We show that members of the POU homeodomain family are among the transcription factors expressed in developing mouse skeletal muscle. From a cDNA library prepared from fetal muscle mRNA, we cloned a cDNA identical to that of Brn-4, a POU class II gene previously cloned from neural tissues. In limb muscle, we found that Brn-4 mRNA expression was highest at embryonic days 15-18, declined-after birth, and was undetectable in adults. The mRNAs of two additional POU genes, Emb (POU class VI) and Oct-1 (POU class II), were also expressed in developing muscle and, unlike Brn-4, continued to be expressed in postnatal and adult muscles. In skeletal muscle, expression of Brn-4 is myogenin-dependent, because muscles from myogenin-deficient fetuses contained much less Brn-4 mRNA than muscles from myogenin-expressing littermates. In contrast, expression of Emb was the same in the presence or absence of myogenin. The distinct pattern of Brn-4 mRNA expression and its dependence on a myogenic regulatory factor suggest that Brn-4 is part of the network of interacting transcription factors that control muscle-specific gene expression during mammalian myogenesis.