Abstract: bjective To explore the role of dopamine transporter (DAT) regulated by glial cell line-derived neurotrophic factor (GDNF) of midbrain in Parkinson’s disease (PD) as well as its mechanism. Method Adeno-associated virus were used to interfere with the ex＜x＞pression of GDNF and DAT in the midbrain of PD model mice respectively. The motor function of mice were evaluated by behavior tests. The ex＜x＞pression of GDNF, DAT and TH were detected by Western blot. The distribution of TH+ neurons in the midbrain was observed by immunofluorescence. DA levels in the midbrain and striatum were detected by ELISA. Results Down-regulation of GDNF in the midbrain of PD model mice resulted in shortened duration on the rod and total distance of activity of mice, decreased distribution of TH+ neurons and ex＜x＞pression of DAT in the midbrain, and decreased levels of DA in the midbrain and striatum. Down-regulation of DAT in the midbrain of PD model mice resulted in shortened duration on the rod and total distance of activity of mice, decreased distribution of TH+ neuronsin the midbrain, decreased ex＜x＞pression of DAT and TH in the striatum, and decreased levels of DA in the midbrain and striatum. Conclusion GDNF can regulate DA transmission in the nigrostriatal pathway through DAT.?Decreased GDNF in the midbrain may contribute to DA dysfunction through down-regulating DAT, thereby promoting the loss of DA neurons and the development of motor symptoms of PD.