Abstract Objective To observe the impact of different dosages of rotenone on behavior and dopamine content in stratium of rats, and explore the optimum experimental condition of PD rat model induced by rotenone injection. Methods The Lewis rats were subcutaneously injected by different dosage of rotenone (1.0, 1.5 and 2.0 mg/kg/day) for 28 days. The open field and inclined plane tests were used to evaluate the movement function and coordination, respectively. High performance liquid chromatography-electrochemical method was used to measure the content of dopamine in striatum. Results The survival rate of model rats decreased with the increase of rotenone dosage. The body weight of all 3 model groups declined significantly in a dose-dependent manner. The number of lattice and the stand-up times in the open field test decreased obviously in 3 groups of rotenone rats. The maximum angle of plane in the inclined plane test declined significantly in rotenone 1.5 mg/kg group compared with control. The dopamine content in striatum of rats reduced markedly in rotenone 1.5 mg/kg group. Conclusion Chronic exposure of rotenone at the dosage of 1.5 mg/kg could reproduce the behavioral and neurochemical features of PD.