Reactive oxygen species (ROS) are well-established modulators of luteal cell apoptosis in the estrous cycle. The objective of this study was to clarify the molecular mechanisms of luteolysis by characterizing the levels and regions of mRNAs involved in ROS-induced luteal cell apoptosis. Stimulation of bovine luteal cells by H2O2 resulted in the induction of apoptotic nuclear condensation and Caspase-3 activation. In addition, a marker for oxidative stress-damaged DNA, 8-hydroxy-2' -deoxyguanosine, was highly accumulated in the large luteal cells prepared from the late estrous stage. Reverse transcription polymerase chain reaction and Northern blot analysis demonstrated that mRNAs of cyclooxygenase (COX)-2, p53, and Bax were highly accumulated in the H2O2-treated cells. In situ hybridization revealed that these mRNAs were most abundantly expressed in the large luteal cells. These findings suggest that enhancement of ROS in the bovine corpus luteum induces expression of COX-2, p53, and Bax mRNAs, resulting in activation of the signaling pathway for luteal-cell apoptosis.