We examined the influence of early weaning on the development of play-fighting behaviors and anxiety status in Wistar rats. Pups were divided into two groups, those weaned at postnatal day (PD) 16 (early-weaned group) and those weaned at PD30 (normally weaned group), and were housed in pairs of the same sex. Playful interactions were measured for each pair once a week from 4 to 7 weeks of age. Thereafter, during early adulthood, all the rats were subjected to the elevated plus-maze test. The frequencies of pinning and playful attack were less in the early-weaned group than in the normally weaned group. In the elevated plus-maze test, rat pups in the early-weaned group had higher anxiety levels. The results showed that deprivation of mother-pup interactions during the preweaning period decreases affiliative interactions between cage mates, including play-fighting behaviors during the postweaning developmental period, and increases anxiety levels during early adulthood.