The effect of reactive oxygen species on contractions in beta-escin permeabilized rat detrusor was investigated. Cumulative calcium contractions were inhibited by hydrogen peroxide (H(2)O(2)) and hydroxyl (*OH) but not by superoxide (O(2) *). The sarcoplasmic reticulum calcium-ATPase inhibitor cyclopiazonic acid (CPA) and the mitochondrial blocker carbonyl cyanide p-trifluromethoxyphenylhydrazone (FCCP) decreased the calcium contractions, however in their presence, H(2)O(2) and *OH did not have further effect. Carbachol contractions were inhibited by either H(2)O(2)/*OH/O(2) * or CPA/FCCP. In the presence of CPA, carbachol contractions were not affected by H(2)O(2) and *OH but further decreased by O(2) *. On the other hand, only H(2)O(2) and *OH elicited additional inhibition in carbachol responses in the presence of FCCP. Inositol triphosphate contraction was inhibited by *OH whereas none of the radicals affect carbachol induced calcium sensitization. These results show that H(2)O(2) and *OH affects sarcoplasmic reticulum where O(2) * acts on mitochondria to change contractions in rat detrusor smooth muscle.