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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

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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.

Methods for myosin heavy chain (MyHC) quantification: a comparison of classical methods and selected reaction monitoring (SRM)-based mass spectrometry approaches

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Acknowledgements

This work was supported by grants from the AriSLA Foundation (project LoCaLS 2013 to A.B.) and the University of Padua (PRAT CPDA121988/12 to M.C.S. and PRAT CPDA158035/15 to A.B.). The authors thank Giorgio Arrigoni and Marta Murgia (Department of Biomedical Science, University of Padua) for helpful discussions and advice.

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Authors and Affiliations

  1. Department of Biomedical Sciences, University of Padua, Via Ugo Bassi 58/B, 35131, Padua, PD, Italy

    Caterina Peggion, Carlo Reggiani, Maria Catia Sorgato & Alessandro Bertoli

  2. CNR Neuroscience Institute, University of Padua, Via Ugo Bassi 58/B, 35131, Padua, PD, Italy

    Maria Lina Massimino & Maria Catia Sorgato

  3. Department of Chemistry, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell’Università, 10, 35020, Legnaro, PD, Italy

    Giancarlo Biancotto, Roberto Angeletti & Roberto Stella

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  1. Caterina Peggion
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  2. Maria Lina Massimino
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  3. Giancarlo Biancotto
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  4. Roberto Angeletti
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  5. Carlo Reggiani
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  7. Alessandro Bertoli
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Correspondence to Alessandro Bertoli or Roberto Stella.

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Peggion, C., Massimino, M.L., Biancotto, G. et al. Absolute quantification of myosin heavy chain isoforms by selected reaction monitoring can underscore skeletal muscle changes in a mouse model of amyotrophic lateral sclerosis. Anal Bioanal Chem 409, 2143–2153 (2017). https://doi.org/10.1007/s00216-016-0160-2

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  • DOI: https://doi.org/10.1007/s00216-016-0160-2

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