Molecular Human Reproduction, Vol. 9, No. 11, 645-661,
November 2003
© 2003 European Society of Human Reproduction and Embryology
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Multiple forms of redox activity in populations of human spermatozoa
Submitted on June 23, 2003; accepted on July 14, 2003
ARC Centre of Excellence in Biotechnology and Development and Discipline of Biological Sciences, School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, NSW 2308, Australia
1 To whom correspondence should be addressed at: Discipline of Biological Sciences, School of Environmental and Life Sciences, University of Newcastle, University Drive, Callaghan, NSW 2308, Australia. e-mail: jaitken{at}mail.newcastle.edu.au
In this study we have examined the biochemical attributes of the redox systems that regulate human sperm function using 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulphophenyl)-2H-tetrazolium, monosodium salt (WST-1), lucigenin and luminol-peroxidase as probes. WST-1 was readily reduced by human spermatozoa in the presence of an intermediate electron acceptor (IEA) or NAD(P)H. The IEA-mediated activity resembled a previously described trans-membrane NADH oxidase in being inhibited by capsaicin, superoxide dismutase (SOD) and N-ethyl maleimide, but differed in its sensitivity to p-chloromercuriphenylsulphonic acid (pCMBS). The NAD(P)H-induced WST-1 reduction resembled the superficial oxidase described previously, in its sensitivity to pCMBS, but differed in its suppression by capsaicin. Lucigenin was reduced by human spermatozoa in a manner that could be inhibited by SOD and stimulated by NAD(P)H or 12-myristate, 13-acetate phorbol ester. A23187 also stimulated human spermatozoa via a diphenylene iodonium-sensitive pathway detectable with luminol-peroxidase but not lucigenin. Defective sperm populations recovered from the low-density region of Percoll gradients were characterized by high levels of redox activity that was only discernable with lucigenin. We conclude that human spermatozoa possess multiple plasma membrane redox systems that are involved to varying extents in the physiological control and pathological disruption of sperm function. Their distinct pharmacological profiles should significantly assist attempts to resolve and characterize these systems.
Key words: human spermatozoa/lucigenin/luminol/Percoll fractionation/redox activity
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