The chronic administration of nafenopin, a hypolipidemic drug, induced an increase in catalase and acyl-CoA oxidase activities in various skeletal muscles, including the gracilis, diaphragm, soleus, and extensor digitorum longus. The magnitude of the increase was around 100% for both enzymes in each of the muscles studied in spite of the different basal level. These changes seem to be specific of the peroxisomal enzymes because acetylcholinesterase, which is not peroxisomal, did not follow the same pattern in all the muscles. Concomitant with the increase in muscle peroxisomal enzymes, the skeletal muscles presented an altered electromyogram with prolonged insertional activity, repetitive firing of action potentials, and myotonic runs characteristic of myotonia. Our results suggest a role for peroxisomes in the myotonic disorder.