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ANGPTL4 mediates shuttling of lipid fuel to brown adipose tissue during sustained cold exposure

Dijk, Wieneke, Heine, Markus, Vergnes, Laurent, Boon, Mariëtte R, Schaart, Gert, Hesselink, Matthijs KC, Reue, Karen, van Marken Lichtenbelt, Wouter D, Olivecrona, Gunilla, Rensen, Patrick CN, Heeren, Joerg, Kersten, Sander·2015·eLife
DOI 10.7554/elife.08428PMC PMC4709329
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Abstract

Brown adipose tissue (BAT) activation via cold exposure is increasingly scrutinized as a potential approach to ameliorate cardio-metabolic risk. Transition to cold temperatures requires changes in the partitioning of energy substrates, re-routing fatty acids to BAT to fuel non-shivering thermogenesis. However, the mechanisms behind the redistribution of energy substrates to BAT remain largely unknown. Angiopoietin-like 4 (ANGPTL4), a protein that inhibits lipoprotein lipase (LPL) activity, is highly expressed in BAT. Here, we demonstrate that ANGPTL4 is part of a shuttling mechanism that directs fatty acids derived from circulating triglyceride-rich lipoproteins to BAT during cold. Specifically, we show that cold markedly down-regulates ANGPTL4 in BAT, likely via activation of AMPK, enhancing LPL activity and uptake of plasma triglyceride-derived fatty acids. In contrast, cold up-regulates ANGPTL4 in WAT, abolishing a cold-induced increase in LPL activity. Together, our data indicate that ANGPTL4 is an important regulator of plasma lipid partitioning during sustained cold.

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Figures

Figure 1. ANGPTL4 expression in BAT is down-regulated upon sustained cold exposure. (A) Immunoblot for mouse ANGPTL4 and mouse LPL in lysates of kidney, spleen, heart, muscle, gonodal WAT, inguinal WAT and BAT of a C57BL/6J wild-type mouse. (B) Validation of anti-mANGPTL4 antibody in BAT lysates of Angptl4-/-, wild-type and Angptl4-Tg mice (left panel). Detection of glycosylated and non-glycosylated mANGPTL4 following treatment of BAT homogenate of a wild-type mouse with PGNase (right panel). (C) Immunofluorescent staining of UCP1 (upper panel; UCP1 = green, DAPI = blue), DAPI only (middle panel) and hANGPTL4 (lower panel; hANGPTL4 = green, DAPI = blue) in frozen sections (5 μm) of human BAT. (D) Angptl4 mRNA in T37i cells after 0, 3, 6 or 9 days of differentiation. (E) Angptl4 mRNA in BAT lysates of wild-type mice exposed to 4°C or 28°C for 1 or 10 days. (F) Immunoblot for ANGPTL4 in BAT homogenates of wild-type mice exposed to 4°C or 28°C for 10 days, following treatment with or without PGNase. (G) Lpl mRNA in BAT lysates of wild-type mice exposed to 4°C or 28°C for 1 or 10 days. * Statistically significant compared to control wells or compared to mice exposed to 28°C according to Student’s t-test (p<0.05). Error bars represent ± SEM. n = 8–10 mice per group. DOI: http://dx.doi.org/10.7554/eLife.08428.003
Figure 1. ANGPTL4 expression in BAT is down-regulated upon sustained cold exposure. (A) Immunoblot for mouse ANGPTL4 and mouse LPL in lysates of kidney, spleen, heart, muscle, gonodal WAT, inguinal WAT and BAT of a C57BL/6J wild-type mouse. (B) Validation of anti-mANGPTL4 antibody in BAT lysates of Angptl4-/-, wild-type and Angptl4-Tg mice (left panel). Detection of glycosylated and non-glycosylated mANGPTL4 following treatment of BAT homogenate of a wild-type mouse with PGNase (right panel). (C) Immunofluorescent staining of UCP1 (upper panel; UCP1 = green, DAPI = blue), DAPI only (middle panel) and hANGPTL4 (lower panel; hANGPTL4 = green, DAPI = blue) in frozen sections (5 μm) of human BAT. (D) Angptl4 mRNA in T37i cells after 0, 3, 6 or 9 days of differentiation. (E) Angptl4 mRNA in BAT lysates of wild-type mice exposed to 4°C or 28°C for 1 or 10 days. (F) Immunoblot for ANGPTL4 in BAT homogenates of wild-type mice exposed to 4°C or 28°C for 10 days, following treatment with or without PGNase. (G) Lpl mRNA in BAT lysates of wild-type mice exposed to 4°C or 28°C for 1 or 10 days. * Statistically significant compared to control wells or compared to mice exposed to 28°C according to Student’s t-test (p<0.05). Error bars represent ± SEM. n = 8–10 mice per group. DOI: http://dx.doi.org/10.7554/eLife.08428.003CC BY · PubMed Central
Figure 2. Down-regulation of ANGPTL4 in BAT upon sustained cold exposure affects plasma TG levels. (A) Schematic representation of cold exposure experiment with Angptl4-/-, wild-type and Angptl4-Tg mice. (B) Haematoxylin & Eosin staining on BAT sections (5 μm) of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (C) Food intake of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C for 10 days. (D) Weight gain of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C during 10 days. (E) BAT and WAT tissue weights and (F) body temperature of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (G) Elovl3 and Ucp1 mRNA expression levels of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (H) Plasma glucose, (I) plasma free fatty acids, and (J) plasma glycerol levels of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (K) Glycerol levels in medium of differentiated primary white adipocytes from Angptl4-/-, wild-type and Angptl4-Tg mice, serum-starved and treated with 10 μM isoproterenol for 3 hr. (J) plasma TG levels of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (K) Fast protein liquid chromatography (FPLC) on pooled plasma samples of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C for 10 days, followed by analysis of TG levels in all fractions. *Statistically significant compared to mice of equal genotype at 28°C or between groups as indicated by bars according to two-way ANOVA followed by a post-hoc Tukey HSD test (p<0.05). Error bars represent ± SEM. n = 8–10 mice per group. DOI: http://dx.doi.org/10.7554/eLife.08428.004
Figure 2. Down-regulation of ANGPTL4 in BAT upon sustained cold exposure affects plasma TG levels. (A) Schematic representation of cold exposure experiment with Angptl4-/-, wild-type and Angptl4-Tg mice. (B) Haematoxylin & Eosin staining on BAT sections (5 μm) of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (C) Food intake of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C for 10 days. (D) Weight gain of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C during 10 days. (E) BAT and WAT tissue weights and (F) body temperature of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (G) Elovl3 and Ucp1 mRNA expression levels of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (H) Plasma glucose, (I) plasma free fatty acids, and (J) plasma glycerol levels of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (K) Glycerol levels in medium of differentiated primary white adipocytes from Angptl4-/-, wild-type and Angptl4-Tg mice, serum-starved and treated with 10 μM isoproterenol for 3 hr. (J) plasma TG levels of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (K) Fast protein liquid chromatography (FPLC) on pooled plasma samples of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C for 10 days, followed by analysis of TG levels in all fractions. *Statistically significant compared to mice of equal genotype at 28°C or between groups as indicated by bars according to two-way ANOVA followed by a post-hoc Tukey HSD test (p<0.05). Error bars represent ± SEM. n = 8–10 mice per group. DOI: http://dx.doi.org/10.7554/eLife.08428.004CC BY · PubMed Central
Figure 3. Down-regulation of ANGPTL4 in BAT upon sustained cold exposure promotes an increase in BAT LPL activity. (A) Angptl4 mRNA in BAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (B) Immunoblot for ANGPTL4 and LPL in BAT homogenates from Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (C) Total LPL activity and (D) Lpl mRNA in BAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. * Statistically significant compared to mice of equal genotype at 28°C or between groups as indicated by bars, according to two-way ANOVA followed by a post-hoc Tukey HSD test (p<0.05). Error bars represent ± SEM. n = 8–10 mice per group. DOI: http://dx.doi.org/10.7554/eLife.08428.005
Figure 3. Down-regulation of ANGPTL4 in BAT upon sustained cold exposure promotes an increase in BAT LPL activity. (A) Angptl4 mRNA in BAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (B) Immunoblot for ANGPTL4 and LPL in BAT homogenates from Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (C) Total LPL activity and (D) Lpl mRNA in BAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. * Statistically significant compared to mice of equal genotype at 28°C or between groups as indicated by bars, according to two-way ANOVA followed by a post-hoc Tukey HSD test (p<0.05). Error bars represent ± SEM. n = 8–10 mice per group. DOI: http://dx.doi.org/10.7554/eLife.08428.005CC BY · PubMed Central
Figure 4. Down-regulation of ANGPTL4 in BAT upon sustained cold exposure promotes an increase in TRL-derived fatty acid uptake by BAT. (A,B) Plasma 3H (A) and 14C (B) activity in wild-type mice exposed to 4°C or 28°C for 10 days intravenously injected with VLDL-like particles labelled with glycerol tri[3H]oleate (TRL FA) and [14C]cholesteryl-oleate (TRL Chol). (C,D) Plasma 3H (C) and 14C (D) activity in Angptl4-/-, wild-type and Angptl4-Tg mice intravenously injected with VLDL-like emulsion particles labelled with glycerol tri[3H]oleate (TRL FA) and [14C]cholesteryl-oleate (TRL Chol), following exposure to 4°C for 10 days. (E,F) 3H activity (E) and 14C activity (F) in interscapular BAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days and intravenously injected with VLDL-like particles labelled with glycerol tri[3H]oleate (TRL FA) and [14C]cholesteryl-oleate (TRL Chol). (G,H) 14C and 3H activity in interscapular BAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days and intravenously injected with chylomicron-like particles labelled with glycerol tri[14C]oleate (TRL FA) (G) and [3H]cholesteryl-oleyloleate (TRL Chol). (I) Fluorescent image of uptake of intravenously injected QD-TRLs into BAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 9 days. Image was taken 12 min post-injection. n = 2 mice per group.* Statistically significant compared to mice of equal genotype at 28°C or between groups as indicated by bars, according to two-way ANOVA followed by a post-hoc Tukey HSD test (p<0.05). Error bars represent ± SEM. n = 7 mice per group, unless otherwise indicated. DOI: http://dx.doi.org/10.7554/eLife.08428.006
Figure 4. Down-regulation of ANGPTL4 in BAT upon sustained cold exposure promotes an increase in TRL-derived fatty acid uptake by BAT. (A,B) Plasma 3H (A) and 14C (B) activity in wild-type mice exposed to 4°C or 28°C for 10 days intravenously injected with VLDL-like particles labelled with glycerol tri[3H]oleate (TRL FA) and [14C]cholesteryl-oleate (TRL Chol). (C,D) Plasma 3H (C) and 14C (D) activity in Angptl4-/-, wild-type and Angptl4-Tg mice intravenously injected with VLDL-like emulsion particles labelled with glycerol tri[3H]oleate (TRL FA) and [14C]cholesteryl-oleate (TRL Chol), following exposure to 4°C for 10 days. (E,F) 3H activity (E) and 14C activity (F) in interscapular BAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days and intravenously injected with VLDL-like particles labelled with glycerol tri[3H]oleate (TRL FA) and [14C]cholesteryl-oleate (TRL Chol). (G,H) 14C and 3H activity in interscapular BAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days and intravenously injected with chylomicron-like particles labelled with glycerol tri[14C]oleate (TRL FA) (G) and [3H]cholesteryl-oleyloleate (TRL Chol). (I) Fluorescent image of uptake of intravenously injected QD-TRLs into BAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 9 days. Image was taken 12 min post-injection. n = 2 mice per group.* Statistically significant compared to mice of equal genotype at 28°C or between groups as indicated by bars, according to two-way ANOVA followed by a post-hoc Tukey HSD test (p<0.05). Error bars represent ± SEM. n = 7 mice per group, unless otherwise indicated. DOI: http://dx.doi.org/10.7554/eLife.08428.006CC BY · PubMed Central
Figure 4—figure supplement 1. (A,B) Schematic representation of clearance studies in Angptl4-/-, wild-type and Angptl4-Tg mice, consisting of 10 days of cold exposure or thermo-neutrality, followed by measurement of triglyceride clearance via injection of either radiolabelled VLDL-like particles or chylomicron-like particles. Mice were sacrificed 20 min post-injection to analyze the distribution of the radioactive labels. DOI: http://dx.doi.org/10.7554/eLife.08428.007
Figure 4—figure supplement 1. (A,B) Schematic representation of clearance studies in Angptl4-/-, wild-type and Angptl4-Tg mice, consisting of 10 days of cold exposure or thermo-neutrality, followed by measurement of triglyceride clearance via injection of either radiolabelled VLDL-like particles or chylomicron-like particles. Mice were sacrificed 20 min post-injection to analyze the distribution of the radioactive labels. DOI: http://dx.doi.org/10.7554/eLife.08428.007CC BY · PubMed Central
Figure 5. Up-regulation of ANGPTL4 in WAT upon sustained cold exposure suppresses WAT LPL activity and TRL-derived fatty acid uptake. (A) Total LPL activity levels and (B) Angptl4 mRNA in WAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (C) Immunoblot for ANGPTL4 and LPL in WAT homogenates of wild-type mice exposed to 4°C or 28°C for 10 days. (D) Lpl mRNA in WAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (E–H) Activity of 3H and 14C radiolabels in WAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days and intravenously injected with radiolabelled VLDL-like (E,G) and chylomicron-like particles (F,H). TRL FA uptake (E,F) reflects uptake of glycerol tri[3H/14C]oleate, whereas TRL Chol uptake (G,H) reflects uptake of the core labels [14C]cholesteryl-oleate or [3H]cholesteryl-oleyloleate. (I) Fluorescent image of uptake of intravenously injected QD-TRLs into WAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 9 days. Image was taken after perfusion of mice with PBS containing 50 IU/mL heparin and upon cryosectioning of tissues. n = 2 mice per group. * Statistically significant compared to mice of equal genotype at 28°C or between groups as indicated by bars, according to two-way ANOVA followed by a post-hoc Tukey HSD test (p<0.05). Error bars represent ± SEM. n = 7–10 mice per group, unless otherwise indicated. DOI: http://dx.doi.org/10.7554/eLife.08428.008
Figure 5. Up-regulation of ANGPTL4 in WAT upon sustained cold exposure suppresses WAT LPL activity and TRL-derived fatty acid uptake. (A) Total LPL activity levels and (B) Angptl4 mRNA in WAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (C) Immunoblot for ANGPTL4 and LPL in WAT homogenates of wild-type mice exposed to 4°C or 28°C for 10 days. (D) Lpl mRNA in WAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. (E–H) Activity of 3H and 14C radiolabels in WAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days and intravenously injected with radiolabelled VLDL-like (E,G) and chylomicron-like particles (F,H). TRL FA uptake (E,F) reflects uptake of glycerol tri[3H/14C]oleate, whereas TRL Chol uptake (G,H) reflects uptake of the core labels [14C]cholesteryl-oleate or [3H]cholesteryl-oleyloleate. (I) Fluorescent image of uptake of intravenously injected QD-TRLs into WAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 9 days. Image was taken after perfusion of mice with PBS containing 50 IU/mL heparin and upon cryosectioning of tissues. n = 2 mice per group. * Statistically significant compared to mice of equal genotype at 28°C or between groups as indicated by bars, according to two-way ANOVA followed by a post-hoc Tukey HSD test (p<0.05). Error bars represent ± SEM. n = 7–10 mice per group, unless otherwise indicated. DOI: http://dx.doi.org/10.7554/eLife.08428.008CC BY · PubMed Central
Figure 5—figure supplement 1. (A,B) Plasma ANGPTL4 levels in 10 obese (A) and 10 lean (B) individuals before and after exposure to a mild cold (16°C) for 48 hr (Wijers et al., 2010). Differences between mild cold and baseline were statistically significant in the obese group (paired Student’s t-test (p<0.05)). n = 10 individuals per group. DOI: http://dx.doi.org/10.7554/eLife.08428.009
Figure 5—figure supplement 1. (A,B) Plasma ANGPTL4 levels in 10 obese (A) and 10 lean (B) individuals before and after exposure to a mild cold (16°C) for 48 hr (Wijers et al., 2010). Differences between mild cold and baseline were statistically significant in the obese group (paired Student’s t-test (p<0.05)). n = 10 individuals per group. DOI: http://dx.doi.org/10.7554/eLife.08428.009CC BY · PubMed Central
Figure 5—figure supplement 2. (A) Total LPL activity levels, (B) Angptl4 mRNA, and (C) Lpl mRNA in inguinal WAT (iWAT) of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. *Statistically significant compared to mice of equal genotype at 28°C or between groups as indicated by bars, according to two-way ANOVA followed by a post-hoc Tukey HSD test (p<0.05). Error bars represent ± SEM. n = 7–10 mice per group. DOI: http://dx.doi.org/10.7554/eLife.08428.010
Figure 5—figure supplement 2. (A) Total LPL activity levels, (B) Angptl4 mRNA, and (C) Lpl mRNA in inguinal WAT (iWAT) of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days. *Statistically significant compared to mice of equal genotype at 28°C or between groups as indicated by bars, according to two-way ANOVA followed by a post-hoc Tukey HSD test (p<0.05). Error bars represent ± SEM. n = 7–10 mice per group. DOI: http://dx.doi.org/10.7554/eLife.08428.010CC BY · PubMed Central
Figure 6. Uptake of TRL-like particles in liver, spleen and muscle is not affected by Angptl4 genotype. (A) 3H and 14C activity in liver, spleen, muscle, inguinal WAT (iWAT) and subscapular BAT (sBAT) of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days and intravenously injected with VLDL-like emulsion particles labelled with glycerol tri[3H]oleate (TRL FA) and [14C]cholesteryloleate (TRL Chol). (B) 3H and 14C activity in liver, spleen, muscle, iWAT and sBAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days and intravenously injected with chylomicron-like particles labelled with glycerol tri[14C]oleate (TRL FA) and [3H]cholesteryl-oleyloleate (TRL Chol). *Statistically significant compared to values of wild-type mice according to Student’s t-test (p<0.05). Error bars represent ± SEM. n = 7 mice per group. DOI: http://dx.doi.org/10.7554/eLife.08428.011
Figure 6. Uptake of TRL-like particles in liver, spleen and muscle is not affected by Angptl4 genotype. (A) 3H and 14C activity in liver, spleen, muscle, inguinal WAT (iWAT) and subscapular BAT (sBAT) of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days and intravenously injected with VLDL-like emulsion particles labelled with glycerol tri[3H]oleate (TRL FA) and [14C]cholesteryloleate (TRL Chol). (B) 3H and 14C activity in liver, spleen, muscle, iWAT and sBAT of Angptl4-/-, wild-type and Angptl4-Tg mice exposed to 4°C or 28°C for 10 days and intravenously injected with chylomicron-like particles labelled with glycerol tri[14C]oleate (TRL FA) and [3H]cholesteryl-oleyloleate (TRL Chol). *Statistically significant compared to values of wild-type mice according to Student’s t-test (p<0.05). Error bars represent ± SEM. n = 7 mice per group. DOI: http://dx.doi.org/10.7554/eLife.08428.011CC BY · PubMed Central
Figure 7. AMPK is activated in BAT, but not WAT, upon sustained cold exposure. (A) Angptl4 mRNA in differentiated mouse white adipocytes (primary adipocytes and 3T3F442a adipocytes) upon treatment with 10 μM isoproterenol (ISO) or control (CTRL) for 3 hr. (B) Angptl4 mRNA in differentiated brown adipocytes (primary adipocytes and T37i adipocytes) treated with 10 μM isoproterenol (ISO) orcontrol (CTRL) for 3 hr. (C) Immunoblot for ANGPTL4 and LPL protein in differentiated 3T3F422a cells treated with 10 μM isoproterenol (ISO) or control (CTRL) for 3 hr. (D) Lpl mRNA in differentiated mouse primary white or brown adipocytes upon treatment with 10 μM isoproterenol (ISO) or control (CTRL) for 3 hr. (E) Immunoblot for AMPKα1,2 and phospho-AMPK Thr172, AMPKα1, AMPKα2, AMPKβ1 and AMPKβ2 in tissue lysates of kidney, spleen, heart, muscle, liver, inguinal WAT, gonodal WAT and BAT. Homogenates are identical to the homogenates presented in Figure 1A. (F) Immunoblot for AMPKα1,2 and phospho-AMPK Thr172 in BAT and WAT lysates of wild-type mice exposed to 4°C or 28°C for 10 days. *Statistically significant compared to control samples or between indicated treatments according to Student’s t-test (p<0.05). Error bars represent ± SEM. DOI: http://dx.doi.org/10.7554/eLife.08428.012
Figure 7. AMPK is activated in BAT, but not WAT, upon sustained cold exposure. (A) Angptl4 mRNA in differentiated mouse white adipocytes (primary adipocytes and 3T3F442a adipocytes) upon treatment with 10 μM isoproterenol (ISO) or control (CTRL) for 3 hr. (B) Angptl4 mRNA in differentiated brown adipocytes (primary adipocytes and T37i adipocytes) treated with 10 μM isoproterenol (ISO) orcontrol (CTRL) for 3 hr. (C) Immunoblot for ANGPTL4 and LPL protein in differentiated 3T3F422a cells treated with 10 μM isoproterenol (ISO) or control (CTRL) for 3 hr. (D) Lpl mRNA in differentiated mouse primary white or brown adipocytes upon treatment with 10 μM isoproterenol (ISO) or control (CTRL) for 3 hr. (E) Immunoblot for AMPKα1,2 and phospho-AMPK Thr172, AMPKα1, AMPKα2, AMPKβ1 and AMPKβ2 in tissue lysates of kidney, spleen, heart, muscle, liver, inguinal WAT, gonodal WAT and BAT. Homogenates are identical to the homogenates presented in Figure 1A. (F) Immunoblot for AMPKα1,2 and phospho-AMPK Thr172 in BAT and WAT lysates of wild-type mice exposed to 4°C or 28°C for 10 days. *Statistically significant compared to control samples or between indicated treatments according to Student’s t-test (p<0.05). Error bars represent ± SEM. DOI: http://dx.doi.org/10.7554/eLife.08428.012CC BY · PubMed Central
Figure 8. Activation of AMPK down-regulates ANGPTL4 expression specifically in brown adipocytes. (A) Angptl4 mRNA in differentiated T37i adipocytes treated for 6 hr with 1 mM AICAR, 100 μM A769662, 1 mM metformin or 250 μM phenformin hydrochloride. (B) Angptl4 mRNA in differentiated primary brown adipocytes, BA adipocytes, or T37i adipocytes treated for indicated times with 1 mM AICAR. (C) Immunoblot for ANGPTL4, LPL, AMPKα1,2 and phospho-AMPK Thr172 in differentiated T37i cells treated with control (CTRL) or 1 mM AICAR for 3 hr. (D) Angptl4 mRNA in differentiated T37i adipocytes treated with CTRL siRNA or siRNA against AMPKα1 and AMPKα2 for 48 hr, followed by incubation with control (CTRL) or 1 mM AICAR for 3 hr. (E) Angptl4 mRNA in differentiated T37i adipocytes treated with CTRL siRNA or siRNA against AMPKα1 and AMPKα2 for 48 hr, followed by incubation with H2O control medium (CTRL) or 100 μM A769662 for 6 hr. (E) Ampkα1 and Ampkα2 mRNA in differentiated T37i adipocytes treated with CTRL siRNA or siRNA against AMPKα1 and AMPKα2 for 48 hr. (F) Angptl4 mRNA levels in BAT and WAT explants from C57BL/6J wild-type mice (∼50 μg) treated with H2O control medium (CTRL) or 1 mM AICAR for 3 hr. (G) Immunoblot for ANGPTL4 in BAT and WAT explants from C57BL/6J wild-type mice (∼50 mg) treated with H2O control medium (CTRL) or 1 mM AICAR for 3 hr. *Statistically significant compared to control samples or between indicated treatments, according to Student’s t-test (p<0.05). Error bars represent ± SEM. DOI: http://dx.doi.org/10.7554/eLife.08428.013
Figure 8. Activation of AMPK down-regulates ANGPTL4 expression specifically in brown adipocytes. (A) Angptl4 mRNA in differentiated T37i adipocytes treated for 6 hr with 1 mM AICAR, 100 μM A769662, 1 mM metformin or 250 μM phenformin hydrochloride. (B) Angptl4 mRNA in differentiated primary brown adipocytes, BA adipocytes, or T37i adipocytes treated for indicated times with 1 mM AICAR. (C) Immunoblot for ANGPTL4, LPL, AMPKα1,2 and phospho-AMPK Thr172 in differentiated T37i cells treated with control (CTRL) or 1 mM AICAR for 3 hr. (D) Angptl4 mRNA in differentiated T37i adipocytes treated with CTRL siRNA or siRNA against AMPKα1 and AMPKα2 for 48 hr, followed by incubation with control (CTRL) or 1 mM AICAR for 3 hr. (E) Angptl4 mRNA in differentiated T37i adipocytes treated with CTRL siRNA or siRNA against AMPKα1 and AMPKα2 for 48 hr, followed by incubation with H2O control medium (CTRL) or 100 μM A769662 for 6 hr. (E) Ampkα1 and Ampkα2 mRNA in differentiated T37i adipocytes treated with CTRL siRNA or siRNA against AMPKα1 and AMPKα2 for 48 hr. (F) Angptl4 mRNA levels in BAT and WAT explants from C57BL/6J wild-type mice (∼50 μg) treated with H2O control medium (CTRL) or 1 mM AICAR for 3 hr. (G) Immunoblot for ANGPTL4 in BAT and WAT explants from C57BL/6J wild-type mice (∼50 mg) treated with H2O control medium (CTRL) or 1 mM AICAR for 3 hr. *Statistically significant compared to control samples or between indicated treatments, according to Student’s t-test (p<0.05). Error bars represent ± SEM. DOI: http://dx.doi.org/10.7554/eLife.08428.013CC BY · PubMed Central
Figure 9. Down-regulation of Angptl4 expression by AMPK is likely mediated via inhibition of PPARγ-mediated transcription of Angptl4. (A) Angptl4 mRNA in differentiated T37i adipocytes pre-incubated with 0.5 μg/mL actinomycin D (ActD) or DMSO control for 1 hr and treated with 1 mM AICAR or control for 3 hr. (B) Angptl4 mRNA in differentiated T37i or BA adipocytes treated with DMSO control, 5 μM rosiglitazone, 10 μM Wy14643, or 5 μM GW742 (BA adipocytes) or 5 μM GW501516 (T37i adipocytes) for 6 hr. (C) Immunoblot for ANGPTL4 in differentiated T37i adipocytes treated with 5 μM rosiglitazone (Rosi), 10 μM Wy14643 (Wy), and 5 μM GW501516 for 24 hr. (D) Angptl4 mRNA in differentiated T37i cells treated with DMSO control, 5 μM rosiglitazone (Rosi), 1 mM AICAR or both rosiglitazone and AICAR for 6 hr. (E) Relative luciferase activity of HepG2 cells transfected with pGL3-Angptl4, pSG5-Pparγ, pSG5-Rxr, pcDNA-P300HA vectors, as indicated and treated 16 hr post-transfection with 5 μM rosiglitazone, 1 mM AICAR or both compounds for 9 hr. Data are represented as ± SD. (F) Immunoblot of P300 and phospho-P300 (Ser-89) in differentiated T37i brown adipocytes treated with control or 1 mM AICAR for 3 hr. *Statistically significant compared to control samples or between indicated samples according to Student’s t-test (p<0.05). Error bars represent ± SEM, unless otherwise specified. DOI: http://dx.doi.org/10.7554/eLife.08428.014
Figure 9. Down-regulation of Angptl4 expression by AMPK is likely mediated via inhibition of PPARγ-mediated transcription of Angptl4. (A) Angptl4 mRNA in differentiated T37i adipocytes pre-incubated with 0.5 μg/mL actinomycin D (ActD) or DMSO control for 1 hr and treated with 1 mM AICAR or control for 3 hr. (B) Angptl4 mRNA in differentiated T37i or BA adipocytes treated with DMSO control, 5 μM rosiglitazone, 10 μM Wy14643, or 5 μM GW742 (BA adipocytes) or 5 μM GW501516 (T37i adipocytes) for 6 hr. (C) Immunoblot for ANGPTL4 in differentiated T37i adipocytes treated with 5 μM rosiglitazone (Rosi), 10 μM Wy14643 (Wy), and 5 μM GW501516 for 24 hr. (D) Angptl4 mRNA in differentiated T37i cells treated with DMSO control, 5 μM rosiglitazone (Rosi), 1 mM AICAR or both rosiglitazone and AICAR for 6 hr. (E) Relative luciferase activity of HepG2 cells transfected with pGL3-Angptl4, pSG5-Pparγ, pSG5-Rxr, pcDNA-P300HA vectors, as indicated and treated 16 hr post-transfection with 5 μM rosiglitazone, 1 mM AICAR or both compounds for 9 hr. Data are represented as ± SD. (F) Immunoblot of P300 and phospho-P300 (Ser-89) in differentiated T37i brown adipocytes treated with control or 1 mM AICAR for 3 hr. *Statistically significant compared to control samples or between indicated samples according to Student’s t-test (p<0.05). Error bars represent ± SEM, unless otherwise specified. DOI: http://dx.doi.org/10.7554/eLife.08428.014CC BY · PubMed Central

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Research works printed but not linked: 53

  1. Alex, 2013, Short-chain fatty acids stimulate angiopoietin-like 4 synthesis in human colon adenocarcinoma cells by activating peroxisome proliferator-activated receptor, Molecular and Cellular Biology, 33, 1303, 10.1128/MCB.00858-12 · DOI
  2. Alex, 2014, ANGPTL4 is produced by entero-endocrine cells in the human intestinal tract, Histochemistry and Cell Biology, 141, 383, 10.1007/s00418-013-1157-y · DOI · Paper page
  3. An, 2005, beta-Agonist stimulation produces changes in cardiac AMPK and coronary lumen LPL only during increased workload, AJP: Endocrinology and Metabolism, 288, 10.1152/ajpendo.00588.2004 · DOI
  4. Bartelt, 2013, Effects of adipocyte lipoprotein lipase on de novo lipogenesis and white adipose tissue browning, Biochimica Et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, 1831, 934, 10.1016/j.bbalip.2012.11.011 · DOI
  5. Bauwens, 2011, Cold tolerance, cold-induced hyperphagia, and nonshivering thermogenesis are normal in α1-AMPK-/- mice, AJP: Regulatory, Integrative and Comparative Physiology, 301, R473, 10.1152/ajpregu.00444.2010 · DOI
  6. Berbée, 2015, Brown fat activation reduces hypercholesterolaemia and protects from atherosclerosis development, Nature Communications, 6:, 10.1038/ncomms7356 · DOI
  7. Bergeron, 2001, Effect of 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside infusion on in vivo glucose and lipid metabolism in lean and obese zucker rats, Diabetes, 50, 1076, 10.2337/diabetes.50.5.1076 · DOI
  8. Bertin, 1985, Effects of cold acclimation on the activity of lipoprotein lipase in adipose tissues of genetically obese zucker rats, Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 81, 797, 10.1016/0305-0491(85)90408-0 · DOI
  9. Bruns, 2009, Real-time magnetic resonance imaging and quantification of lipoprotein metabolism in vivo using nanocrystals, Nature Nanotechnology, 4, 193, 10.1038/nnano.2008.405 · DOI
  10. Buhl, 2002, Long-term AICAR administration reduces metabolic disturbances and lowers blood pressure in rats displaying features of the insulin resistance syndrome, Diabetes, 51, 2199, 10.2337/diabetes.51.7.2199 · DOI
  11. Carneheim, 1988, Cold-induced beta-adrenergic recruitment of lipoprotein lipase in brown fat is due to increased transcription, The American Journal of Physiology, 254
  12. Catoire, 2014, Fatty acid-inducible ANGPTL4 governs lipid metabolic response to exercise, Proceedings of the National Academy of Sciences of the United States of America, 111, 10.1073/pnas.1400889111 · DOI · Paper page
  13. Cheang, 2014, Metformin protects endothelial function in diet-induced obese mice by inhibition of endoplasmic reticulum stress through 5' adenosine monophosphate-activated protein kinase-peroxisome proliferator-activated receptor pathway, Arteriosclerosis, Thrombosis, and Vascular Biology, 34, 830, 10.1161/ATVBAHA.113.301938 · DOI
  14. Dijk, 2014, Regulation of lipoprotein lipase by Angptl4, Trends in Endocrinology & Metabolism, 25, 146, 10.1016/j.tem.2013.12.005 · DOI
  15. Garcia-Arcos, 2013, Adipose-specific lipoprotein lipase deficiency more profoundly affects brown than white fat biology, Journal of Biological Chemistry, 288, 14046, 10.1074/jbc.M113.469270 · DOI · Paper page
  16. Ge, 2004, Oligomerization state-dependent hyperlipidemic effect of angiopoietin-like protein 4, The Journal of Lipid Research, 45, 2071, 10.1194/jlr.M400138-JLR200 · DOI · Paper page
  17. Geerling, 2014, Metformin lowers plasma triglycerides by promoting VLDL-triglyceride clearance by brown adipose tissue in mice, Diabetes, 63, 880, 10.2337/db13-0194 · DOI
  18. Gelman, 1999, P300 interacts with the n- and c-terminal part of PPAR 2 in a ligand-independent and -dependent manner, respectively, Journal of Biological Chemistry, 274, 7681, 10.1074/jbc.274.12.7681 · DOI
  19. Georgiadi, 2010, Induction of cardiac Angptl4 by dietary fatty acids is mediated by peroxisome proliferator-activated receptor beta /delta and protects against fatty acid-induced oxidative stress, Circulation Research, 106, 1712, 10.1161/CIRCRESAHA.110.217380 · DOI · Paper page
  20. Giralt, 1990, Lipoprotein lipase mRNA expression in brown adipose tissue: translational and/or posttranslational events are involved in the modulation of enzyme activity, Biochimica Et Biophysica Acta (BBA) - Gene Structure and Expression, 1048, 270, 10.1016/0167-4781(90)90066-B · DOI
  21. Jong, 2001, Apolipoprotein c-III deficiency accelerates triglyceride hydrolysis by lipoprotein lipase in wild-type and apoE knockout mice, Journal of Lipid Research, 42, 1578, 10.1016/S0022-2275(20)32211-2 · DOI
  22. Kaddatz, 2010, Transcriptional profiling identifies functional interactions of TGFβ and PPARβ/δ signaling, Journal of Biological Chemistry, 285, 29469, 10.1074/jbc.M110.142018 · DOI
  23. Kersten, 2000, Characterization of the fasting-induced adipose factor FIAF, a novel peroxisome proliferator-activated receptor target gene, Journal of Biological Chemistry, 275, 28488, 10.1074/jbc.M004029200 · DOI · Paper page
  24. Kersten, 2009, Caloric restriction and exercise increase plasma ANGPTL4 levels in humans via elevated free fatty acids, Arteriosclerosis, Thrombosis, and Vascular Biology, 29, 969, 10.1161/ATVBAHA.108.182147 · DOI · Paper page
  25. Kersten, 2014, Physiological regulation of lipoprotein lipase, Biochimica Et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, 1841, 919, 10.1016/j.bbalip.2014.03.013 · DOI
  26. Khedoe, 2014, Brown adipose tissue takes up plasma triglycerides mostly after lipolysis, Journal of Lipid Research, 569, 51
  27. Kim, 2000, Hepatic expression, synthesis and secretion of a novel fibrinogen/angiopoietin-related protein that prevents endothelial-cell apoptosis, Biochemical Journal, 346, 603, 10.1042/bj3460603 · DOI
  28. Klingenspor, 1989, Short photoperiod and cold activate brown fat lipoprotein lipase in the djungarian hamster, The American Journal of Physiology, 257, R1123
  29. Klingenspor, 1996, Multiple regulatory steps are involved in the control of lipoprotein lipase activity in brown adipose tissue, Journal of Lipid Research, 37, 1685, 10.1016/S0022-2275(20)39111-2 · DOI
  30. Kroupa, 2012, Linking nutritional regulation of Angptl4, Gpihbp1, and Lmf1 to lipoprotein lipase activity in rodent adipose tissue, BMC Physiology, 12, 13, 10.1159/000327774 · DOI
  31. Leff, 2003, AMP-activated protein kinase regulates gene expression by direct phosphorylation of nuclear proteins, Biochemical Society Transactions, 31, 224, 10.1042/bst0310224 · DOI
  32. Lichtenstein, 2010, Angptl4 protects against severe proinflammatory effects of saturated fat by inhibiting fatty acid uptake into mesenteric lymph node macrophages, Cell Metabolism, 12, 580, 10.1016/j.cmet.2010.11.002 · DOI · Paper page
  33. Lim, 2012, AMP-activated protein kinase inhibits TGF-β-induced fibrogenic responses of hepatic stellate cells by targeting transcriptional coactivator p300, Journal of Cellular Physiology, 227, 1081, 10.1002/jcp.22824 · DOI
  34. Mandard, 2004, The direct peroxisome proliferator-activated receptor target fasting-induced adipose factor (FIAF/PGAR/ANGPTL4) is present in blood plasma as a truncated protein that is increased by fenofibrate treatment, Journal of Biological Chemistry, 279, 34411, 10.1074/jbc.M403058200 · DOI · Paper page
  35. Mandard, 2006, The fasting-induced adipose Factor/Angiopoietin-like protein 4 is physically associated with lipoproteins and governs plasma lipid levels and adiposity, Journal of Biological Chemistry, 281, 934, 10.1074/jbc.M506519200 · DOI · Paper page
  36. Mitchell, 1992, Regulation of expression of the lipoprotein lipase gene in brown adipose tissue, The American Journal of Physiology, 263
  37. Mulligan, 2007, Upregulation of AMPK during cold exposure occurs via distinct mechanisms in brown and white adipose tissue of the mouse, The Journal of Physiology, 580, 677, 10.1113/jphysiol.2007.128652 · DOI
  38. Namgaladze, 2013, AICAR inhibits PPAR during monocyte differentiation to attenuate inflammatory responses to atherogenic lipids, Cardiovascular Research, 98, 479, 10.1093/cvr/cvt073 · DOI
  39. Ohira, 2009, Metformin promotes induction of lipoprotein lipase in skeletal muscle through activation of adenosine monophosphate-activated protein kinase, Metabolism, 58, 1408, 10.1016/j.metabol.2009.04.024 · DOI
  40. Rensen, 1995, Selective liver targeting of antivirals by recombinant chylomicrons—a new therapeutic approach to hepatitis b, Nature Medicine, 1, 221, 10.1038/nm0395-221 · DOI
  41. Ruge, 2004, Nutritional regulation of lipoprotein lipase in mice, The International Journal of Biochemistry & Cell Biology, 36, 320, 10.1016/S1357-2725(03)00256-5 · DOI
  42. Sozio, 2011, Activated AMPK inhibits PPAR- and PPAR- transcriptional activity in hepatoma cells, AJP: Gastrointestinal and Liver Physiology, 301, G739, 10.1152/ajpgi.00432.2010 · DOI
  43. Uldry, 2006, Complementary action of the PGC-1 coactivators in mitochondrial biogenesis and brown fat differentiation, Cell Metabolism, 3, 333, 10.1016/j.cmet.2006.04.002 · DOI
  44. Vijgen, 2013, Increased oxygen consumption in human adipose tissue from the 'brown adipose tissue' region, The Journal of Clinical Endocrinology & Metabolism, 98, 10.1210/jc.2013-1348 · DOI
  45. Vo, 2001, CREB-binding protein and p300 in transcriptional regulation, Journal of Biological Chemistry, 276, 13505, 10.1074/jbc.R000025200 · DOI
  46. Wang, 2011, Brown adipose tissue in humans is activated by elevated plasma catecholamines levels and is inversely related to central obesity, PLoS ONE, 6, 10.1371/journal.pone.0021006.t002 · DOI
  47. Wijers, 2010, Cold-induced adaptive thermogenesis in lean and obese, Obesity, 18, 1092, 10.1038/oby.2010.74 · DOI
  48. Wu, 2012, Beige adipocytes are a distinct type of thermogenic fat cell in mouse and human, Cell, 150, 366, 10.1016/j.cell.2012.05.016 · DOI
  49. Yang, 2001, Regulation of transcription by AMP-activated protein kinase: PHOSPHORYLATION of p300 BLOCKS ITS INTERACTION with NUCLEAR RECEPTORS, Journal of Biological Chemistry, 276, 38341, 10.1074/jbc.C100316200 · DOI
  50. Yang, 2008, Suppression of the Raf/MEK/ERK signaling cascade and inhibition of angiogenesis by the carboxyl terminus of angiopoietin-like protein 4, Arteriosclerosis, Thrombosis, and Vascular Biology, 28, 835, 10.1161/ATVBAHA.107.157776 · DOI
  51. Yoon, 2000, Peroxisome proliferator-activated receptor gamma target gene encoding a novel angiopoietin-related protein associated with adipose differentiation, Molecular and Cellular Biology, 20, 5343, 10.1128/MCB.20.14.5343-5349.2000 · DOI · Paper page
  52. Young, 2013, Biochemistry and pathophysiology of intravascular and intracellular lipolysis, Genes & Development, 27, 459, 10.1101/gad.209296.112 · DOI
  53. Zennaro, 1998, Hibernoma development in transgenic mice identifies brown adipose tissue as a novel target of aldosterone action, Journal of Clinical Investigation, 101, 1254, 10.1172/JCI1915 · DOI

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Authors
Dijk, W., Heine, M., Vergnes, L., Boon, M. R., Schaart, G., Hesselink, M. K., Reue, K., van Marken Lichtenbelt, W. D., Olivecrona, G., Rensen, P. C., Heeren, J., & Kersten, S.
Affiliations
  • Nutrition, Metabolism and Genomics group, Division of Human Nutrition, Wageningen University, Wageningen, The Netherlands
  • Department of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
  • Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, United States
  • Department of Medicine, Division of Endocrinology, Leiden University Medical Center, Leiden, The Netherlands
  • Department of Human Movement Sciences, Maastricht University, Maastricht, The Netherlands
  • Department of Human Biology, Maastricht University Medical Centre, Maastricht, The Netherlands
  • Department of Medical Biosciences/Physiological Chemistry, Umeå University, Umeå, Sweden
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2015-10-17
Volume
4
ISSN
2050-084X
DOI
10.7554/elife.08428
PMC
PMC4709329
Cited by
114
References
8
Language
en
Licence
Funders
  • Fondation Leducq (12CVD04)
  • Forskningsrådet om Hälsa, Arbetsliv och Välfärd (12203)
  • Hartstichting (2009T038)

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