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Absolute quantification of myosin heavy chain isoforms by selected reaction monitoring can underscore skeletal muscle changes in a mouse model of amyotrophic lateral sclerosis

Author:
Peggion, Caterina, Massimino, Maria Lina, Biancotto, Giancarlo, Angeletti, Roberto, Reggiani, Carlo, Sorgato, Maria Catia, Bertoli, Alessandro, Stella, Roberto
Source:
Analytical and bioanalytical chemistry 2017 v.409 no.8 pp. 2143-2153
ISSN:
1618-2642
Subject:
amyotrophic lateral sclerosis, animal models, diagnostic techniques, humans, mass spectrometry, mice, monitoring, muscles, muscular diseases, mutants, myosin heavy chains, myotubes, skeletal muscle
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
Agid:
5758880