Carnitine Palmitoyltransferase 1 Increases Lipolysis, UCP1 Protein Expression and Mitochondrial Activity in Brown Adipocytes

Calderón‐Domínguez, María; Sebastián, David; Fucho, Raquel; Weber, Minéia; Mir, Joan Francesc; García-Casarrubios, Ester; Obregón, Marı́a Jesús; Zorzano, António; Valverde, Ángela M.; Serra, Dolors; Herrero, Laura

doi:10.1371/journal.pone.0159399

Cited by 56 publications

(36 citation statements)

References 55 publications

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“…The adipocytes depended heavily on FA oxidation when stimulated with isoproterenol. Numerous studies support the notion that modulation of the mitochondrial FA carriers affects FA oxidation and thermogenesis in brown adipocytes (7,8,21,25), underlining the important role of FAs not only as substrates but also as activators of thermogenesis (9). Respiration also depended heavily on glucose oxidation, although the glucose pathway flexibility was very low in ISO-treated cells.…”

Section: Discussionmentioning

confidence: 79%

Restricting glycolysis impairs brown adipocyte glucose and oxygen consumption

Winther

Isidor

Basse

et al. 2018

American Journal of Physiology-Endocrinology and Metabolism

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During thermogenic activation, brown adipocytes take up large amounts of glucose. In addition, cold stimulation leads to an upregulation of glycolytic enzymes. Here we have investigated the importance of glycolysis for brown adipocyte glucose consumption and thermogenesis. Using siRNA-mediated knockdown in mature adipocytes, we explored the effect of glucose transporters and glycolytic enzymes on brown adipocyte functions such as consumption of glucose and oxygen. Basal oxygen consumption in brown adipocytes was equally dependent on glucose and fatty acid oxidation, whereas isoproterenol (ISO)-stimulated respiration was fueled mainly by fatty acids, with a significant contribution from glucose oxidation. Knockdown of glucose transporters in brown adipocytes not only impaired ISO-stimulated glycolytic flux but also oxygen consumption. Diminishing glycolytic flux by knockdown of the first and final enzyme of glycolysis, i.e., hexokinase 2 (HK2) and pyruvate kinase M (PKM), respectively, decreased glucose uptake and ISO-stimulated oxygen consumption. HK2 knockdown had a more severe effect, which, in contrast to PKM knockdown, could not be rescued by supplementation with pyruvate. Hence, brown adipocytes rely on glucose consumption and glycolytic flux to achieve maximum thermogenic output, with glycolysis likely supporting thermogenesis not only by pyruvate formation but also by supplying intermediates for efferent metabolic pathways.

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Section: Discussionmentioning

confidence: 79%

Restricting glycolysis impairs brown adipocyte glucose and oxygen consumption

Winther

Isidor

Basse

et al. 2018

American Journal of Physiology-Endocrinology and Metabolism

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“…fetal serum, 1% (vol./vol.) penicillin/streptomycin (Sigma-Aldrich) and 2 mmol/l HEPES, and differentiated until they exhibited numerous multilocular cytoplasmic lipid droplets [26]. Undifferentiated (day 0) pre-adipocytes were treated with 0.1 μg/ml PTN (Sigma-Aldrich) until day 6 of differentiation; at this time point, non-treated pre-adipocytes typically achieved a fully differentiated phenotype.…”

Section: Cell Culture and Differentiation Of Brown Adipocytesmentioning

confidence: 99%

Pleiotrophin deletion alters glucose homeostasis, energy metabolism and brown fat thermogenic function in mice

et al. 2018

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Aims/hypothesis Pleiotrophin, a developmentally regulated and highly conserved cytokine, exerts different functions including regulation of cell growth and survival. Here, we hypothesise that this cytokine can play a regulatory role in glucose and lipid homeostasis. Methods To test this hypothesis, we performed a longitudinal study characterising the metabolic profile (circulating variables and tissue mRNA expression) of gene-targeted Ptn-deficient female mice and their corresponding wild-type counterparts at different ages from young adulthood (3 months) to older age (15 months). Metabolic cages were used to investigate the respiratory exchange ratio and energy expenditure, at both 24°C and 30°C. Undifferentiated immortalised mouse brown adipocytes (mBAs) were treated with 0.1 μg/ml pleiotrophin until day 6 of differentiation, and markers of mBA differentiation were analysed by quantitative real-time PCR (qPCR). Results Ptn deletion was associated with a reduction in total body fat (20.2% in Ptn +/+ vs 13.9% in Ptn −/− mice) and an enhanced lipolytic response to isoprenaline in isolated adipocytes from 15-month-old mice (189% in Ptn +/+ vs 273% in Ptn −/− mice). We found that Ptn −/− mice exhibited a significantly lower QUICKI value and an altered lipid profile; plasma triacylglycerols and NEFA did not increase with age, as happens in Ptn +/+ mice. Furthermore, the contribution of cold-induced thermogenesis to energy expenditure was greater in Ptn −/− than Ptn +/+ mice (42.6% and 33.6%, respectively). Body temperature and the activity and expression of deiodinase, T 3 and mitochondrial uncoupling protein-1 in the brown adipose tissue of Ptn −/− mice were higher than in wild-type controls. Finally, supplementing brown pre-adipocytes with pleiotrophin decreased the expression of the brown adipocyte markers Cidea (20% reduction), Prdm16 (21% reduction), and Pgc1-α (also known as Ppargc1a, 11% reduction). Conclusions/interpretation Our results reveal for the first time that pleiotrophin is a key player in preserving insulin sensitivity, driving the dynamics of adipose tissue lipid turnover and plasticity, and regulating energy metabolism and thermogenesis. These findings open therapeutic avenues for the treatment of metabolic disorders by targeting pleiotrophin in the crosstalk between white and brown adipose tissue.

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“…According to all these algorithms, our results show that the three most stable genes were Hprt1, Tbp, and Rpl32 for IBAT and Tbp, B2m, and Hprt1 for RWAT, while the least stable genes were 18S, Actb, and Gapdh for both tissues. The Actb and Gapdh genes are widely used in the literature as normalizers for the evaluation of target gene expression in WAT and BAT (Aguirre et al, 2016;Antony et al, 2017;Calderon-Dominguez et al, 2016;Chang and Kim, 2017;Li et al, 2015;Rodrigues et al, 2017;Tsubai et al, 2016;Yan et al, 2017), however, a few authors argue about the stability of the reference genes. A systematic review showed that the vast majority of the studies use a single gene as a normalizer, with Actb and/or Gapdh being commonly selected to evaluate the gene expression in vertebrates (Chapman and Waldenstrom, 2015).…”

Section: Discussionmentioning

confidence: 99%

Identification of Suitable Reference Genes for Quantitative Gene Expression Analysis in Innervated and Denervated Adipose Tissue from Cafeteria Diet‐Fed Rats

Marques-Oliveira

Silva

Valadares

et al. 2019

Lipids

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Our previous studies show that cafeteria diet increases body adiposity, plasma insulin levels, and sympathetic activity to brown adipose tissue (BAT) and white adipose tissue (WAT) of Wistar rats, leading to rapid and progressive changes in the metabolic profile. The identification of suitable reference genes that are not affected by the experimental conditions is a critical step in accurate normalization of the reverse transcription quantitative real‐time PCR (qRT‐PCR), a commonly used assay to elucidate changes in the gene expression profile. In the present study, the effects of the cafeteria diet and sympathetic innervation on the gene expression of adrenoceptor beta 3 (Adrb3) from BAT and WAT were assessed using one of the most stable and one of the least stable genes as normalizers. Rats were fed the cafeteria diet and on the 17th day, interscapular BAT or retroperitoneal WAT was denervated and, 7 days after surgery, the contralateral innervated tissue was used as control. Ten reference genes were evaluated (18S, B2m, Actb, CypA, Gapdh, Hprt1, Rpl32, Tbp, Ubc, and Ywhaz) and ranked according to their stability using the following algorithms: geNorm, NormFinder, BestKeeper, and comparative delta threshold cycle (ΔC t) method. According to the algorithms employed, the normalization of Adrb3 expression by the least stable genes produced opposite results compared with the most stable genes and literature data. In cafeteria and control diet‐fed rats, the three most stable genes were Hprt1, Tbp, and Rpl32 for interscapular BAT and Tbp, B2m, and Hprt1 for retroperitoneal WAT, while the least stable genes were 18S, Actb, and Gapdh for both tissues.

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Carnitine Palmitoyltransferase 1 Increases Lipolysis, UCP1 Protein Expression and Mitochondrial Activity in Brown Adipocytes

Cited by 56 publications

References 55 publications

Restricting glycolysis impairs brown adipocyte glucose and oxygen consumption

Restricting glycolysis impairs brown adipocyte glucose and oxygen consumption

Pleiotrophin deletion alters glucose homeostasis, energy metabolism and brown fat thermogenic function in mice

Identification of Suitable Reference Genes for Quantitative Gene Expression Analysis in Innervated and Denervated Adipose Tissue from Cafeteria Diet‐Fed Rats

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