title: Expression and identification of 10 sarcomeric MyHC isoforms in human skeletal muscles of different embryological origin. Diversity and similarity in mammalian species.
abstract: In the mammalian genome, among myosin heavy chain (MyHC) isoforms a family can be identified as sarcomeric based on their molecular structure which allows thick filament formation. In this study we aimed to assess the expression of the 10 sarcomeric isoforms in human skeletal muscles, adopting this species as a reference for comparison with all other mammalian species. To this aim, we set up the condition for quantitative Real Time PCR assay to detect and quantify MyHC mRNA expression in a wide variety of human muscles from somitic, presomitic and preotic origin. Specific patterns of expression of the following genes MYH1, MYH2, MYH3, MYH4, MYH6, MYH7, MYH8, MYH13, MYH14/7b and MYH15 were demonstrated in various muscle samples. On the same muscle samples which were analysed for mRNA expression, the corresponding MyHC proteins were studied with SDS PAGE and Western blot. The mRNA-protein comparison allowed the identification of 10 distinct proteins based on the electrophoretic migration rate. Three groups were formed based on the migration rate: fast migrating comprising beta/slow/1, alpha cardiac and fast 2B, slow migrating comprising fast 2X, fast 2A and two developmental isoforms (NEO and EMB), intermediate migrating comprising EO MyHC, slow B (product of MYH15), slow tonic (product of MYH14/7b). Of special interest was the demonstration of a protein band corresponding to 2B-MyHC in laryngeal muscles and the finding that all 10 isoforms are expressed in extraocular muscles. These latter muscles are the unique localization for extraocular, slow B (product of MYH15) and slow tonic (product of MYH14/7b).
authors.affiliation: Department of Comparative Biomedicine and Food Science, University of Padova, viale dell'Università 16, Legnaro, 35020 Padova, Italy. Electronic address: masca@unipd.it.
authors.forename: Francesco
authors.initials: F
authors.lastname: Mascarello
authors.affiliation: Department of Biomedical Sciences, University of Padova Via Marzolo 3, 35131 Padova, Italy.
authors.forename: Luana
authors.initials: L
authors.lastname: Toniolo
authors.affiliation: Department of Biomedical Sciences, University of Padova Via Marzolo 3, 35131 Padova, Italy.
authors.forename: Pasqua
authors.initials: P
authors.lastname: Cancellara
authors.affiliation: Department of Biomedical Sciences, University of Padova Via Marzolo 3, 35131 Padova, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Reggiani
authors.affiliation: Department of Comparative Biomedicine and Food Science, University of Padova, viale dell'Università 16, Legnaro, 35020 Padova, Italy.
authors.forename: Lisa
authors.initials: L
authors.lastname: Maccatrozzo
mesh_terms: D000818:Animals; D002415:Cats; D002417:Cattle; D004285:Dogs; D047108:Embryonic Development / Q000235:genetics*; D014644:Genetic Variation / Q000235:genetics*; D000882:Haplorhini; D006736:Horses; D006801:Humans; D051379:Mice; D018482:Muscle, Skeletal / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D012518:Sarcomeres / Q000235:genetics*; D013045:Species Specificity; D013552:Swine
chemical_list: D020033:Protein Isoforms; D018995:Myosin Heavy Chains
keywords: MyHCs expression; Preotical (extraocular) muscles; Presomitic muscles; Real Time PCR assay; SDS PAGE and Western blot; Somatic muscles