Abstract： Objective To explore the mechanism of opioid-psychic dependence involving the aspects of pre-receptor and receptor by observing the changeable expressions of glutamate neurotransmitter and NR2B of N-methyl-D-aspartic acid (NMDA) receptor in the neuroanatomical circuit of ventral temental area, nucleus accumbens and prefrontal cortex (VTA-Nac-PFC) of rats subjected to morphine-induced conditioned place preference (CPP). Methods The models of CPP were validated by escalating doses of morphine in rats (n=16). The colorimetry and immunohistochemistry ways were applied to detect the contents of glutamic acid and the expression level of NR2B in VTA, Nac and PFC. Results As compared with those in the control group physiological saline), the prolonged detention time of white compartment in the model group was notably observed (P<0.05), and increased content of glutamic acid and expression level of NR2B in fields of VTA, Nac and PFC in the model group were significantly detected (P<0.05). Conclusion Increased level of giutamic acid and expression level of NR2B in nuroanatomieal circuit of VTA, Nac and PFC could play key roles in inducing morphine-psychic dependent rats.