Adult fast and slow skeletal muscles are composed of a large number of fibers with different physiological and biochemical properties that under neuronal control can respond in a plastic manner to a variety of stimuli. Although muscle cells synthesize muscle-specific contractile proteins in the absence of motoneurons, after innervation the neuron controls the particular set of isoforms subsequently synthesized. However, agreement has not been reached on the mechanism, either chemotrophic or impulse-mediated, by which the nerve influences gene expression in the muscle. Here we report the effect on isomyosins of continuous, low-frequency (a protocol mimicking the discharge pattern of the slow motoneuron) direct electrical stimulation of a permanently denervated fast muscle, the extensor digitorum longus of adult rat. After several weeks, unlike sham-stimulated muscle, the stimulated muscle showed a dramatic increase of the slow myosin light and heavy chains. Myosin light chains were identified by two-dimensional gel electrophoresis. The slow myosin heavy chain was clearly distinguished from fast and embryonic types by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and orthogonal peptide mapping. The myosin change could be restricted to a portion of the muscle by the position of the stimulating electrodes. Taking into account the morphologic appearance of the electrostimulated muscle and the large body of evidence demonstrating the absolute dependence of slow myosin on specific innervation, our observations indicate that at least the slow motoneuron influences the isomyosin genes' expression by the kind of activity it imposes on developing muscle fibers.