Objective To predict the target genes of Mmu-miR-124 by bioinformatics method, and to identify the role of Mmu-miR-124 in neurodevelopment. Methods The potential target genes of Mmu-miR-124 were predicted on Star Base database. GO, Reactome Pathway, KEGG Pathway and PPI analysis methods were used to analyze the possible mechanism of the predicted target genes. The reliability of the predicted results on key genes in neural developmental pathways was confirmed by real-time fluorescence quantitative reverse transcriptional PCR (qRT-PCR) technique. Results GO, KEGG and Reactome pathway suggested the role of Mmu-miR-124 target genes in axon guidance, long-term depression and sphlein metabolic pathway. PPI analysis showed that GSK3B, NRAS and ITGB1 were key genes. qRT-PCR results showed that Mmu-miR-124 was increased significantly and GSK3B was decreased slightly during the neurodevelopment. Conclusion Mmu-miR-124 is widely involved in a variety of biological processes by regulating target genes, and Mmu-miR-124 may regulate neural development by targeting GSK3B.