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Maternal pre-pregnancy nicotine exposure could lead to depression-like behaviors in the offspring.Biparental pre-pregnancy nicotine exposure mainly caused a depressive phenotype among F1 mice, and a few with hyperactivity.The impact of maternal nicotine exposure on the behavior of the offspring was far severer than that of paternal exposure.These effects appear to be mediated via disruption of the balance between GSK3 and p-GKS3 by nicotine.Our previous studies showed that paternal nicotine exposure can lead to hyperactivity in the offspring. Nevertheless, the cross-generational effects of maternal and biparental nicotine exposure remain unclear. In this study, female and male mice were exposed respectively by nicotine before pregnancy. The maternal pre-pregnancy nicotine exposure led to depression-like behaviors in the F1 offspring. However, after biparental pre-pregnancy nicotine exposure, seventy percentage of the offspring exhibited a depressive phenotype while 20% were hyperactive, and the remaining exhibited no obvious abnormal behavior. The cross-generational effects appeared to be mediated via disruption of the balance between GSK3 and p-GKS3 by nicotine. These results suggested that pre-pregnancy nicotine exposure can induce alterations in the behavior of the offspring, and the cross-generational effects of maternal nicotine exposure were particularly serious.