Cold Induced Depot-Specific Browning in Ferret Aortic Perivascular Adipose Tissue

Figures

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  FIGURE 1. Uncoupling protein 1 immunostaining in tPVAT of ferrets acclimatized to different room temperatures: 22°C (control group) or 4°C (cold group) for 1 week. A strong browning induction in cold-exposed ferrets is observed, characterized by increased appearance of adipocytes positive for UCP1 protein. A representative image of an animal of each group is represented. For each image, magnifications of 25×, 200×, and 630× are shown. A, aorta; L, lymph node.

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  FIGURE 2. Uncoupling protein 1 immunostaining in IAT of ferrets acclimatized to different room temperatures: 22°C (control group) or 4°C (cold group) for 1 week. No appearance of UCP1 positive cells was observed as result of cold exposure. A representative image of an animal of each group is represented. For each image, magnifications of 25×, 200×, and 630× are shown.

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  FIGURE 3. (A) Schematic overview of regulated genes in tPVAT and in IAT in control (22°C) vs. cold (4°C) exposed ferrets (Student’s t-test, p-value < 0.01, with an absolute fold change ≥ 2). The arrows show the number of genes up- and down-regulated in each analysis. (B) PCA analysis of gene expression in tPVAT and IAT at 22 and 4°C of a subset of genes being differentially expressed in tPVAT vs. IAT at control (22°C) temperature (Student’s t-test, p-value < 0.01, with an absolute fold change ≥ 2). Volcano plot for all expressed probes in control (C) and cold conditions (D). Minus log10 p-value of all genes are plotted against the differences of group means (log ratio) of each gene, tPVAT vs. IAT. The line indicates the threshold p-value < 0.01. The results represent data from the control (n = 7) and cold (n = 6–7) groups.

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  FIGURE 4. Volcano Plot for all expressed probes by microarray analysis in tPVAT (A) and IAT (C). Minus log10 p-value of all genes are plotted against the differences of group means (log ratio) of each gene, cold vs. control. The line indicates the threshold p-value < 0.05. Detailed manual classification of the 50 top up- and down-regulated genes based on their fold change (Student’s t-test, p-value < 0.05) in tPVAT (B) and in IAT (D). The results represent data from the control (n = 7) and cold (n = 6–7) groups.

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  FIGURE 5. Schematic overview of gene expression regulation of fatty acid oxidation and tricarboxylic acid (TCA) cycle in tPVAT and IAT. Up-regulated genes are marked in black and down-regulated are marked in gray (Student’s t-test, p-value < 0.05). Source: WikiPathways (adapted). According to the license terms of WikiPathways, users are free to use the pathway images in presentations, documents, websites and publications; as well as to use the pathway data in analyses, qualitative or quantitative, while citing or giving appropriate attribution (https://www.wikipathways.org/index.php/WikiPathways:License_Terms). ACADL, Acyl-CoA Dehydrogenase, Long Chain; ACADM, Acyl-CoA Dehydrogenase, C-4 To C-12 Straight Chain; ACADS, Acyl-CoA Dehydrogenase, C-2 To C-3 Short Chain; ACADVL, Acyl-CoA Dehydrogenase, Very Long Chain; ACO-2, Aconitase; CPT1, Carnitine Palmitoyltransferase 1; CPT2, Carnitine Palmitoyltransferase 2; CS, Citrate Synthase; DLD, Dihydrolipoamide Dehydrogenase; EHHADH, Enoyl-CoA Hydratase And 3-Hydroxyacyl CoA; FFA, Free Fatty Acids; FABP3, Fatty Acid Binding Protein 3; FH, Fumarate Hydratase; HADH, Hydroxyacyl-CoA Dehydrogenase; HADHA, Hydroxyacyl-CoA Dehydrogenase/3-Ketoacyl-CoA;…