The digastric muscle is one of the suprahyoid muscles and consists of the anterior and posterior bellies. Because muscle fiber alignments in these two bellies are different, the functional roles are said to be different. Since the digastric muscle relates to mastication, its functions may change markedly before and after weaning, but many details remain unknown. The aim of this study was to clarify changes in muscle fiber properties of the anterior and posterior bellies of the digastric muscle in mice before and after weaning. Expressions of myosin heavy chain (MyHC) isoforms were assessed at the protein and transcriptional levels. Expression of the MyHC-2b isoform, an isoform displaying fast, strong contraction, was greater in the anterior belly than in the posterior belly after weaning. This suggests that, in mice, the anterior belly of the digastric muscle needs to move rapidly anteroposteriorly for mastication, compared with the posterior belly.