Development of a high-throughput method for real-time assessment of cellular metabolism in intact long skeletal muscle fibre bundles

Li, Rui, Steyn, Frederik J., Stout, Michael B., Lee, Kevin, Cully, Tanya R., Calderon, Juan C. and Ngo, Shuyan T. (2016) Development of a high-throughput method for real-time assessment of cellular metabolism in intact long skeletal muscle fibre bundles. The Journal of Physiology, 594 24: 7197-7213. doi:10.1113/JP272988


Author Li, Rui
Steyn, Frederik J.
Stout, Michael B.
Lee, Kevin
Cully, Tanya R.
Calderon, Juan C.
Ngo, Shuyan T.
Title Development of a high-throughput method for real-time assessment of cellular metabolism in intact long skeletal muscle fibre bundles
Journal name The Journal of Physiology   Check publisher's open access policy
ISSN 0022-3751
1469-7793
Publication date 2016-01-01
Year available 2016
Sub-type Article (original research)
DOI 10.1113/JP272988
Open Access Status Not yet assessed
Volume 594
Issue 24
Start page 7197
End page 7213
Total pages 17
Place of publication Chichester, West Sussex United Kingdom
Publisher Wiley-Blackwell Publishing
Language eng
Abstract Metabolic dysfunction in skeletal muscle contributes to the aetiology and development of muscle diseases and metabolic diseases. As such, assessment of skeletal muscle cellular bioenergetics provides a powerful means to understand the role of skeletal muscle metabolism in disease and to identify possible therapeutic targets. Here, we developed a method that allows for the real-time assessment of cellular respiration in intact skeletal muscle fibre bundles obtained from the extensor digitorum longus (EDL) muscle of adult mice. Using this method, we assessed the contribution of ATP turnover and proton leak to basal mitochondrial oxygen consumption rate (OCR). Our data demonstrate that the mitochondria in EDL fibres are loosely coupled. Moreover, in the presence of carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), we show that palmitate exposure induced comparable peak OCR and higher total OCR in EDL fibre bundles when compared to pyruvate exposure, suggesting that fatty acids might be a more sustainable fuel source for skeletal muscle when mitochondria are driven to maximal respiration. Application of this method to EDL fibre bundles obtained from chronic high-fat diet fed mice revealed lower basal OCR and enhanced mitochondrial oxidation capacity in the presence of FCCP when compared to the chow-diet fed control mice. By using a 96-well microplate format, our method provides a flexible and efficient platform to investigate mitochondrial parameters of intact skeletal muscle fibres obtained from adult mice.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

 
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Created: Thu, 17 Nov 2016, 06:34:19 EST by Ms Shyuan Ngo on behalf of School of Biomedical Sciences