In rats, mating at postpartum estrus and delayed dispersal of the young would result in the overlapping of two different-age litters. As a consequence, newborn pups' early experience will include not only that acquired during the interaction with the mother and age-matched littermates, but also with older siblings. As early-life experience modulates rodents' brain function, behavior and reproduction, we aimed to assess how changes in the early environment provoked by the overlapping of litters would affect emotionality, stress response and reproductive functions of male and female pups during adulthood. Results showed that both male and female overlapped reared pups exhibited a reduced behavioral inhibition in the open field test during adulthood. In addition, overlapped reared adult females, but not males, showed a blunted corticosterone response to an acute stressor during diestrus and a reduction in sexual behavior. In summary, natural changes in early experience provoked by the overlapping of litters, long-term modulate affective and reproductive behaviors, and the endocrine stress response in a sex dimorphic manner.