ObjectiveTo quantitatively analyze myosin heavy chain (MHC) mRNA composition in two rat middle ear muscles (the tensor tympani and stapedius) using competitive polymerase chain reaction (PCR).Material and MethodsAn exogenous template, including oligonucleotide sequences specific for the seven rat MHCs (2A, 2B, 2X, 2L/EOM, embryonic, neonatal and β-cardiac) as well as β-actin, was constructed and used as the competitor.ResultsThe tensor tympani and stapedius contained all MHC isoforms except 2L. The tensor tympani contained approximately equal proportions of 2X (40.4%±6.5%) and 2A (34.0%±1.3%) MHCs, with a smaller percentage of 2B (16.6%±1.5%) and neonatal (7.5%±0.6%) MHCs, while β-cardiac and embryonic MHCs were minimally expressed. The stapedius contained predominantly 2X (58.0%±4.2%) and 2A (32.3%±6.7%) MHCs, with a smaller percentage of 2B (7.4%±0.2%) and β-cardiac (1.9%±0.1%) MHCs. Neonatal and embryonic MHCs were detected at very low levels.ConclusionThese results suggest that two middle ear muscles, which are mainly composed of two fast-twitching myosins (2X and 2A MHCs), contract fast and are fatigue-resistant.