Pharmacological investigation of voltage-dependent Ca²⁺ channels in human ejaculatory sperm in vitro

Abstract

The types of the voltage-dependent calcium channels (VDCCs) in human ejaculatory sperm and the effects of calcium channel blocker (CCB) on human sperm motility parameters in vitro were investigated. The human sperm motility parameters in vitro in response to the pharmacological agents nifedipine (NIF, inhibitor of L-type VDCC) and θ-contoxin (GVIA, inhibitor of N-type VDCC) were compared and analyzed statistically. The results showed that NIF (1, 5, 10 μmol/L) could not only significantly affect human sperm’s shape but also spermatozoa motility after incubated at least 10 min in vitro (P<0.001). GVIA (0.1, 0.5 and 1 μmol/L) could just only significantly affect human sperm’s progressive motility (a %+b %) after incubated for 20 min in vitro (P<0.01), but they both could not significantly affect spermic abnormality rate. It is suggested that L-type VDCC, non L-type VDCCs and isoform of L-type VDCC exist in the cell membrane of human sperm solely or together, and they participate in the spermic physiological processes especially the spermic motility.

Keywords

voltage-dependent calcium channels / calcium channel blocker / human sperm motility / in vitro

References

[1] BreitbartH.. Intracellular calcium regulation in sperm capacitation and acrosomal reaction. Mol Cell Endocrinol, 2002, 187(1–2): 139-144

[2] AllamaneniS. S., AgarwalA., RamaS., et al.. Comparative study on density gradients and swim-up preparation techniques utilizing neat and cryopreserved spermatozoa. Asian Androl, 2005, 7(1): 86-92

[3] HauserR., Godfrey-BaileyL., ChenZ.. Does the potential for selection bias in semen quality studies depend on study design? Experience from a study conducted within an infertility clinic. Hum Reprod, 2005, 20(9): 2579-2583

[4] Kirkman-BrownJ. C., BarrattC. L., PublicoverS. J.. Nifedipine reveals the existence of two discrete components of the progesterone-induced [Ca²⁺]i transient in human spermatozoa. Dev Biol, 2003, 259(1): 71-82

[5] ArikkathJ., CampbellK. P.. Auxiliary subunits: essential components of the voltage-gated calcium channel complex. Curr Opin Neurobiol, 2003, 13(3): 298-307

[6] KangM. G., CampbellK. P.. Gamma subunit of voltage-activated calcium channels. Biol Chem, 2003, 13(24): 21 315-21 318

[7] CatterallW. A.. Structure and regulation of voltage-gated Ca²⁺ channels. Annu Rev Cell Dev Biol, 2000, 16: 521-555

[8] CatterallW. A., StriessnigJ., SnutchT. P., et al.. Compendium of voltage-gated ion channels: calcium channels. Pharmacol Rev, 2003, 55(4): 579-581

[9] KoshimizuT. A., Van GoorF., TomicM., et al.. Characterization of calcium signaling by purinergic receptor-channels expressed in excitable cells. Mol Pharmacol, 2000, 58(5): 936-981

[10] DarszonA., WoodC. D., BeltranC., et al.. Measuring ion fluxes in sperm. Methods Cell Biol, 2004, 74: 545-576

[11] ShiY. L., MaX. H.. Ion-channels reconstituted into lipid bilayer from human sperm plasma membrane. Mol Reprod Dev, 1998, 50(3): 354-360

[12] Gonzalez-MartinezM. T., Bonilla-HemandezM. A., Guzman-GrenfellA. M.. Stimulation of voltage-dependent calcium channels during capacitation and by progesterone in human sperm. Arch Biochem Biophys, 2002, 408(2): 205-210

[13] BainsR., MilesD. M., CarsonR. J., AdegheJ.. Hyaluronic acid increases motility/intracellular Ca²⁺ concentration in human sperm in vitro. Arch Androl, 2001, 47(2): 119-125

[14] BlackmoreP. F., EisoldtS.. The neoglycoprotein mannose-bovine serum albumin, but not progesterone, activates T-type calcium channels in human spermatozoa. Mol Hum Reprod, 1999, 5(6): 498-506

[15] SonW. Y., LeeJ.H., HanC. T.. Acrosome reaction of human spermatozoa is mainly mediated by alpha1H T-type calcium channels. Mol Hum Reprod, 2000, 6(10): 893-897

[16] JagannathanS., PuntE. L., GuY., et al.. Identification and localization of T-type voltage-operated calcium channel subunits in human male germ cells. Expression of multiple isoforms. Biol Chem, 2002, 277(10): 8449-8456

[17] GoodwinL. O., KarabinusD. S., PergolizziR. G., et al.. L-type voltage-dependent calcium channel alpha-1C subunit mRNA is present in ejaculated human spermatozoa. Mol Hum Reprod, 2000, 6(2): 127-136

[18] ParkJ. Y., AhnH. J., GuJ. G., et al.. Molecular identification of Ca²⁺ channels in human sperm. Exp Mol Med, 2003, 35(4): 285-292

[19] TrevinoC. L., FelixR., CastellanoL. E., et al.. Expression and differential cell distribution of low-threshold Ca²⁺ channels in mammalian male germ cells and sperm. FEBS Lett, 2004, 563(1–3): 87-92