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

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 de...

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Bibliographic Details
Published in:Life science alliance 2023-10, Vol.6 (10), p.e202302227
Main Authors: Hennig, Katharina, Hardman, David, Barata, David Mb, Martins, Inês Ibb, Bernabeu, Miguel O, Gomes, Edgar R, Roman, William
Format: Article
Language:English
Subjects:
Muscle Fibers, Fast-Twitch - chemistry
Muscle Fibers, Fast-Twitch - metabolism
Muscle Fibers, Skeletal
Muscle Fibers, Slow-Twitch - chemistry
Muscle Fibers, Slow-Twitch - metabolism
Muscle, Skeletal - metabolism
Optogenetics
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Summary: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.
ISSN:2575-1077
2575-1077
DOI:10.26508/lsa.202302227