Open Access Research

Molecular and functional characterization of voltage-gated sodium channels in human sperm

Francisco M Pinto1, Cristina G Ravina2, Manuel Fernández-Sánchez2, Manuel Gallardo-Castro1, Antonio Cejudo-Román1 and Luz Candenas1*

Author Affiliations

1 Instituto de Investigaciones Químicas, CSIC, Avda. Americo Vespucio 49, 41092 Sevilla, Spain

2 IVI-Sevilla, Avenida Republica Argentina 58, 41011 Sevilla, Spain

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Reproductive Biology and Endocrinology 2009, 7:71  doi:10.1186/1477-7827-7-71

Published: 16 July 2009

Abstract

Background

We have investigated the expression of voltage-gated sodium channels in human spermatozoa and characterized their role in sperm motility.

Methods

Freshly ejaculated semen was collected from thirty normozoospermic human donors, with each donor supplying 2 different samples. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence techniques were used to detect the mRNAs and proteins of interest. Sperm motility was measured by a computer-assisted sperm analysis system (CASA). Cytosolic free calcium was determined by fluorimetry in cells loaded with the fluorescent calcium indicator Fura-2.

Results

The mRNAs that encode the different Nav alpha subunits (Nav1.1-1.9) were all expressed in capacitated human spermatozoa. The mRNAs of the auxiliary subunits beta1, beta3 and beta4 were also present. Immunofluorescence studies showed that, with the exception of Nav1.1 and Nav1.3, the Nav channel proteins were present in sperm cells and show specific and different sites of localization. Veratridine, a voltage-gated sodium channel activator, caused time- and concentration-dependent increases in progressive sperm motility. In sperm suspensions loaded with Fura-2, veratridine did not modify intracellular free calcium levels.

Conclusion

This research shows the presence of voltage-gated sodium channels in human sperm and supports a role for these channels in the regulation of mature sperm function.