Functional maturation of muscles is related to the constitutional proportion of muscle protein isoforms during development and growth. Although the mouse masseter muscle (MS) is classified as a fast limb muscle, its functions are different from those of a limb muscle. This study investigated the differentiation of myosin heavy chain (MHC) isoforms during the postnatal development periods in mouse MS and mouse tibialis anterior (TA), which is a fast limb muscle. Many anti-MHC slow-type-positive fibres were observed in neonatal MS and TA; these fibres decreased during development. Adult MS was composed of anti-MHC fast-type-positive fibres. MHC isoforms in MS were composed of MHC-2a and MHC-2d soon after birth. MHC-2b was expressed, but MHC-2a was not seen after 21 days. Expression of MHC-2b agreed with the weaning period, that is 2-3 weeks after birth. This fact suggested that the transformation from suckling to mastication changed the MHC isoforms during this period. In this study, the expressions of MHC-2b agree with the weaning period.