Methamphetamine (MA) is a highly addictive psychostimulant abused worldwide that causes long-term behavioral changes that are linked to changes in the brain. The third trimester and adolescence are critical periods of development as the body undergoes dramatic changes and becomes more sensitive to the effects of the drug. This study aimed to investigate the effect of MA exposure during the first 12 postnatal days of laboratory rats, corresponding to the human third trimester, on hippocampal neurotrophin levels in adolescence. Exposure was performed either directly (daily subcutaneous injections) or indirectly (by breastfeeding-exposed mothers). Brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) were analyzed in the rat hippocampus. Rats were also exposed to different housing conditions before weaning (enriched environment (EE) or standard) and after weaning (group vs. separate). Adolescence was divided into 3 age categories: postnatal day (PD) 28, which corresponds to juvenile age (early adolescence), PD 35 - adolescence, PD 45 - late adolescence. The results of this study showed that EE did not increase BDNF levels in either control or MA-exposed animals, and these results are similar for both direct and indirect exposure. On the other hand, social separation after weaning reduced BDNF levels compared to animals reared in a standard environment, but this effect was reversed by direct exposure to MA. As for NGF, EE increased its levels only indirectly exposed control animals and animals exposed to MA during late adolescence, as separation increased these levels in most animals. Funding: Research program Cooperatio Neurosciences from Charles University