Myotonic dystrophy (DM1) is caused by the expansion of a CTG repeat in the noncoding region of a protein kinase, DMPK, expressed in skeletal and cardiac muscles. The aim of the present study was to determine the effects of very large CTG expansions on DMPK expression and skeletal muscle development. In fetuses suffering from the severe congenital form of DM1 with large CTG expansions (1800 to 3700 repeats), the skeletal muscle level of DMPK was reduced to 57% of control levels and a similar reduction was observed in cultured DM1 muscle cells relative to control cultures. These results are consistent with greatly reduced DMPK expression from the mutant allele and normal expression from the unaffected allele in this autosomal dominant disorder. In normal fetuses, DMPK protein levels increased dramatically between 9 and 16 weeks and remained high throughout the remaining gestation period. DM1 fetuses showed impaired skeletal muscle development, characterized by a persistence of embryonic and fetal myosin heavy chains and almost total absence of slow myosin heavy chains at the end of gestation. DMPK expression, however, was similar in both fast and slow fibers from normal adult muscle. The reduced DMPK and the delayed slow fiber maturation in congenital DM1 may be two separate consequences of nuclear retention of DMPK RNA transcripts with expanded CUG repeats.