Objective To observe the effect of overexpression of lysosome associated membrane protein 2A (LAMP2A) up-regulating chaperone-mediated autophagy (CMA) in the substantia nigra on the locomotor behavior of a Parkinson’s disease (PD) rhesus monkey model. Methods We established a rhesus monkey model of PD. A stereotactic injection device was used to inject Lewy bodies (LBs) extracts from the brain tissues donated by PD patients into the right striatum of seven rhesus monkeys. At the same time, extracts of brain tissue donated by control ( non-PD) patients were injected into the striatum of the left brain. The animals were randomly divided into two groups. The LAMP2A overexpression group (n= 4) was bilaterally injected with an adeno-associated virus vector overexpressing LAMP2A; the control group (n= 3) was injected with a control virus vector in both sides of the substantia nigra. The motor function of monkeys was evaluated by the hill and valley staircase experiment and the pick-up test. The levels of total α-synuclein ( α-syn) and phosphorylated α-syn in the cerebrospinal fluid and serum of experimental monkeys were determined by ELISA. Results In the valley staircase task and the outside board pick up test, the fine motor level of the left upper limb of the control group of rhesus monkeys was lower than that of the right side. There was no significant difference in the fine motor level of the upper limbs on the left and right sides of the rhesus monkeys in the LAMP2A overexpression group. Cerebrospinal fluid and serum total α-syn levels and phosphorylated α-syn levels of the control group increased significantly at 12 months after operation. The levels of phosphorylated α-syn in cerebrospinal fluid and serum phosphorylated α-syn in the LAMP2A overexpression group were lower than those in the control group at the same time after 8 months. Conclusions Overexpression of LAMP2A improved the fine motor ability of experimental monkeys in the rhesus monkey model of PD through up-regulating CMA-mediated α-syn elimination.