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An optimized method for tissue glycogen quantification

Mobilization of glycogen, the short‐term storage form of glucose, is the body's first defense against hypoglycemia and is critical for energy provision during high intensity exercise. Therefore, to advance metabolic research, it is critical to be able to accurately measure glycogen concentratio...

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Autores principales: Schaubroeck, Kyle J., Leitner, Brooks P., Perry, Rachel J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8855679/
https://www.ncbi.nlm.nih.gov/pubmed/35179318
http://dx.doi.org/10.14814/phy2.15195
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author Schaubroeck, Kyle J.
Leitner, Brooks P.
Perry, Rachel J.
author_facet Schaubroeck, Kyle J.
Leitner, Brooks P.
Perry, Rachel J.
author_sort Schaubroeck, Kyle J.
collection PubMed
description Mobilization of glycogen, the short‐term storage form of glucose, is the body's first defense against hypoglycemia and is critical for energy provision during high intensity exercise. Therefore, to advance metabolic research, it is critical to be able to accurately measure glycogen concentrations, including during a prolonged fast and other glycogen‐modulating interventions. Unfortunately, prior enzymatic methods of glycogen detection have been plagued by poor detection in the lower range, high sample mass requirements, and complicated and/or expensive protocols which introduce substantial technical variability into the measured glycogen concentrations. To address these limitations, here we report a streamlined and versatile glycogen extraction protocol coupled with an optimized phenol‐sulfuric acid quantification protocol. With this method, we demonstrate how glycogen can be extracted from only 20 mg of tissue with one centrifugation step and quantified with a highly precise (Intra‐assay %CV ranges from 5–10%) and sensitive (proportionality constant for glycogen = 0.07279 A.U./µg) assay. The cost of all materials equates to ~10 cents per sample. Therefore, this method represents an attractive means of assessing ex vivo tissue glycogen content including at the extremes of glycogen concentrations.
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spelling pubmed-88556792022-02-25 An optimized method for tissue glycogen quantification Schaubroeck, Kyle J. Leitner, Brooks P. Perry, Rachel J. Physiol Rep Original Articles Mobilization of glycogen, the short‐term storage form of glucose, is the body's first defense against hypoglycemia and is critical for energy provision during high intensity exercise. Therefore, to advance metabolic research, it is critical to be able to accurately measure glycogen concentrations, including during a prolonged fast and other glycogen‐modulating interventions. Unfortunately, prior enzymatic methods of glycogen detection have been plagued by poor detection in the lower range, high sample mass requirements, and complicated and/or expensive protocols which introduce substantial technical variability into the measured glycogen concentrations. To address these limitations, here we report a streamlined and versatile glycogen extraction protocol coupled with an optimized phenol‐sulfuric acid quantification protocol. With this method, we demonstrate how glycogen can be extracted from only 20 mg of tissue with one centrifugation step and quantified with a highly precise (Intra‐assay %CV ranges from 5–10%) and sensitive (proportionality constant for glycogen = 0.07279 A.U./µg) assay. The cost of all materials equates to ~10 cents per sample. Therefore, this method represents an attractive means of assessing ex vivo tissue glycogen content including at the extremes of glycogen concentrations. John Wiley and Sons Inc. 2022-02-18 /pmc/articles/PMC8855679/ /pubmed/35179318 http://dx.doi.org/10.14814/phy2.15195 Text en © 2022 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Schaubroeck, Kyle J.
Leitner, Brooks P.
Perry, Rachel J.
An optimized method for tissue glycogen quantification
title An optimized method for tissue glycogen quantification
title_full An optimized method for tissue glycogen quantification
title_fullStr An optimized method for tissue glycogen quantification
title_full_unstemmed An optimized method for tissue glycogen quantification
title_short An optimized method for tissue glycogen quantification
title_sort optimized method for tissue glycogen quantification
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8855679/
https://www.ncbi.nlm.nih.gov/pubmed/35179318
http://dx.doi.org/10.14814/phy2.15195
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