To clarify changes in the muscle fibre properties of the temporal muscle related to the start of masticatory movement, we immunohistochemically investigated myosin heavy chain (MyHC) isoform protein expression using pre-weaning and post-weaning mice. In addition, we examined the expression of a gene coding for those MyHC proteins. Immediately after weaning, isoforms with fast and potent contractility were frequent. This suggests that the temporal muscle plays an important role in a marked functional change in the oral cavity from lactation to mastication, contributing to oral function in cooperation with other masticatory muscles.