Actin sliding velocity on pure myosin isoforms from hindlimb unloaded mice

M. Maffei, E. Longa, R. Qaisar, V. Agoni, J. F. Desaphy, D. Conte Camerino, R. Bottinelli, M. Canepari

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Aim: Notwithstanding the widely accepted idea that following disuse skeletal muscles become faster, an increase in shortening velocity was previously observed mostly in fibres containing type 1 myosin, whereas a decrease was generally found in fibres containing type 2B myosin. In this study, unloaded shortening velocity of pure type 1 and 2B fibres from hindlimb unloaded mice was determined and a decrease in type 2B fibres was found. Methods: To clarify whether the decrease in shortening velocity could depend on alterations of myosin motor function, an in vitro motility assay approach was applied to study pure type 1 and pure type 2B myosin from hindlimb unloaded mice. The latter approach, assessing actin sliding velocity on isolated myosin in the absence of other myofibrillar proteins, enabled to directly investigate myosin motor function. Results: Actin sliding velocity was significantly lower on type 2B myosin following unloading (2.70 ± 0.32 μm s-1) than in control conditions (4.11 ± 0.35 μm s-1), whereas actin sliding velocity of type 1 myosin was not different following unloading (0.89 ± 0.04 μm s-1) compared with control conditions (0.84 ± 0.17 μm s-1). Myosin light chain (MLC) isoform composition of type 2B myosin from hindlimb unloaded and control mice was not different. No oxidation of either type 1 or 2B myosin was observed. Higher phosphorylation of regulatory MLC in type 2B myosin after unloading was found. Conclusion: Results suggest that the observed lower shortening velocity of type 2B fibres following unloading could be related to slowing of acto-myosin kinetics in the presence of MLC phosphorylation.

Original languageEnglish
Pages (from-to)316-329
Number of pages14
JournalActa Physiologica
Volume212
Issue number4
DOIs
Publication statusPublished - Dec 1 2014

Fingerprint

Myosins
Hindlimb
Actins
Protein Isoforms
Myosin Light Chains
Myosin Type I
pioglitazone
Phosphorylation
Skeletal Muscle

Keywords

  • Disuse
  • Hindlimb unloading
  • in vitro motility assay
  • Myosin isoforms
  • Myosin light chain phosphorylation
  • Shortening velocity

ASJC Scopus subject areas

  • Physiology
  • Medicine(all)

Cite this

Maffei, M., Longa, E., Qaisar, R., Agoni, V., Desaphy, J. F., Camerino, D. C., ... Canepari, M. (2014). Actin sliding velocity on pure myosin isoforms from hindlimb unloaded mice. Acta Physiologica, 212(4), 316-329. https://doi.org/10.1111/apha.12320

Actin sliding velocity on pure myosin isoforms from hindlimb unloaded mice. / Maffei, M.; Longa, E.; Qaisar, R.; Agoni, V.; Desaphy, J. F.; Camerino, D. Conte; Bottinelli, R.; Canepari, M.

In: Acta Physiologica, Vol. 212, No. 4, 01.12.2014, p. 316-329.

Research output: Contribution to journalArticle

Maffei, M, Longa, E, Qaisar, R, Agoni, V, Desaphy, JF, Camerino, DC, Bottinelli, R & Canepari, M 2014, 'Actin sliding velocity on pure myosin isoforms from hindlimb unloaded mice', Acta Physiologica, vol. 212, no. 4, pp. 316-329. https://doi.org/10.1111/apha.12320
Maffei M, Longa E, Qaisar R, Agoni V, Desaphy JF, Camerino DC et al. Actin sliding velocity on pure myosin isoforms from hindlimb unloaded mice. Acta Physiologica. 2014 Dec 1;212(4):316-329. https://doi.org/10.1111/apha.12320
Maffei, M. ; Longa, E. ; Qaisar, R. ; Agoni, V. ; Desaphy, J. F. ; Camerino, D. Conte ; Bottinelli, R. ; Canepari, M. / Actin sliding velocity on pure myosin isoforms from hindlimb unloaded mice. In: Acta Physiologica. 2014 ; Vol. 212, No. 4. pp. 316-329.
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AU - Qaisar, R.

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AU - Desaphy, J. F.

AU - Camerino, D. Conte

AU - Bottinelli, R.

AU - Canepari, M.

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N2 - Aim: Notwithstanding the widely accepted idea that following disuse skeletal muscles become faster, an increase in shortening velocity was previously observed mostly in fibres containing type 1 myosin, whereas a decrease was generally found in fibres containing type 2B myosin. In this study, unloaded shortening velocity of pure type 1 and 2B fibres from hindlimb unloaded mice was determined and a decrease in type 2B fibres was found. Methods: To clarify whether the decrease in shortening velocity could depend on alterations of myosin motor function, an in vitro motility assay approach was applied to study pure type 1 and pure type 2B myosin from hindlimb unloaded mice. The latter approach, assessing actin sliding velocity on isolated myosin in the absence of other myofibrillar proteins, enabled to directly investigate myosin motor function. Results: Actin sliding velocity was significantly lower on type 2B myosin following unloading (2.70 ± 0.32 μm s-1) than in control conditions (4.11 ± 0.35 μm s-1), whereas actin sliding velocity of type 1 myosin was not different following unloading (0.89 ± 0.04 μm s-1) compared with control conditions (0.84 ± 0.17 μm s-1). Myosin light chain (MLC) isoform composition of type 2B myosin from hindlimb unloaded and control mice was not different. No oxidation of either type 1 or 2B myosin was observed. Higher phosphorylation of regulatory MLC in type 2B myosin after unloading was found. Conclusion: Results suggest that the observed lower shortening velocity of type 2B fibres following unloading could be related to slowing of acto-myosin kinetics in the presence of MLC phosphorylation.

AB - Aim: Notwithstanding the widely accepted idea that following disuse skeletal muscles become faster, an increase in shortening velocity was previously observed mostly in fibres containing type 1 myosin, whereas a decrease was generally found in fibres containing type 2B myosin. In this study, unloaded shortening velocity of pure type 1 and 2B fibres from hindlimb unloaded mice was determined and a decrease in type 2B fibres was found. Methods: To clarify whether the decrease in shortening velocity could depend on alterations of myosin motor function, an in vitro motility assay approach was applied to study pure type 1 and pure type 2B myosin from hindlimb unloaded mice. The latter approach, assessing actin sliding velocity on isolated myosin in the absence of other myofibrillar proteins, enabled to directly investigate myosin motor function. Results: Actin sliding velocity was significantly lower on type 2B myosin following unloading (2.70 ± 0.32 μm s-1) than in control conditions (4.11 ± 0.35 μm s-1), whereas actin sliding velocity of type 1 myosin was not different following unloading (0.89 ± 0.04 μm s-1) compared with control conditions (0.84 ± 0.17 μm s-1). Myosin light chain (MLC) isoform composition of type 2B myosin from hindlimb unloaded and control mice was not different. No oxidation of either type 1 or 2B myosin was observed. Higher phosphorylation of regulatory MLC in type 2B myosin after unloading was found. Conclusion: Results suggest that the observed lower shortening velocity of type 2B fibres following unloading could be related to slowing of acto-myosin kinetics in the presence of MLC phosphorylation.

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