The Enzymatic Antioxidant System of Human Spermatozoa

Figure 3

PRDX6 differentially reacts with different ROS in human spermatozoa. Percoll-washed spermatozoa were incubated with different concentrations of H₂O₂, ONOO⁻, or tert-buthyl hydroperoxide (tert-BHP) for 30 min at 37°C and then sperm proteins were electrophoresed and immunoblotted with anti-PRDX6 antibody as previously described [110]. PRDX6 is present as a doublet observed in absence of H₂O₂ or 0.05 mM ONOO⁻ that becomes a single band with high intensity upon increased concentrations of ROS. High levels of H₂O₂ (but not of ONOO⁻ or tert-BHP) generate the formation of high molecular mass complexes. Representative blot from other four experiments performed with different healthy donors.