title: Absolute quantification of myosin heavy chain isoforms by selected reaction monitoring can underscore skeletal muscle changes in a mouse model of amyotrophic lateral sclerosis.
abstract: Skeletal muscle fibers contain different isoforms of myosin heavy chain (MyHC) that define distinctive contractile properties. In light of the muscle capacity to adapt MyHC expression to pathophysiological conditions, a rapid and quantitative assessment of MyHC isoforms in small muscle tissue quantities would represent a valuable diagnostic tool for (neuro)muscular diseases. As past protocols did not meet these requirements, in the present study we applied a targeted proteomic approach based on selected reaction monitoring that allowed the absolute quantification of slow and fast MyHC isoforms in different mouse skeletal muscles with high reproducibility. This mass-spectrometry-based method was validated also in a pathological specimen, by comparison of the MyHC expression profiles in different muscles from healthy mice and a genetic mouse model of amyotrophic lateral sclerosis (ALS) expressing the SOD1(G93A) mutant. This analysis showed that terminally ill ALS mice have a fast-to-slow shift in the fiber type composition of the tibialis anterior and gastrocnemius muscles, as previously reported. These results will likely open the way to accurate and rapid diagnoses of human (neuro)muscular diseases by the proposed method. Graphical Abstract Methods for myosin heavy chain (MyHC) quantification: a comparison of classical methods and selected reaction monitoring (SRM)-based mass spectrometry approaches.
authors.affiliation: Department of Biomedical Sciences, University of Padua, Via Ugo Bassi 58/B, 35131, Padua, PD, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Peggion
authors.affiliation: CNR Neuroscience Institute, University of Padua, Via Ugo Bassi 58/B, 35131, Padua, PD, Italy.
authors.forename: Maria Lina
authors.initials: ML
authors.lastname: Massimino
authors.affiliation: Department of Chemistry, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università, 10, 35020, Legnaro, PD, Italy.
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Biancotto
authors.affiliation: Department of Chemistry, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università, 10, 35020, Legnaro, PD, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Angeletti
authors.affiliation: Department of Biomedical Sciences, University of Padua, Via Ugo Bassi 58/B, 35131, Padua, PD, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Reggiani
authors.affiliation: Department of Biomedical Sciences, University of Padua, Via Ugo Bassi 58/B, 35131, Padua, PD, Italy.|CNR Neuroscience Institute, University of Padua, Via Ugo Bassi 58/B, 35131, Padua, PD, Italy.
authors.forename: Maria Catia
authors.initials: MC
authors.lastname: Sorgato
authors.affiliation: Department of Biomedical Sciences, University of Padua, Via Ugo Bassi 58/B, 35131, Padua, PD, Italy. alessandro.bertoli@unipd.it.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Bertoli
authors.affiliation: Department of Chemistry, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università, 10, 35020, Legnaro, PD, Italy. rstella@izsvenezie.it.
authors.forename: Roberto
authors.initials: R
authors.lastname: Stella
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000378:metabolism / Q000473:pathology*; D000818:Animals; D004195:Disease Models, Animal; D006801:Humans; D051379:Mice; D018482:Muscle, Skeletal / Q000473:pathology*; D018995:Myosin Heavy Chains / Q000032:analysis*
chemical_list: D018995:Myosin Heavy Chains
keywords: Absolute quantification; Amyotrophic lateral sclerosis; Myosin heavy chain; Selected reaction monitoring; Skeletal muscle; Targeted proteomics