Objective：To investigate the proteomics of primary neurons exposed to methamphetamine(METH)and the corresponding cellular functions，unraveling the potential neurotoxicity of METH exposure. Methods：The primary cultured neurons were treated with METH(0，900 μmol/L)and then hydrolyzed by protease. LC⁃MS/MS was used to detect and quantify the proteins， then the database of Uniprot RattusNorvegicus_ 36080_ 20180123 was used to identify the target protein，gene ontology(GO)was used to annotate the protein function，and Kyoto Encyclopedia of Genes and Genomes(KEGG)database was used to classify the target protein sequence by KO analysis to obtain the target protein associated signal pathways. Based on the above pathways，Western blot was used to verify the synaptic damage induced by METH，and immunofluorescence was performed to examine the expression and spatial localization of synapses. Results：The total number of proteins identified was 6619. Compared with the control group，the protein with expression difference more than 1.2 times and P ＜ 0.05 was classified as differential protein. Through screening，51 different proteins were screened，of which 40 proteins were up ⁃ regulated while 11 proteins were down ⁃ regulated. For protein expression，METH exposure significantly decreased the expression of post synaptic density protein 95(PSD95)and Drebrin，while presynaptic marker synapsin 1 was significantly up ⁃ regulated；immunofluorescence results showed that Drebrin expression was significantly decreased accompanied by reduced co ⁃ localization with F ⁃ actin，meanwhile，the spatial localization of PSD95 and synapsin 1 was significantly decreased. Conclusion：METH exposure causes abnormal expression of numerous proteins in primary neurons as well as the disorderof a spectrum of cellular functions，which finalizes the destruction of synaptic structure and neuronal damage.