Objective:To investigate whether intracellular zinc depletion can actually change spermatozoa motility and the abundance of voltage-dependent anion channel(VDAC) mRNA in cultured human spermatozoa. Methods: Spermatozoa were separated from ejaculates of three donors using a discontinuous Percoll gradient centrifugation. Each spermatozoa specimen were divided into 3 groups,one was exposed to a cell membrane-permeable zinc chelator N,N,N＇,N＇-tetrakis(2-pyridylmethy1) Ethylenediamine (TPEN) (2 -滋mol/L) for 24 hours,one was to TPEN plus zinc sulfate(5 -滋mol/L) for 24 hours,and one was as control. CASA and quantitative real-time PCR(qPCR) were performed to detect sperm motility characteristics,mRNA abundance and difference of the three VDAC subtypes between three different treatment groups respectively. Results:The results of CASA demonstrated that exposure of spermatozoa to TPEN for three hours significantly decreased sperm motility compared with the control group(P = 0.049). qPCR demonstrated that VDAC3 mRNA level of TPEN group were significantly lower than that of control group,while no significant difference was found between TPEN plus zinc sulfate treatment group and control group. Co-addition of zinc partly reversed TPEN-induced alterations of sperm motility and VDAC3 mRNA abundance. Conclusion:The results of the present study implicate that zinc depletion may induce VDAC3 mRNA abundance decreasing and affect the normal function of spermatozoa.