Objective To investigate the effects of aerobic exercise on the inhibitory synaptic plasticity of the ventral tegmental area (VTA) dopamine neurons of mice with nicotine withdrawal. Methods Eight-week-old male C57BL/ 6 mice were randomly divided into a saline sedentary (SS) group, saline exercise ( SE) group, nicotine sedentary (NS) group, and nicotine exercise (NE) group following the conditioned place preference (CPP) paradigm. During abstinence, SE and NE group mice were subjected to a moderate-intensity treadmill running protocol for 2 weeks. The CPP paradigm was used to test nicotine-seeking behavior of mice after nicotine withdrawal. The whole-cell patch clamp was used to record the spontaneous inhibitory postsynaptic current ( sIPSC), the miniature IPSC (mIPSC), and the IPSC paired-pulse ratio (PPR) of dopamine neurons in the VTA. Immunofluorescence staining was used to detect the expression of GAD67 in the VTA. Results (1) The CPP score was higher in the NS than SS group and lower in the NE than NS group. (2) The mean frequency and amplitude of the sIPSC were higher in the NS than SS group and lower in the NE than NS group. (3) The mean frequency and amplitude of the mIPSC were higher in the NS than SS group and lower in the NE than NS group. (4) At the 20 ms and 50 ms interstimulus intervals, the IPSC PPR was lower in the NS than SS group and higher in the NE than NS group. (5) The fluorescence intensity of GAD67 was lower in the NS than SS group and higher in the NE than NS group. Conclusions 2 weeks moderate intensity aerobic exercise reduces presynaptic GABA release, down-regulates the number and / or function of postsynaptic GABAA receptors in VTA dopamine neurons, and these decreased inhibitory synaptic transmission may represent a potential mechanism by which aerobic exercise attenuates nicotine seeking during abstinence.