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

Katharina Hennig; David Hardman; David MB Barata; Inês IBB Martins; Miguel O Bernabeu; Edgar R Gomes; William Roman

doi:10.26508/lsa.202302227

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

Katharina Hennig, David Hardman, David MB Barata, Inês IBB Martins, Miguel O Bernabeu, Edgar R Gomes, William Roman

Research output: Contribution to journal › Article › peer-review

Abstract / Description of output

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 up-regulation of neuron receptor genes. We observed an up-regulation in the expression of fast myosin heavy-chain isoforms, which induced a shift towards a fast-twitch phenotype. This long-term in vitro exercise strategy can be used to study fiber specification and refine muscle disease modelling.

Original language	English
Article number	e202302227
Number of pages	12
Journal	Life Science Alliance
Volume	6
Issue number	10
Early online date	7 Aug 2023
DOIs	https://doi.org/10.26508/lsa.202302227
Publication status	Published - 1 Oct 2023

Keywords / Materials (for Non-textual outputs)

Muscle Fibers, Fast-Twitch/chemistry
Muscle Fibers, Slow-Twitch/chemistry
Optogenetics
Muscle Fibers, Skeletal
Muscle, Skeletal/metabolism

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10.26508/lsa.202302227Licence: Creative Commons: Attribution (CC-BY)

Generating fast-twitch_HENNIG_DOA27072023_VOR_CC-BYFinal published version, 3.5 MBLicence: Creative Commons: Attribution (CC-BY)

https://www.life-science-alliance.org/content/6/10/e202302227Licence: Creative Commons: Attribution (CC-BY)

Building a 3D innervated and irrigated muscle on a chip - MyoChip
Bernabeu, M. O.
EU government bodies
1/11/18 → 30/04/23
Project: Research

Cite this

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title = "Generating fast-twitch myotubes in vitro with an optogenetic-based, quantitative contractility assay",

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 up-regulation of neuron receptor genes. We observed an up-regulation in the expression of fast myosin heavy-chain isoforms, which induced a shift towards a fast-twitch phenotype. This long-term in vitro exercise strategy can be used to study fiber specification and refine muscle disease modelling.",

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author = "Katharina Hennig and David Hardman and Barata, {David MB} and Martins, {In{\^e}s IBB} and Bernabeu, {Miguel O} and Gomes, {Edgar R} and William Roman",

note = "{\textcopyright} 2023 Hennig et al. Funding: We thank all partners of the MyoChip consortium and all members of the Edgar Gomes and the Claudio Franco laboratory. We thank Afonso Malheiro and Judite Costa for support and discussions. We thank Ant{\'o}nio Temudo, Ana Nascimento, Aida Lima, and Jose Rino from the Bioimaging facility at iMM for ´ imaging assistance. This project has received funding from the European Union{\textquoteright}s Horizon 2020 research and innovation program under grant agreement FET-OPEN No. 801423 and the European Research Council H2020- GA 810207-ARPCOMPLEXITY, the ERC-2022-POC2 n 101113328 and the Association Fran{\c c}aise contre les Myopathies (AFM). W Roman was supported by the Baker Foundation of Australia.",

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AU - Hennig, Katharina

AU - Hardman, David

AU - Barata, David MB

AU - Martins, Inês IBB

AU - Bernabeu, Miguel O

AU - Gomes, Edgar R

AU - Roman, William

N1 - © 2023 Hennig et al. Funding: We thank all partners of the MyoChip consortium and all members of the Edgar Gomes and the Claudio Franco laboratory. We thank Afonso Malheiro and Judite Costa for support and discussions. We thank António Temudo, Ana Nascimento, Aida Lima, and Jose Rino from the Bioimaging facility at iMM for ´ imaging assistance. This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement FET-OPEN No. 801423 and the European Research Council H2020- GA 810207-ARPCOMPLEXITY, the ERC-2022-POC2 n 101113328 and the Association Française contre les Myopathies (AFM). W Roman was supported by the Baker Foundation of Australia.

PY - 2023/10/1

Y1 - 2023/10/1

N2 - 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 up-regulation of neuron receptor genes. We observed an up-regulation in the expression of fast myosin heavy-chain isoforms, which induced a shift towards a fast-twitch phenotype. This long-term in vitro exercise strategy can be used to study fiber specification and refine muscle disease modelling.

AB - 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 up-regulation of neuron receptor genes. We observed an up-regulation in the expression of fast myosin heavy-chain isoforms, which induced a shift towards a fast-twitch phenotype. This long-term in vitro exercise strategy can be used to study fiber specification and refine muscle disease modelling.

KW - Muscle Fibers, Fast-Twitch/chemistry

KW - Muscle Fibers, Slow-Twitch/chemistry

KW - Optogenetics

KW - Muscle Fibers, Skeletal

KW - Muscle, Skeletal/metabolism

U2 - 10.26508/lsa.202302227

DO - 10.26508/lsa.202302227

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SN - 2575-1077

VL - 6

JO - Life Science Alliance

JF - Life Science Alliance

IS - 10

M1 - e202302227

ER -

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

Abstract / Description of output

Keywords / Materials (for Non-textual outputs)

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Projects

Building a 3D innervated and irrigated muscle on a chip - MyoChip

Cite this