Drosophila were examined to see if there is an increase in damage associated with free radical activity in older flies. The levels of superoxide radical and of lipid peroxides were higher in membrane samples from older flies. Also, in older Drosophila there was a significant decrease in membrane fluidity, as determined by fluorescence polarization, and an increase in ATP-dependant calcium uptake. In homogenates from whole flies, the concentration of inorganic peroxides and activity of the proteolytic enzyme, cathepsin B, increased with age. During their lifetime, vestigial wing Drosophila displayed a greater level of free radical activity than wildtype flies and a significantly shorter life span (26.7 +/- 0.7 days vs. 34.4 +/- 1.0, p < 0.01). These results indicate that the level of oxidative stress is closely coupled to cellular damage and to life span, and they indicate that free radicals may play a central role in the aging process in Drosophila.