Comprehensive Analysis of 13C6 Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts
Abstract
:1. Introduction
2. Results
2.1. An Upregulated Pentose Phosphate Pathway (PPP) in Spalax Cells Provides Higher Levels of NADPH
2.2. Spalax Cells Branched Much More Consumed Glucose to Hexosamine Biosynthetic Pathway (HBP), but Not to Hyaluronic Acid Production
2.3. Spalax Cells Massively Redirect Glucose Carbons to the Synthesis of the Tripeptide Glu-Cys-Gly, Glutathione (GSH)
2.4. Upregulated Lactate Production in Spalax Cells Maintains NAD+ Pool for Glycolytic Machinery and Rejects Glucose-Derived Carbons from Downstream Processing in the TCA Cycle
2.5. Spalax Cells Metabolized Less Glc-Originated Pyruvate Entering via Pyruvate Dehydrogenase Complex (PDC) Than the Rat Cells, While Pyruvate Carboxylase (PC)-Mediated Anaplerosis Is More Pronounced in Spalax
2.6. Succinate Is Essential for Hypoxic Spalax Cells
2.7. A Large Part of Glc-Originated Carbons Was Diverted to the Metabolism along with the Axis αKG→Glu→Pro→HPro Instead of Processing in the TCA Cycle Reactions
2.8. Spalax Cells Possess Upregulated Flux of PC-Metabolized Glc Carbons into 2 Hydroxyglutarate (2HG)
2.9. The Newly-Produced Ac-CoA Forwards to Acetyl Carnitine (ALCAR) Synthesis More Than to De-Novo Fatty Acids Production in Spalax Cells
2.10. Hypoxic Spalax Cells Significantly Amplified the Harboring of Glc-Derived Carbons That Entered to Mitochondria as HPro
2.11. Spalax Cells Express Higher Levels of Hypoxia- Inducible Factor-1a (HIF-1α) under Normoxia and Hypoxia Compared to the Rat Cells
2.12. Estimation of the Mitochondrial Aerobic Metabolism (MAM) and the Glycolytic Function (GF) in Spalax and Rat Cells Using the Seahorse Platform
3. Discussion
4. Methods
4.1. General Experimental Design
4.2. Animals
4.3. Cell Culture
4.4. Experiment Schedule, Extraction, and LC-MS Analysis
4.5. 13C6 Glucose Tracing Experiment Design
4.6. LC-MS Metabolomics Analysis
4.7. Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) Measurements
4.8. Hyaluronic Acid Assay
4.9. Western Blot
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Miskevich, D.; Chaban, A.; Dronina, M.; Abramovich, I.; Gottlieb, E.; Shams, I. Comprehensive Analysis of 13C6 Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts. Metabolites 2021, 11, 734. https://doi.org/10.3390/metabo11110734
Miskevich D, Chaban A, Dronina M, Abramovich I, Gottlieb E, Shams I. Comprehensive Analysis of 13C6 Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts. Metabolites. 2021; 11(11):734. https://doi.org/10.3390/metabo11110734
Chicago/Turabian StyleMiskevich, Dmitry, Anastasia Chaban, Maria Dronina, Ifat Abramovich, Eyal Gottlieb, and Imad Shams. 2021. "Comprehensive Analysis of 13C6 Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts" Metabolites 11, no. 11: 734. https://doi.org/10.3390/metabo11110734