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Extracellular Glucose Depletion as an Indirect Measure of Glucose Uptake in Cells and Tissues Ex Vivo
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Protocol to measure glucose utilization in mouse tissues using radiolabeled 2-deoxy-D-glucose
Cyun-Ming Wu1, Hao-Yun Li1, Wen-Yi Li2
1Department of Medicine, College of Medicine, National Taiwan University, Taipei 100, Taiwan.
STAR Protocols
|December 7, 2024
Summary
This study details a protocol for measuring glucose utilization in mice using radiolabeled 2-deoxy-D-glucose (2DG). The method quanties glucose metabolism in various tissues, offering insights into cellular energy processes.
Area of Science:
- Biochemistry
- Physiology
- Metabolic Research
Background:
- 2-deoxy-D-glucose (2DG) is a glucose analog.
- Hexokinase converts 2DG to 2DG-6P in glycolysis.
- 2DG measures glucose uptake, while 2DG-6P indicates glucose utilization.
Purpose of the Study:
- To present a protocol for measuring glucose utilization in mouse tissues.
- To quantify metabolic activity using radiolabeled 2DG.
- To establish a method for assessing tissue-specific glucose metabolism.
Main Methods:
- Preparation of mice and chemical reagents.
- Blood extraction and chemical administration.
- Tissue dissolution and radioactivity measurement.
- Calculation of glucose utilization via the trapezoid rule.
Main Results:
- A detailed protocol for measuring glucose utilization in vivo.
- Demonstration of quantifying 2DG-6P formation in various tissues.
- Method allows for assessment of metabolic flux.
Conclusions:
- The presented protocol enables accurate measurement of glucose utilization in mouse tissues.
- This method provides a valuable tool for studying metabolic pathways.
- The findings contribute to understanding glucose metabolism and its alterations in disease.
