This study was aimed at determining, with homologous mouse gametes, whether a level of membrane lipid peroxidation insufficient to affect sperm motility parameters can alter sperm fertilizing potential. The addition of ferrous ions and ascorbic acid (Fe2+/ Asc) to mouse sperm suspensions increases the formation of thiobarbituric acid-reactive substances (TBARS), an indicator of lipid peroxide breakdown products, without significantly affecting the level of sulfhydryl groups. In the presence of Fe2+/Asc concentrations above > 0.4/2.0 mM, spermatozoa become immotile. However, at concentrations < or = 0.4/2.0 mM of Fe2+/Asc, i.e., conditions in which the TBARS formation is increased by < or = 4.6-fold over that of controls, motility remains unaffected for up to 3 hours. In the presence of 0.4/ 2.0 mM Fe2+/Asc, treated spermatozoa increase their fertilizing potential by 50%, as measured by the formation of two-cell embryos. This increase is not caused by improvements in sperm motility parameters (percentage, linearity, velocity, hyperactivation) or sperm capacitation. On the other hand, there is a significant increase in the capacity of mouse spermatozoa to bind to homologous zona pellucida. In conclusion, mild peroxidative conditions, that increase lipid peroxide formation 4.6-fold without significantly modifying free sulfhydryl groups and sperm motility parameters, improve the fertilizing potential of spermatozoa by increasing their binding capacity to zona pellucida.