membrane. Mol. Reprod. Dev. 50:354-360.<354::AID-MRD12>3.0.CO;2-4

Cited by

  1. Calcium channels in chicken sperm regulate motility and the acrosome reaction vol.283, pp.10, 2016,


Supported by : Council of Scientific and Industrial Research