Mice with Whole-Body Disruption of AMPK-Glycogen Binding Have Increased Adiposity, Reduced Fat Oxidation and Altered Tissue Glycogen Dynamics
Abstract
:1. Introduction
2. Results
2.1. DKI Mice Display Increased Body Mass Associated with Increased Adiposity
2.2. DKI Mice Have Reduced Voluntary Physical Activity, Increased Rates of CHO Oxidation, and Reduced Rates of Fat Oxidation
2.3. DKI Mice Display Hyperinsulinemia and Glucose Intolerance but Normal Whole-Body Insulin Sensitivity
2.4. DKI Mice Have Decreased Voluntary Ambulatory Activity and Decreased Rates of Fat Oxidation during Overnight Fasting
2.5. DKI Mice Display Reduced Liver Glycogen Concentration in the Fed State and Altered Fasting-Induced Skeletal Muscle Glycogen Depletion
2.6. Liver and Muscle from DKI Mice Display No Changes in Glycogen Associated Proteins Relative to WT
2.7. DKI Mice Have Intact AMPK and ACC Phosphorylation in Liver and Skeletal Muscle
2.8. DKI Mice Have Reduced AMPK Protein Content in Liver and Skeletal Muscle
3. Discussion
4. Materials and Methods
4.1. Animal Models
4.2. Whole-Body Composition Measurements
4.3. Metabolic Caging
4.4. Intraperitoneal Glucose Tolerance and Insulin Tolerance Testing
4.5. Fasting and Refeeding Protocol
4.6. Serum Analysis
4.7. Assessment of Tissue Glycogen Content
4.8. Immunoblotting
4.9. RNA Extraction and Gene Expression Analysis
4.10. Statistical Analyses
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Tissue | WT Tissue Mass (mg) | DKI Tissue Mass (mg) | p-Value |
---|---|---|---|
Pancreas | 120 ± 12.0 | 167 ± 16.5 * | 0.029 |
Heart | 159 ± 10.3 | 176 ± 11.6 | 0.27 |
Gastrocnemius | 306 ± 12.9 | 328 ± 17.1 | 0.11 |
Soleus | 16 ± 1.8 | 14 ± 1.8 | 0.35 |
Target | Primary Antibody | Incubation | Secondary Antibody |
---|---|---|---|
AMPK α p-T172 | CST 2531 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:5000 | |
AMPK α | CST 2532 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:5000 | |
AMPK β1/β2 | CST 4150 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:5000 | |
ACC p-S79 | CST 11818 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:5000 | |
ACC | CST 3662 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:5000 | |
Akt p-S473 | CST 9271 | 1.5 h | GAR CST 7074 |
1:1000 | Room temperature | 1:5000 | |
Akt | CST 4691 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:5000 | |
AS160 p-T642 | CST 4288 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:5000 | |
AS160 | Abcam ab24469 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:5000 | |
GLUT4 | CST 2213 | Overnight | GAM CST 7076 |
1:1000 | 4 °C | 1:2000 | |
GS p-S641 | CST 47043 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:5000 | |
GS | CST 3886 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:5000 | |
Glycogenin1 | Abcam ab11171 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:2000 | |
GBE1 | Abcam ab180596 | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:2000 | |
GP | Antibody generated by B.E.K. laboratory | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:2000 | |
GDE | Antibody generated by B.E.K. laboratory | Overnight | GAR CST 7074 |
1:1000 | 4 °C | 1:2000 |
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Janzen, N.R.; Whitfield, J.; Murray-Segal, L.; Kemp, B.E.; Hawley, J.A.; Hoffman, N.J. Mice with Whole-Body Disruption of AMPK-Glycogen Binding Have Increased Adiposity, Reduced Fat Oxidation and Altered Tissue Glycogen Dynamics. Int. J. Mol. Sci. 2021, 22, 9616. https://doi.org/10.3390/ijms22179616
Janzen NR, Whitfield J, Murray-Segal L, Kemp BE, Hawley JA, Hoffman NJ. Mice with Whole-Body Disruption of AMPK-Glycogen Binding Have Increased Adiposity, Reduced Fat Oxidation and Altered Tissue Glycogen Dynamics. International Journal of Molecular Sciences. 2021; 22(17):9616. https://doi.org/10.3390/ijms22179616
Chicago/Turabian StyleJanzen, Natalie R., Jamie Whitfield, Lisa Murray-Segal, Bruce E. Kemp, John A. Hawley, and Nolan J. Hoffman. 2021. "Mice with Whole-Body Disruption of AMPK-Glycogen Binding Have Increased Adiposity, Reduced Fat Oxidation and Altered Tissue Glycogen Dynamics" International Journal of Molecular Sciences 22, no. 17: 9616. https://doi.org/10.3390/ijms22179616