Sensitive detection of myosin heavy chain composition in skeletal muscle under different loading conditions.

Different loading conditions were employed to study changes in myosin heavy chain (MHC) composition with the aid of a sensitive approach for separating and detecting MHC isoforms. Separation and detection of MHCs by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were achieved to a degree such that MHC composition is consistent with previous reports on functional and mRNA data. Neonatal MHC was detected at low levels in control, 14-day hindlimb unweighted (HU), and 28-day HU soleus muscles. Type IIa MHC remained unchanged in all groups, representing approximately 9% of total MHC present. Type IIb MHC was not detected in control but represented 3% of total MHC at both 14 and 28 days of HU. Type IIx MHC also was not detected in control but represented 6% of total MHC at 14 days HU, and increased to 14% of total MHC at 28 days HU (P < 0.05). Type I MHC decreased from 89% in control to 72% at 28 days HU (P < 0.05). Furthermore, the type I MHC band was separated into two bands of approximately equal content in all groups when low amounts of protein were loaded on gels. The decrease in type I MHC with HU could be attributed entirely to a decrease in the percentage of the band in the type I region with lower mobility, which corresponds to beta-MHC. In addition, hypertrophied plantaris muscles demonstrated a fast-to-slow shift in MHC composition as evidenced by increased I, IIa, and IIx MHC and decreased IIb MHC expression.