Myosin heavy chain composition of different skeletal muscles in Large White and Meishan pigs

Four major sarcomeric myosin heavy chains (MyHC) (i.e., I, IIa, IIx, and IIb) are expressed in pig skeletal muscle during postnatal development. The objective of the current study was to compare MyHC composition at mRNA and protein levels in LM, a fast-twitch glycolytic muscle, and rhomboideus (RM), a mixed slow- and fast-twitch oxido-glycolytic muscle, between two pig breeds exhibiting dramatic differences in postnatal muscle growth and meat quality. Eight Large White (LW) and eight Meishan (MS) females were fed under the same standard conditions, and slaughtered at an average BW of 62 kg (131 and 142 d in LW and MS pigs, respectively). In addition to conventional fiber typing by histoenzymology, MyHC composition was analyzed by combining immunocytochemistry, in situ hybridization, and a newly developed real-time PCR assay. Enzyme activities of lactate dehydrogenase, citrate synthase, and beta-hydroxy-acyl-CoA-dehydrogenase were used as markers of glycolytic, oxidative and beta-oxidation capacities, respectively. Results showed that conventional fiber typing in three classes by histoenzymology was insufficient in LM. For the first time, four monoclonal antibodies specific of each MyHC isoform, working in immunocytochemistry, were used. Our results are consistent with the sequential I<-->IIa<-->IIx<-->IIb MyHC transition rule. Breed effect on MyHC composition differed between muscle types. In LM of MS pigs, a shift from IIb to IIx, and to a lesser extent, to IIa, occurred without affecting type I MyHC. In RM, where IIb is absent, a shift from IIx to type I occurred, with a slight decrease in the IIa isoform. Effects were very similar at the mRNA and protein levels, suggesting a transcriptional regulation. In both muscles, MS pigs exhibited a decrease in the relative fiber type specific expression of the fastest isoform (i.e., IIb in LM and IIx in RM). The shift toward a slower phenotype in MS pigs was consistent with a less glycolytic and more oxidative metabolism, potentially using more lipids as fuel. A dramatic increase in cross-sectional area of type I fibers in RM (+27%) and a decrease in that of the fastest IIb fibers in LM (-25%) were observed in MS pigs. Overall, interpretation of earlier data regarding muscle fiber type has been flawed by inaccurate fiber typing in most pig skeletal muscles.