Mitochondrial pyruvate carrier is required for optimal brown fat thermogenesis
- Department of Biochemistry, University of Utah, United States
- Department of Nutrition and Integrative Physiology, University of Utah, United States
- Howard Hughes Medical Institute, University of Utah, United States
- Department of Integrative Biology and Physiology, University of California, Los Angeles, United States
Figures
Figure 1 with 1 supplement
Transcriptome and metabolomics analysis of brown fat shows increased carbohydrate metabolism and glycolytic metabolism during cold exposure.
(a) Network visualization of enriched biological pathways altered with cold exposure in BAT (N = 5). (b) GSEA pathway analysis of differentially expressing genes (FDR < 0.05) in BAT at 4°C versus …
Figure 1—figure supplement 1
RNRNA-seq analysis of brown adipose tissue from mice at room temperature or with cold exposure.
(a) Hierarchical clustering illustrates large-scale differences in gene expression between cold room and room temperature exposed BAT. (b) Unsupervised principal component analysis (PCA) shows that …
Figure 2 with 1 supplement
CL-316,243 stimulation of brown adipocytes leads to increased 13C-glucose flux.
(a) Atom mapping for [U-13C] glucose tracing into glycolysis and the TCA cycle. White balls are 12C atoms. Black balls are 13C atoms. (b) Tracing analysis from U-13C glucose in differentiated brown …
Figure 2—figure supplement 1
Metabolite tracing of brown adipocytes in response to β3-Adrenergic Receptor agonist.
(a) 13C labeling of M+6 glucose and M+three pyruvate and lactate in the media of brown adipocytes treated with vehicle or 100 nM CL-316,243 for 1 or 5 hours (N = 3). (b) Total intracellular pyruvate …
Figure 3 with 1 supplement
Loss of MPC1 in BAT impairs thermogenesis and leads to cold sensitivity.
(a) Relative gene expression in brown adipose tissue from mice adapted to 30°C or 4°C for 1 day or 1 week. N = 4–5. (b) Relative gene expression in brown adipose tissue from mice adapted to 30°C or …
Figure 3—figure supplement 1
Regulation of Mpc1 in response to the cold and the conditional deletion of Mpc1 in brown fat.
(a) Quantification of western blot analysis of brown adipose tissue of mice adapted to 30°C or 4°C for 1 week (N = 4–5), (p<0.05). (b) Quantification of western blot analysis of brown adipose tissue …
Figure 4 with 1 supplement
Conditional deletion of Mpc1 in BAT impairs systemic glucose metabolism.
(a) Glucose tolerance test at room temperature (23°C) and cold (4°C) in Mpc1F/F and Mpc1F/F::Ucp1Cre3–4 months old, N = 5. (b) Insulin tolerance test at room temperature (23°C) in Mpc1F/F and Mpc1F/F…
Figure 4—figure supplement 1
Conditional deletion of Mpc1 in BAT and the impact on body composition and blood glucose.
(a) Body composition of Mpc1F/F and Mpc1F/F::Ucp1Cre mice housed at room temperature by NMR. (b) Time-dependent changes in blood glucose levels in Mpc1F/F and Mpc1F/F::Ucp1Cre mice mice 3–4 months …
Figure 5 with 1 supplement
Loss of mitochondrial pyruvate carrier does not affect CL-316,243-stimulated increase in 13C-glycolytic flux.
(a) Western blot analysis of differentiated brown Mpc1F/F adipocytes expressing pMSCV2 or CreERT2 treated with ethanol or 4-hydroxy tamoxifen. N = 3. (b) Gene expression analysis in differentiated …
Figure 5—figure supplement 1
Isotopic labeling of glycolitic intermediates in brown adiopcytes labeled with [U-13C]-glucose.
(a) Labeling pattern of [U-13C]-glucose incorporation into the glycolytic intermediates in MPC1F/F adipocytes expressing CreERT2 treated with ethanol or 4-hydroxy tamoxifen, with/without 100 nM …
Figure 6 with 1 supplement
Mitochondrial pyruvate transport is required for 13C-glucose-derived TCA cycle intermediates.
(a) Atom mapping for [U-13C]-glucose tracing incorporation into the TCA cycle intermediates. White circles are 12C atoms. Black circles are 13C atoms. (b) [U-13C]-glucose labeling in Mpc1F/F …
Figure 6—figure supplement 1
Loss of Mpc1 in brown adipocytes impairs incorporation of glucose-derived TCA-cycle intermediates.
(a) Labeling pattern of [U-13C]-glucose incorporation into the TCA-cycle intermediates in Mpc1F/F adipocytes expressing CreERT2 treated with ethanol or 4-hydroxy tamoxifen, with/without 100 nM …
Figure 7 with 1 supplement
Conditional deletion of MPC1 in brown fat leads to compensatory increase in fatty acid oxidation.
(a) U-13C palmitate-tracing experiments in Mpc1F/F cells expressing CRE-ERT2. Cells were treated with ethanol or 4-hydroxy tamoxifen, with/without 100 nM CL-316,243 for 5 hours (N = 6). (b) Oxygen …
Figure 7—figure supplement 1
Proton leak in response to UK5099, etomoxir, or both.
(a) Seahorse analysis of proton leak in differentiated brown adipocytes treated with/without 100 nM CL-316,243 ± vehicle, 10 µM UK5099, 10 µM Etomoxir, or both (N = 10–12).
Figure 8 with 1 supplement
Conditional deletion of Mpc1 in brown fat leads to increased ketogenesis.
(a) Volcano plot showing changes in serum metabolites between Mpc1F/F and Mpc1F/F::Ucp1Cre mice housed at 4°C for 6 hr. N = 6. (b) Heat map of top 25 metabolites in BAT from Mpc1F/F and Mpc1F/F::Ucp1…
Figure 8—figure supplement 1
Gene expression analysis and serum FFA levels.
(a) Gene expression in liver from Mpc1F/F and Mpc1F/F::Ucp1Cre mice housed at 4°C for 6 hr. Data was analyzed by Student’s t-test. Values are shown as mean + s.e.m. (p<0.05). (b) Free fatty acid …
Additional files
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Supplementary file 1
List of 1907 genes up-regulated in response to the cold.
- https://cdn.elifesciences.org/articles/52558/elife-52558-supp1-v2.xlsx
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Supplementary file 2
List of 3273 genes down-regulated in response to the cold.
- https://cdn.elifesciences.org/articles/52558/elife-52558-supp2-v2.xlsx
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Supplementary file 3
Metabolite analysis in brown adipose tissue at 30°C, 23°C, and 4°C.
- https://cdn.elifesciences.org/articles/52558/elife-52558-supp3-v2.xlsx
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Supplementary file 4
Metabolite analysis of serum at 30°C, 23°C, and 4°C.
- https://cdn.elifesciences.org/articles/52558/elife-52558-supp4-v2.xlsx
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Supplementary file 5
List of metabolites from serum of MPC1 F/F and MPC1 F/F::UCP1-Cre mice housed at 4°C for 6 hr.
- https://cdn.elifesciences.org/articles/52558/elife-52558-supp5-v2.xlsx
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Supplementary file 6
List of metabolites from BAT of MPC1 F/F and MPC1 F/F UCP1 Cre mice housed at 4°C for 6 hr.
- https://cdn.elifesciences.org/articles/52558/elife-52558-supp6-v2.xlsx
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Supplementary file 7
Real-time PCR primer list.
- https://cdn.elifesciences.org/articles/52558/elife-52558-supp7-v2.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/52558/elife-52558-transrepform-v2.docx
