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Generating fast-twitch myotubes in vitro using an optogenetic-based, quantitative contractility assay

Katharina Hennig, David Hardman, David Barata, Inês Martins, Miguel O. Bernabeu, Edgar R. Gomes, William Roman
doi: https://doi.org/10.1101/2022.10.04.510824
Katharina Hennig
1Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Avenida Professor Egas Moniz, 1649-028, Lisboa, Portugal
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David Hardman
4Centre for Medical Informatics, Usher Institute, The University of Edinburgh, Edinburgh EH16 4UX, United Kingdom
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David Barata
1Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Avenida Professor Egas Moniz, 1649-028, Lisboa, Portugal
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Inês Martins
1Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Avenida Professor Egas Moniz, 1649-028, Lisboa, Portugal
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Miguel O. Bernabeu
4Centre for Medical Informatics, Usher Institute, The University of Edinburgh, Edinburgh EH16 4UX, United Kingdom
5The Bayes Centre, The University of Edinburgh, Edinburgh EH8 9BT, United Kingdom
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Edgar R. Gomes
1Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Avenida Professor Egas Moniz, 1649-028, Lisboa, Portugal
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  • For correspondence: william.roman@medicina.ulisboa.pt
William Roman
1Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Avenida Professor Egas Moniz, 1649-028, Lisboa, Portugal
2Australian Regenerative Medicine Institute, Monash University, Clayton, Australia
3Victoria Node, EMBL Australia, Clayton, Australia
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  • For correspondence: william.roman@medicina.ulisboa.pt
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Abstract

The composition of fiber types within skeletal muscle impacts the tissue’s physiological characteristics and susceptibility to disease and ageing. In vitro systems should therefore account for fiber type composition when modelling muscle conditions. To induce fiber specification in vitro, we designed a quantitative contractility assay based on optogenetics and particle image velocimetry. We submitted cultured myotubes to long-term intermittent light stimulation patterns and characterized their structural and functional adaptations. After several days of in vitro exercise, myotubes contract faster and are more resistant to fatigue. The enhanced contractile functionality was accompanied by advanced maturation such as increased width and upregulation of neuron receptor genes. We observed an upregulation in the expression of distinct myosin heavy chain isoforms (namely, neonatal-Myh8 and fast-Myh), which induced a shift towards a fast fiber phenotype. This long-term in vitro exercise strategy can be used to study fiber specification and refine muscle disease modelling.

Competing Interest Statement

The authors have declared no competing interest.

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Posted October 05, 2022.
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Generating fast-twitch myotubes in vitro using an optogenetic-based, quantitative contractility assay
Katharina Hennig, David Hardman, David Barata, Inês Martins, Miguel O. Bernabeu, Edgar R. Gomes, William Roman
bioRxiv 2022.10.04.510824; doi: https://doi.org/10.1101/2022.10.04.510824
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Generating fast-twitch myotubes in vitro using an optogenetic-based, quantitative contractility assay
Katharina Hennig, David Hardman, David Barata, Inês Martins, Miguel O. Bernabeu, Edgar R. Gomes, William Roman
bioRxiv 2022.10.04.510824; doi: https://doi.org/10.1101/2022.10.04.510824

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