Abstract: The reproductive behaviors of birds are mainly controlled by the hypothalamus-pituitary-gonad axis. Many studies have shown that reproductive hormones are tightly linked to the breeding sub-stages. However, only a few studies have examined the temporal trend of hormone levels among different reproductive stages in multiple brooded species. We investigated the changes in plasma luteinizing hormone (LH) and prolactin (PRL) concentrations during different reproductive stages of the facultative double-brooded Great Tit (Parus major). We found that the concentrations of LH and PRL in females were significantly higher than those in males. Females had significantly higher LH and lower PRL concentrations in the pre-breeding period than in the first/second brooding periods, and there were no significant changes between the first and second brooding periods. The concentrations of LH and PRL in males had no significant difference between the pre-breeding period and the first brooding periods, while LH and PRL concentrations in the second brooding period were significantly higher than those in the first brooding period. We conclude that there are sex-based differences between LH and PRL at different stages of reproduction. The changes in LH and PRL in both males and females should be related to their physiological functions. Especially for males, individuals with higher levels of LH and PRL are more likely to maintain second clutches.