Measurement of insulin- and contraction-stimulated glucose uptake in isolated and incubated mature skeletal muscle from mice

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Standard

Measurement of insulin- and contraction-stimulated glucose uptake in isolated and incubated mature skeletal muscle from mice. / Kjøbsted, Rasmus; Kido, Kohei; Larsen, Jeppe Kjærgaard; Jørgensen, Nicolas Oldenburg; Birk, Jesper Bratz; Hellsten, Ylva; Wojtaszewski, Jørgen.

I: Journal of Visualized Experiments, Nr. 171, e61398, 2021.

Publikation: Bidrag til tidsskriftTidsskriftartikelfagfællebedømt

Harvard

Kjøbsted, R, Kido, K, Larsen, JK, Jørgensen, NO, Birk, JB, Hellsten, Y & Wojtaszewski, J 2021, 'Measurement of insulin- and contraction-stimulated glucose uptake in isolated and incubated mature skeletal muscle from mice', Journal of Visualized Experiments, nr. 171, e61398. https://doi.org/10.3791/61398

APA

Kjøbsted, R., Kido, K., Larsen, J. K., Jørgensen, N. O., Birk, J. B., Hellsten, Y., & Wojtaszewski, J. (2021). Measurement of insulin- and contraction-stimulated glucose uptake in isolated and incubated mature skeletal muscle from mice. Journal of Visualized Experiments, (171), [e61398]. https://doi.org/10.3791/61398

Vancouver

Kjøbsted R, Kido K, Larsen JK, Jørgensen NO, Birk JB, Hellsten Y o.a. Measurement of insulin- and contraction-stimulated glucose uptake in isolated and incubated mature skeletal muscle from mice. Journal of Visualized Experiments. 2021;(171). e61398. https://doi.org/10.3791/61398

Author

Kjøbsted, Rasmus ; Kido, Kohei ; Larsen, Jeppe Kjærgaard ; Jørgensen, Nicolas Oldenburg ; Birk, Jesper Bratz ; Hellsten, Ylva ; Wojtaszewski, Jørgen. / Measurement of insulin- and contraction-stimulated glucose uptake in isolated and incubated mature skeletal muscle from mice. I: Journal of Visualized Experiments. 2021 ; Nr. 171.

Bibtex

@article{6bc591d4e6bd4768918920a8bb7a2a76,
title = "Measurement of insulin- and contraction-stimulated glucose uptake in isolated and incubated mature skeletal muscle from mice",
abstract = "Skeletal muscle is an insulin-responsive tissue and typically takes up most of the glucose that enters the blood after a meal. Moreover, it has been reported that skeletal muscle may increase the extraction of glucose from the blood by up to 50-fold during exercise compared to resting conditions. The increase in muscle glucose uptake during exercise and insulin stimulation is dependent on the translocation of glucose transporter 4 (GLUT4) from intracellular compartments to the muscle cell surface membrane, as well as phosphorylation of glucose to glucose-6-phosphate by hexokinase II. Isolation and incubation of mouse muscles such as m. soleus and m. extensor digitorum longus (EDL) is an appropriate ex vivo model to study the effects of insulin and electrically-induced contraction (a model for exercise) on glucose uptake in mature skeletal muscle. Thus, the ex vivo model permits evaluation of muscle insulin sensitivity and makes it possible to match muscle force production during contraction ensuring uniform recruitment of muscle fibers during measurements of muscle glucose uptake. Moreover, the described model is suitable for pharmacological compound testing that may have an impact on muscle insulin sensitivity or may be of help when trying to delineate the regulatory complexity of skeletal muscle glucose uptake. Here we describe and provide a detailed protocol on how to measure insulin- and contraction-stimulated glucose uptake in isolated and incubated soleus and EDL muscle preparations from mice using radiolabeled [3H]2-deoxy-D-glucose and [14C]mannitol as an extracellular marker. This allows accurate assessment of glucose uptake in mature skeletal muscle in the absence of confounding factors that may interfere in the intact animal model. In addition, we provide information on metabolic viability of incubated mouse skeletal muscle suggesting that the method applied possesses some caveats under certain conditions when studying muscle energy metabolism.",
author = "Rasmus Kj{\o}bsted and Kohei Kido and Larsen, {Jeppe Kj{\ae}rgaard} and J{\o}rgensen, {Nicolas Oldenburg} and Birk, {Jesper Bratz} and Ylva Hellsten and J{\o}rgen Wojtaszewski",
note = "CURIS 2021 NEXS 187",
year = "2021",
doi = "10.3791/61398",
language = "English",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "Journal of Visualized Experiments",
number = "171",

}

RIS

TY - JOUR

T1 - Measurement of insulin- and contraction-stimulated glucose uptake in isolated and incubated mature skeletal muscle from mice

AU - Kjøbsted, Rasmus

AU - Kido, Kohei

AU - Larsen, Jeppe Kjærgaard

AU - Jørgensen, Nicolas Oldenburg

AU - Birk, Jesper Bratz

AU - Hellsten, Ylva

AU - Wojtaszewski, Jørgen

N1 - CURIS 2021 NEXS 187

PY - 2021

Y1 - 2021

N2 - Skeletal muscle is an insulin-responsive tissue and typically takes up most of the glucose that enters the blood after a meal. Moreover, it has been reported that skeletal muscle may increase the extraction of glucose from the blood by up to 50-fold during exercise compared to resting conditions. The increase in muscle glucose uptake during exercise and insulin stimulation is dependent on the translocation of glucose transporter 4 (GLUT4) from intracellular compartments to the muscle cell surface membrane, as well as phosphorylation of glucose to glucose-6-phosphate by hexokinase II. Isolation and incubation of mouse muscles such as m. soleus and m. extensor digitorum longus (EDL) is an appropriate ex vivo model to study the effects of insulin and electrically-induced contraction (a model for exercise) on glucose uptake in mature skeletal muscle. Thus, the ex vivo model permits evaluation of muscle insulin sensitivity and makes it possible to match muscle force production during contraction ensuring uniform recruitment of muscle fibers during measurements of muscle glucose uptake. Moreover, the described model is suitable for pharmacological compound testing that may have an impact on muscle insulin sensitivity or may be of help when trying to delineate the regulatory complexity of skeletal muscle glucose uptake. Here we describe and provide a detailed protocol on how to measure insulin- and contraction-stimulated glucose uptake in isolated and incubated soleus and EDL muscle preparations from mice using radiolabeled [3H]2-deoxy-D-glucose and [14C]mannitol as an extracellular marker. This allows accurate assessment of glucose uptake in mature skeletal muscle in the absence of confounding factors that may interfere in the intact animal model. In addition, we provide information on metabolic viability of incubated mouse skeletal muscle suggesting that the method applied possesses some caveats under certain conditions when studying muscle energy metabolism.

AB - Skeletal muscle is an insulin-responsive tissue and typically takes up most of the glucose that enters the blood after a meal. Moreover, it has been reported that skeletal muscle may increase the extraction of glucose from the blood by up to 50-fold during exercise compared to resting conditions. The increase in muscle glucose uptake during exercise and insulin stimulation is dependent on the translocation of glucose transporter 4 (GLUT4) from intracellular compartments to the muscle cell surface membrane, as well as phosphorylation of glucose to glucose-6-phosphate by hexokinase II. Isolation and incubation of mouse muscles such as m. soleus and m. extensor digitorum longus (EDL) is an appropriate ex vivo model to study the effects of insulin and electrically-induced contraction (a model for exercise) on glucose uptake in mature skeletal muscle. Thus, the ex vivo model permits evaluation of muscle insulin sensitivity and makes it possible to match muscle force production during contraction ensuring uniform recruitment of muscle fibers during measurements of muscle glucose uptake. Moreover, the described model is suitable for pharmacological compound testing that may have an impact on muscle insulin sensitivity or may be of help when trying to delineate the regulatory complexity of skeletal muscle glucose uptake. Here we describe and provide a detailed protocol on how to measure insulin- and contraction-stimulated glucose uptake in isolated and incubated soleus and EDL muscle preparations from mice using radiolabeled [3H]2-deoxy-D-glucose and [14C]mannitol as an extracellular marker. This allows accurate assessment of glucose uptake in mature skeletal muscle in the absence of confounding factors that may interfere in the intact animal model. In addition, we provide information on metabolic viability of incubated mouse skeletal muscle suggesting that the method applied possesses some caveats under certain conditions when studying muscle energy metabolism.

U2 - 10.3791/61398

DO - 10.3791/61398

M3 - Journal article

C2 - 34057444

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 171

M1 - e61398

ER -

ID: 270664259