Increasing evidence indicates that nerves play a significant role in regulating stem cell homeostasis and developmental processes. To explore the impact of nerves on epithelial stem cell homeostasis during tooth development, the regulation of sensory nerves on stem cell homeostasis was investigated using a rat model of incisor development. Impaired mineralization, decreased enamel thickness, and fractured enamel rods of the incisor were observed after denervation. qPCR and histological staining revealed that the expression of enamel-related factors ameloblastin (AMBN), kallikrein-4, amelogenin (Amelx), collagen type XVII (col17a), and enamelin were decreased in the incisor enamel of rats with sensory nerve injure. The decreased expression of Wnt5a in ameloblasts was coupled with the downregulation of calcium ion-related calmodulin kinase II. These results implicate that the sensory nerves are essential in stem cell homeostasis for enamel mineralization and development.