Cannabidiol (CBD) is a non-psychoactive substance derived from marijuana. Although a comprehensive understanding of CBD's mechanism of action is still lacking, it is well-established that CBD can effectively mitigate the addictive properties associated with drugs. This study examined how CBD inhibits the acquisition and expression of methamphetamine-induced conditioned place preference (CPP) through D2-like dopamine receptors (D2R) in the ventral tegmental area (VTA). After recovery from surgery, animals were subjected to bilateral intra-VTA administration of different dosages (0.25, 1, and 4 μg/0.3 μl DMSO per side) of a selective D2R antagonist, sulpiride before intracerebroventricular (ICV) injection of CBD, during the conditioning phase (10 μg/5 μl DMSO) or once in the post-conditioning phase (50 μg/5 μl DMSO) of methamphetamine-induced CPP (daily subcutaneous injections of methamphetamine at 1 mg/kg over 5-day conditioning period). Findings revealed that CBD inhibits the acquisition and expression of methamphetamine reward memory. At the same time, microinjection of D2R antagonists into the VTA significantly reduced CBD's suppressive effect on the acquisition (0.25 μg
P<
0.05, 1 and 4 μg
P<
0.001) and expression (1 and 4 μg
P<
0.01) of methamphetamine place preference. Moreover, D2R blockage alone in the VTA did not affect the formation and expression of methamphetamine-induced CPP. In addition, the present study showed that administration of intra-VTA sulpiride and ICV injection of CBD together does not cause place preference in the CPP paradigm. In conclusion, pharmacological manipulation of D2Rs in the VTA may alter CBD's suppressive effects on the methamphetamine-context associations.