Effects of chronic stimulation with different impulse patterns on the expression of myosin isoforms in rat myotube cultures.

In order to study maturation and differentiation of aneural myotubes in vitro, long-term myotube cultures were established from hindlimb musculature of newborn rats. The developmental state of the myotubes was judged by their myosin heavy chain (HC) patterns. Newly formed myotubes only expressed the embryonic isoform, HCemb, older myotubes expressed the neonatal isoform HCneo, as well as the fast adult isoforms HCIIb and HCIId. HCIId increased continuously, reaching a relative concentration of 47% in 37-day-old cultures. The third fast isoform, HCIIa, was not detected and also the slow isoform HCI was absent. Effects of chronic (20 days) electrostimulation were studied by exposing the cultures to various stimulus patterns. Bursts of 250 ms duration at various pulse frequencies were applied at low and high burst frequencies. Although HCemb remained the predominant isoform under all conditions, different stimulus patterns induced specific changes in the patterns of fast and slow HC isoforms. Bursts of 250 ms duration at 15 Hz, 40 Hz, or 100 Hz, repeated every second or every 4 s, induced the expression of slow myosin, i.e., HCl. Bursts of 250 ms duration at 100 Hz, repeated every 100 s, enhanced the expression of HCIId, but not of HCI. Because slow myosin was induced at high burst frequency with low and high pulse rates, we suggest that burst frequency rather than pulse frequency has a specifying effect on myosin expression. Our results show that the basal program of myosin expression during myogenesis in vitro can be modulated by electrostimulation, suggesting a possible influence of neuromuscular activity on the development of adult fiber types.