36 results on '"Zechner, Rudolf"'
Search Results
2. Lipolysis meets inflammation: arachidonic acid mobilization from fat1
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Schreiber, Renate and Zechner, Rudolf
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Arachidonic Acid ,Prostaglandin D2 ,Lipolysis ,Phospholipases A2, Cytosolic ,Antigens, CD34 ,Lipase ,Lipid Droplets ,QD415-436 ,Immunoglobulin E ,Biochemistry ,Leukotriene C4 ,Kinetics ,Cyclooxygenase 2 ,Commentary ,Cyclooxygenase 1 ,Eicosanoids ,Humans ,Gene Silencing ,Mast Cells ,RNA, Small Interfering ,Cells, Cultured ,Triglycerides - Abstract
Human mast cells (MCs) contain TG-rich cytoplasmic lipid droplets (LDs) with high arachidonic acid (AA) content. Here, we investigated the functional role of adipose TG lipase (ATGL) in TG hydrolysis and the ensuing release of AA as substrate for eicosanoid generation by activated human primary MCs in culture. Silencing of ATGL in MCs by siRNAs induced the accumulation of neutral lipids in LDs. IgE-dependent activation of MCs triggered the secretion of the two major eicosanoids, prostaglandin D2 (PGD2) and leukotriene C4 (LTC4). The immediate release of PGD2 from the activated MCs was solely dependent on cyclooxygenase (COX) 1, while during the delayed phase of lipid mediator production, the inducible COX-2 also contributed to its release. Importantly, when ATGL-silenced MCs were activated, the secretion of both PGD2 and LTC4 was significantly reduced. Interestingly, the inhibitory effect on the release of LTC4 was even more pronounced in ATGL-silenced MCs than in cytosolic phospholipase A2-silenced MCs. These data show that ATGL hydrolyzes AA-containing TGs present in human MC LDs and define ATGL as a novel regulator of the substrate availability of AA for eicosanoid generation upon MC activation.
- Published
- 2014
3. Cold-induced phosphatidylethanolamine synthesis in liver and brown adipose tissue of mice.
- Author
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Hidrobo MS, Höring M, Brunner S, Liebisch G, Schweizer S, Klingenspor M, Schreiber R, Zechner R, Burkhardt R, and Ecker J
- Abstract
Increasing energy expenditure in brown adipose (BAT) tissue by cold-induced lipolysis is discussed as a potential strategy to counteract imbalanced lipid homeostasis caused through unhealthy lifestyle and cardiometabolic disease. Yet, it is largely unclear how liberated fatty acids (FA) are metabolized. We investigated the liver and BAT lipidome of mice housed for 1 week at thermoneutrality, 23 °C and 4 °C using quantitative mass spectrometry-based lipidomics. Housing at temperatures below thermoneutrality triggered the generation of phosphatidylethanolamine (PE) in both tissues. Particularly, the concentrations of PE containing polyunsaturated fatty acids (PUFA) in their acyl chains like PE 18: 0_20:4 were increased at cold. Investigation of the plasma's FA profile using gas chromatography coupled to mass spectrometry revealed a negative correlation of PUFA with unsaturated PE in liver and BAT indicating a flux of FA from the circulation into these tissues. Beta-adrenergic stimulation elevated intracellular levels of PE 38:4 and PE 40:6 in beige wildtype adipocytes, but not in adipose triglyceride lipase (ATGL)-deficient cells. These results imply an induction of PE synthesis in liver, BAT and thermogenic adipocytes after activation of the beta-adrenergic signaling cascade., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)
- Published
- 2024
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4. Correction: Unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification-58.
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Kulminskaya N, Rodriguez Gamez CF, Hofer P, Cerk IK, Dubey N, Viertlmayr R, Sagmeister T, Pavkov-Keller T, Zechner R, and Oberer M
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- 2024
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5. Corrigendum to "PNPLA-mediated lipid hydrolysis and transacylation - At the intersection of catabolism and anabolism" [Biochim. Biophys. Acta (BBA) - Mol. Cell Biol. Lipids volume 1869, issue 2, March 2024, 159,410].
- Author
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Colaço-Gaspar M, Hofer P, Oberer M, and Zechner R
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- 2024
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6. PNPLA-mediated lipid hydrolysis and transacylation - At the intersection of catabolism and anabolism.
- Author
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Colaço-Gaspar M, Hofer P, Oberer M, and Zechner R
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- Hydrolysis, Epidermis metabolism, Lipids, Lipase metabolism, Lipid Metabolism
- Abstract
Patatin-like phospholipase domain containing proteins (PNPLAs) play diverse roles in lipid metabolism. In this review, we focus on the enzymatic properties and predicted 3D structures of PNPLA1-5. PNPLA2-4 exert both catabolic and anabolic functions. Whereas PNPLA1 is predominantly expressed in the epidermis and involved in sphingolipid biosynthesis, PNPLA2 and 4 are ubiquitously expressed and exhibit several enzymatic activities, including hydrolysis and transacylation of various (glycero-)lipid species. This review summarizes known biological roles for PNPLA-mediated hydrolysis and transacylation reactions and highlights open questions concerning their physiological function., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2024
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7. Unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification-58.
- Author
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Kulminskaya N, Rodriguez Gamez CF, Hofer P, Cerk IK, Dubey N, Viertlmayr R, Sagmeister T, Pavkov-Keller T, Zechner R, and Oberer M
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- Animals, Mice, Lipolysis physiology, Triglycerides metabolism, Amino Acids metabolism, 1-Acylglycerol-3-Phosphate O-Acyltransferase metabolism, Artificial Intelligence, Lipase metabolism
- Abstract
Lipolysis is an essential metabolic process that releases unesterified fatty acids from neutral lipid stores to maintain energy homeostasis in living organisms. Adipose triglyceride lipase (ATGL) plays a key role in intracellular lipolysis and can be coactivated upon interaction with the protein comparative gene identification-58 (CGI-58). The underlying molecular mechanism of ATGL stimulation by CGI-58 is incompletely understood. Based on analysis of evolutionary conservation, we used site directed mutagenesis to study a C-terminally truncated variant and full-length mouse ATGL providing insights in the protein coactivation on a per-residue level. We identified the region from residues N209-N215 in ATGL as essential for coactivation by CGI-58. ATGL variants with amino acids exchanges in this region were still able to hydrolyze triacylglycerol at the basal level and to interact with CGI-58, yet could not be activated by CGI-58. Our studies also demonstrate that full-length mouse ATGL showed higher tolerance to specific single amino acid exchanges in the N209-N215 region upon CGI-58 coactivation compared to C-terminally truncated ATGL variants. The region is either directly involved in protein-protein interaction or essential for conformational changes required in the coactivation process. Three-dimensional models of the ATGL/CGI-58 complex with the artificial intelligence software AlphaFold demonstrated that a large surface area is involved in the protein-protein interaction. Mapping important amino acids for coactivation of both proteins, ATGL and CGI-58, onto the 3D model of the complex locates these essential amino acids at the predicted ATGL/CGI-58 interface thus strongly corroborating the significance of these residues in CGI-58-mediated coactivation of ATGL., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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8. Cooperative lipolytic control of neuronal triacylglycerol by spastic paraplegia-associated enzyme DDHD2 and ATGL.
- Author
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Hofer P, Grabner GF, König M, Xie H, Bulfon D, Ludwig AE, Wolinski H, Zimmermann R, Zechner R, and Heier C
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- Humans, Lipase genetics, Lipase metabolism, Neurons metabolism, Paraplegia, Phospholipases metabolism, Triglycerides metabolism, Lipolysis
- Abstract
Intracellular lipolysis-the enzymatic breakdown of lipid droplet-associated triacylglycerol (TAG)-depends on the cooperative action of several hydrolytic enzymes and regulatory proteins, together designated as lipolysome. Adipose triglyceride lipase (ATGL) acts as a major cellular TAG hydrolase and core effector of the lipolysome in many peripheral tissues. Neurons initiate lipolysis independently of ATGL via DDHD domain-containing 2 (DDHD2), a multifunctional lipid hydrolase whose dysfunction causes neuronal TAG deposition and hereditary spastic paraplegia. Whether and how DDHD2 cooperates with other lipolytic enzymes is currently unknown. In this study, we further investigated the enzymatic properties and functions of DDHD2 in neuroblastoma cells and primary neurons. We found that DDHD2 hydrolyzes multiple acylglycerols in vitro and substantially contributes to neutral lipid hydrolase activities of neuroblastoma cells and brain tissue. Substrate promiscuity of DDHD2 allowed its engagement at different steps of the lipolytic cascade: In neuroblastoma cells, DDHD2 functioned exclusively downstream of ATGL in the hydrolysis of sn-1,3-diacylglycerol (DAG) isomers but was dispensable for TAG hydrolysis and lipid droplet homeostasis. In primary cortical neurons, DDHD2 exhibited lipolytic control over both, DAG and TAG, and complemented ATGL-dependent TAG hydrolysis. We conclude that neuronal cells use noncanonical configurations of the lipolysome and engage DDHD2 as dual TAG/DAG hydrolase in cooperation with ATGL., Competing Interests: Conflict of interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2023
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9. Rosiglitazone Reverses Inflammation in Epididymal White Adipose Tissue in Hormone-Sensitive Lipase-Knockout Mice.
- Author
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Kotzbeck P, Taschler U, Haudum C, Foessl I, Schoiswohl G, Boulgaropoulos B, Bounab K, Einsiedler J, Pajed L, Tilp A, Schwarz A, Eichmann TO, Obermayer-Pietsch B, Giordano A, Cinti S, Zechner R, and Pieber TR
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- Mice, Animals, Rosiglitazone pharmacology, Mice, Knockout, Adipose Tissue metabolism, Adipose Tissue, White metabolism, Lipolysis physiology, Inflammation drug therapy, Inflammation genetics, Inflammation metabolism, Sterol Esterase genetics, Sterol Esterase metabolism, PPAR gamma genetics, PPAR gamma metabolism
- Abstract
Hormone-sensitive lipase (HSL) plays a crucial role in intracellular lipolysis, and loss of HSL leads to diacylglycerol (DAG) accumulation, reduced FA mobilization, and impaired PPARγ signaling. Hsl knockout mice exhibit adipose tissue inflammation, but the underlying mechanisms are still not clear. Here, we investigated if and to what extent HSL loss contributes to endoplasmic reticulum (ER) stress and adipose tissue inflammation in Hsl knockout mice. Furthermore, we were interested in how impaired PPARγ signaling affects the development of inflammation in epididymal white adipose tissue (eWAT) and inguinal white adipose tissue (iWAT) of Hsl knockout mice and if DAG and ceramide accumulation contribute to adipose tissue inflammation and ER stress. Ultrastructural analysis showed a markedly dilated ER in both eWAT and iWAT upon loss of HSL. In addition, Hsl knockout mice exhibited macrophage infiltration and increased F4/80 mRNA expression, a marker of macrophage activation, in eWAT, but not in iWAT. We show that treatment with rosiglitazone, a PPARγ agonist, attenuated macrophage infiltration and ameliorated inflammation of eWAT, but expression of ER stress markers remained unchanged, as did DAG and ceramide levels in eWAT. Taken together, we show that HSL loss promoted ER stress in both eWAT and iWAT of Hsl knockout mice, but inflammation and macrophage infiltration occurred mainly in eWAT. Also, PPARγ activation reversed inflammation but not ER stress and DAG accumulation. These data indicate that neither reduction of DAG levels nor ER stress contribute to the reversal of eWAT inflammation in Hsl knockout mice., Competing Interests: Conflict of Interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2023
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10. Adipose triglyceride lipase activity regulates cancer cell proliferation via AMP-kinase and mTOR signaling.
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Xie H, Heier C, Kien B, Vesely PW, Tang Z, Sexl V, Schoiswohl G, Strießnig-Bina I, Hoefler G, Zechner R, and Schweiger M
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- Animals, Cell Line, Fibroblasts metabolism, Humans, Lipolysis, Mice, Signal Transduction, Adenylate Kinase metabolism, Cell Proliferation, Lipase metabolism, Neoplasms metabolism, Neoplasms pathology, TOR Serine-Threonine Kinases metabolism
- Abstract
Aberrant fatty acid (FA) metabolism is a hallmark of proliferating cells, including untransformed fibroblasts or cancer cells. Lipolysis of intracellular triglyceride (TG) stores by adipose triglyceride lipase (ATGL) provides an important source of FAs serving as energy substrates, signaling molecules, and precursors for membrane lipids. To investigate if ATGL-mediated lipolysis impacts cell proliferation, we modified ATGL activity in murine embryonic fibroblasts (MEFs) and in five different cancer cell lines to determine the consequences on cell growth and metabolism. Genetic or pharmacological inhibition of ATGL in MEFs causes impaired FA oxidation, decreased ROS production, and a substrate switch from FA to glucose leading to decreased AMPK-mTOR signaling and higher cell proliferation rates. ATGL expression in these cancer cells is low when compared to MEFs. Additional ATGL knockdown in cancer cells did not significantly affect cellular lipid metabolism or cell proliferation whereas the ectopic overexpression of ATGL increased lipolysis and reduced proliferation. In contrast to ATGL silencing, pharmacological inhibition of ATGL by Atglistatin© impeded the proliferation of diverse cancer cell lines, which points at an ATGL-independent effect. Our data indicate a crucial role of ATGL-mediated lipolysis in the regulation of cell proliferation. The observed low ATGL activity in cancer cells may represent an evolutionary selection process and mechanism to sustain high cell proliferation rates. As the increasing ATGL activity decelerates proliferation of five different cancer cell lines this may represent a novel therapeutic strategy to counteract uncontrolled cell growth., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2020
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11. Metabolic disease and ABHD6 alter the circulating bis(monoacylglycerol)phosphate profile in mice and humans.
- Author
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Grabner GF, Fawzy N, Pribasnig MA, Trieb M, Taschler U, Holzer M, Schweiger M, Wolinski H, Kolb D, Horvath A, Breinbauer R, Rülicke T, Rabl R, Lass A, Stadlbauer V, Hutter-Paier B, Stauber RE, Fickert P, Zechner R, Marsche G, Eichmann TO, and Zimmermann R
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- Adult, Aged, Animals, Female, Humans, Lysophospholipids blood, Male, Metabolic Diseases blood, Mice, Mice, Knockout, Mice, Transgenic, Middle Aged, Monoacylglycerol Lipases deficiency, Monoacylglycerol Lipases genetics, Monoglycerides blood, Phenotype, Lysophospholipids metabolism, Metabolic Diseases metabolism, Monoacylglycerol Lipases metabolism, Monoglycerides metabolism
- Abstract
Bis(monoacylglycerol)phosphate (BMP) is a phospholipid that is crucial for lipid degradation and sorting in acidic organelles. Genetic and drug-induced lysosomal storage disorders (LSDs) are associated with increased BMP concentrations in tissues and in the circulation. Data on BMP in disorders other than LSDs, however, are scarce, and key enzymes regulating BMP metabolism remain elusive. Here, we demonstrate that common metabolic disorders and the intracellular BMP hydrolase α/β-hydrolase domain-containing 6 (ABHD6) affect BMP metabolism in mice and humans. In mice, dietary lipid overload strongly affects BMP concentration and FA composition in the liver and plasma, similar to what has been observed in LSDs. Notably, distinct changes in the BMP FA profile enable a clear distinction between lipid overload and drug-induced LSDs. Global deletion of ABHD6 increases circulating BMP concentrations but does not cause LSDs. In humans, nonalcoholic fatty liver disease and liver cirrhosis affect the serum BMP FA composition and concentration. Furthermore, we identified a patient with a loss-of-function mutation in the ABHD6 gene, leading to an altered circulating BMP profile. In conclusion, our results suggest that common metabolic diseases and ABHD6 affect BMP metabolism in mice and humans., (Copyright © 2019 Grabner et al. Published by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2019
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12. Brown adipose tissue whitening leads to brown adipocyte death and adipose tissue inflammation.
- Author
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Kotzbeck P, Giordano A, Mondini E, Murano I, Severi I, Venema W, Cecchini MP, Kershaw EE, Barbatelli G, Haemmerle G, Zechner R, and Cinti S
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- Animals, Lipase deficiency, Lipase metabolism, Mice, Mice, Inbred C57BL, Mice, Knockout, Adipose Tissue, Brown metabolism, Adipose Tissue, Brown pathology, Adipose Tissue, White metabolism, Adipose Tissue, White pathology, Cell Death, Inflammation metabolism, Inflammation pathology
- Abstract
In mammals, white adipose tissue (WAT) stores and releases lipids, whereas brown adipose tissue (BAT) oxidizes lipids to fuel thermogenesis. In obese individuals, WAT undergoes profound changes; it expands, becomes dysfunctional, and develops a low-grade inflammatory state. Importantly, BAT content and activity decline in obese subjects, mainly as a result of the conversion of brown adipocytes to white-like unilocular cells. Here, we show that BAT "whitening" is induced by multiple factors, including high ambient temperature, leptin receptor deficiency, β-adrenergic signaling impairment, and lipase deficiency, each of which is capable of inducing macrophage infiltration, brown adipocyte death, and crown-like structure (CLS) formation. Brown-to-white conversion and increased CLS formation were most marked in BAT from adipose triglyceride lipase ( Atgl )-deficient mice, where, according to transmission electron microscopy, whitened brown adipocytes contained enlarged endoplasmic reticulum, cholesterol crystals, and some degenerating mitochondria, and were surrounded by an increased number of collagen fibrils. Gene expression analysis showed that BAT whitening in Atgl -deficient mice was associated to a strong inflammatory response and NLRP3 inflammasome activation. Altogether, the present findings suggest that converted enlarged brown adipocytes are highly prone to death, which, by promoting inflammation in whitened BAT, may contribute to the typical inflammatory state seen in obesity., (Copyright © 2018 by the American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2018
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13. Hypoxia-inducible lipid droplet-associated protein inhibits adipose triglyceride lipase.
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Padmanabha Das KM, Wechselberger L, Liziczai M, De la Rosa Rodriguez M, Grabner GF, Heier C, Viertlmayr R, Radler C, Lichtenegger J, Zimmermann R, Borst JW, Zechner R, Kersten S, and Oberer M
- Subjects
- Humans, Lipase metabolism, Adipocytes enzymology, Adipose Tissue enzymology, Lipase antagonists & inhibitors, Neoplasm Proteins metabolism, Triglycerides metabolism
- Abstract
Elaborate control mechanisms of intracellular triacylglycerol (TAG) breakdown are critically involved in the maintenance of energy homeostasis. Hypoxia-inducible lipid droplet-associated protein (HILPDA)/hypoxia-inducible gene-2 (Hig-2) has been shown to affect intracellular TAG levels, yet, the underlying molecular mechanisms are unclear. Here, we show that HILPDA inhibits adipose triglyceride lipase (ATGL), the enzyme catalyzing the first step of intracellular TAG hydrolysis. HILPDA shares structural similarity with G0/G1 switch gene 2 (G0S2), an established inhibitor of ATGL. HILPDA inhibits ATGL activity in a dose-dependent manner with an IC
50 value of ∼2 μM. ATGL inhibition depends on the direct physical interaction of both proteins and involves the N-terminal hydrophobic region of HILPDA and the N-terminal patatin domain-containing segment of ATGL. Finally, confocal microscopy combined with Förster resonance energy transfer-fluorescence lifetime imaging microscopy analysis indicated that HILPDA and ATGL colocalize and physically interact intracellularly. These findings provide a rational biochemical explanation for the tissue-specific increased TAG accumulation in HILPDA-overexpressing transgenic mouse models., (Copyright © 2018 by the American Society for Biochemistry and Molecular Biology, Inc.)- Published
- 2018
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14. Skin Barrier Development Depends on CGI-58 Protein Expression during Late-Stage Keratinocyte Differentiation.
- Author
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Grond S, Radner FPW, Eichmann TO, Kolb D, Grabner GF, Wolinski H, Gruber R, Hofer P, Heier C, Schauer S, Rülicke T, Hoefler G, Schmuth M, Elias PM, Lass A, Zechner R, and Haemmerle G
- Subjects
- Animals, Cell Differentiation, Ceramides biosynthesis, Lipase physiology, Mice, Skin embryology, Triglycerides metabolism, 1-Acylglycerol-3-Phosphate O-Acyltransferase physiology, Keratinocytes cytology, Skin metabolism
- Abstract
Adipose triglyceride lipase (ATGL) and its coactivator comparative gene identification-58 (CGI-58) are limiting in cellular triglyceride catabolism. Although ATGL deficiency is compatible with normal skin development, mice globally lacking CGI-58 die postnatally and exhibit a severe epidermal permeability barrier defect, which may originate from epidermal and/or peripheral changes in lipid and energy metabolism. Here, we show that epidermis-specific disruption of CGI-58 is sufficient to provoke a defect in the formation of a functional corneocyte lipid envelope linked to impaired ω-O-acylceramide synthesis. As a result, epidermis-specific CGI-58-deficient mice show severe skin dysfunction, arguing for a tissue autonomous cause of disease development. Defective skin permeability barrier formation in global CGI-58-deficient mice could be reversed via transgenic restoration of CGI-58 expression in differentiated but not basal keratinocytes suggesting that CGI-58 is essential for lipid metabolism in suprabasal epidermal layers. The compatibility of ATGL deficiency with normal epidermal function indicated that CGI-58 may stimulate an epidermal triglyceride lipase beyond ATGL required for the adequate provision of fatty acids as a substrate for ω-O-acylceramide synthesis. Pharmacological inhibition of ATGL enzyme activity similarly reduced triglyceride-hydrolytic activities in wild-type and CGI-58 overexpressing epidermis implicating that CGI-58 participates in ω-O-acylceramide biogenesis independent of its role as a coactivator of epidermal triglyceride catabolism., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
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15. PNPLA1 Deficiency in Mice and Humans Leads to a Defect in the Synthesis of Omega-O-Acylceramides.
- Author
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Grond S, Eichmann TO, Dubrac S, Kolb D, Schmuth M, Fischer J, Crumrine D, Elias PM, Haemmerle G, Zechner R, Lass A, and Radner FPW
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- Animals, Ichthyosis etiology, Lipase deficiency, Mice, Mice, Inbred C57BL, Permeability, Ceramides biosynthesis, Lipase physiology, Skin metabolism
- Abstract
Mutations in PNPLA1 have been identified as causative for autosomal recessive congenital ichthyosis in humans and dogs. So far, the underlying molecular mechanisms are unknown. In this study, we generated and characterized PNPLA1-deficient mice and found that PNPLA1 is crucial for epidermal sphingolipid synthesis. The absence of functional PNPLA1 in mice impaired the formation of omega-O-acylceramides and led to an accumulation of nonesterified omega-hydroxy-ceramides. As a consequence, PNPLA1-deficient mice lacked a functional corneocyte-bound lipid envelope leading to a severe skin barrier defect and premature death of newborn animals. Functional analyses of differentiated keratinocytes from a patient with mutated PNPLA1 demonstrated an identical defect in omega-O-acylceramide synthesis in human cells, indicating that PNPLA1 function is conserved among mammals and indispensable for normal skin physiology. Notably, topical application of epidermal lipids from wild-type onto Pnpla1-mutant mice promoted rebuilding of the corneocyte-bound lipid envelope, indicating that supplementation of ichthyotic skin with omega-O-acylceramides might be a therapeutic approach for the treatment of skin symptoms in individuals affected by omega-O-acylceramide deficiency., Competing Interests: The authors state no conflict of interest., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
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16. Mice lacking lipid droplet-associated hydrolase, a gene linked to human prostate cancer, have normal cholesterol ester metabolism.
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Kory N, Grond S, Kamat SS, Li Z, Krahmer N, Chitraju C, Zhou P, Fröhlich F, Semova I, Ejsing C, Zechner R, Cravatt BF, Farese RV Jr, and Walther TC
- Subjects
- Animals, Cholesterol Esters metabolism, Energy Metabolism genetics, Glucose metabolism, Humans, Macrophages metabolism, Male, Mice, Prostatic Neoplasms metabolism, Prostatic Neoplasms pathology, Serine Proteases metabolism, Triglycerides metabolism, Cholesterol Esters genetics, Lipid Droplets metabolism, Lipid Metabolism genetics, Serine Proteases genetics
- Abstract
Variations in the gene LDAH (C2ORF43), which encodes lipid droplet-associated hydrolase (LDAH), are among few loci associated with human prostate cancer. Homologs of LDAH have been identified as proteins of lipid droplets (LDs). LDs are cellular organelles that store neutral lipids, such as triacylglycerols and sterol esters, as precursors for membrane components and as reservoirs of metabolic energy. LDAH is reported to hydrolyze cholesterol esters and to be important in macrophage cholesterol ester metabolism. Here, we confirm that LDAH is localized to LDs in several model systems. We generated a murine model in which Ldah is disrupted but found no evidence for a major function of LDAH in cholesterol ester or triacylglycerol metabolism in vivo, nor a role in energy or glucose metabolism. Our data suggest that LDAH is not a major cholesterol ester hydrolase, and an alternative metabolic function may be responsible for its possible effect on development of prostate cancer., (Copyright © 2017 by the American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2017
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17. ATGL and CGI-58 are lipid droplet proteins of the hepatic stellate cell line HSC-T6.
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Eichmann TO, Grumet L, Taschler U, Hartler J, Heier C, Woblistin A, Pajed L, Kollroser M, Rechberger G, Thallinger GG, Zechner R, Haemmerle G, Zimmermann R, and Lass A
- Subjects
- Adipocytes metabolism, Animals, Cell Line, Lipolysis, Lipoproteins metabolism, Membrane Proteins metabolism, Perilipin-2, Proteome metabolism, Rats, Retinol-Binding Proteins metabolism, Retinyl Esters, Triglycerides metabolism, Vitamin A pharmacology, Acyltransferases metabolism, Hepatic Stellate Cells metabolism, Lipase metabolism, Lipid Droplets metabolism
- Abstract
Lipid droplets (LDs) of hepatic stellate cells (HSCs) contain large amounts of vitamin A [in the form of retinyl esters (REs)] as well as other neutral lipids such as TGs. During times of insufficient vitamin A availability, RE stores are mobilized to ensure a constant supply to the body. To date, little is known about the enzymes responsible for the hydrolysis of neutral lipid esters, in particular of REs, in HSCs. In this study, we aimed to identify LD-associated neutral lipid hydrolases by a proteomic approach using the rat stellate cell line HSC-T6. First, we loaded cells with retinol and FAs to promote lipid synthesis and deposition within LDs. Then, LDs were isolated and lipid composition and the LD proteome were analyzed. Among other proteins, we found perilipin 2, adipose TG lipase (ATGL), and comparative gene identification-58 (CGI-58), known and established LD proteins. Bioinformatic search of the LD proteome for α/β-hydrolase fold-containing proteins revealed no yet uncharacterized neutral lipid hydrolases. In in vitro activity assays, we show that rat (r)ATGL, coactivated by rat (r)CGI-58, efficiently hydrolyzes TGs and REs. These findings suggest that rATGL and rCGI-58 are LD-resident proteins in HSCs and participate in the mobilization of both REs and TGs., (Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2015
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18. Fasting-induced G0/G1 switch gene 2 and FGF21 expression in the liver are under regulation of adipose tissue derived fatty acids.
- Author
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Jaeger D, Schoiswohl G, Hofer P, Schreiber R, Schweiger M, Eichmann TO, Pollak NM, Poecher N, Grabner GF, Zierler KA, Eder S, Kolb D, Radner FP, Preiss-Landl K, Lass A, Zechner R, Kershaw EE, and Haemmerle G
- Subjects
- Animals, Blotting, Western, Diet, High-Fat adverse effects, Disease Models, Animal, Enzyme-Linked Immunosorbent Assay, Fatty Liver metabolism, Fatty Liver pathology, Fibroblast Growth Factors biosynthesis, Genes, Switch, Liver ultrastructure, Mice, Mice, Transgenic, Microscopy, Electron, RNA genetics, Real-Time Polymerase Chain Reaction, Adipose Tissue metabolism, Fasting metabolism, Fatty Acids metabolism, Fatty Liver genetics, Fibroblast Growth Factors genetics, Gene Expression Regulation, Liver metabolism
- Abstract
Background & Aims: Adipose tissue (AT)-derived fatty acids (FAs) are utilized for hepatic triacylglycerol (TG) generation upon fasting. However, their potential impact as signaling molecules is not established. Herein we examined the role of exogenous AT-derived FAs in the regulation of hepatic gene expression by investigating mice with a defect in AT-derived FA supply to the liver., Methods: Plasma FA levels, tissue TG hydrolytic activities and lipid content were determined in mice lacking the lipase co-activator comparative gene identification-58 (CGI-58) selectively in AT (CGI-58-ATko) applying standard protocols. Hepatic expression of lipases, FA oxidative genes, transcription factors, ER stress markers, hormones and cytokines were determined by qRT-PCR, Western blotting and ELISA., Results: Impaired AT-derived FA supply upon fasting of CGI-58-ATko mice causes a marked defect in liver PPARα-signaling and nuclear CREBH translocation. This severely reduced the expression of respective target genes such as the ATGL inhibitor G0/G1 switch gene-2 (G0S2) and the endocrine metabolic regulator FGF21. These changes could be reversed by lipid administration and raising plasma FA levels. Impaired AT-lipolysis failed to induce hepatic G0S2 expression in fasted CGI-58-ATko mice leading to enhanced ATGL-mediated TG-breakdown strongly reducing hepatic TG deposition. On high fat diet, impaired AT-lipolysis counteracts hepatic TG accumulation and liver stress linked to improved systemic insulin sensitivity., Conclusions: AT-derived FAs are a critical regulator of hepatic fasting gene expression required for the induction of G0S2-expression in the liver to control hepatic TG-breakdown. Interfering with AT-lipolysis or hepatic G0S2 expression represents an effective strategy for the treatment of hepatic steatosis., (Copyright © 2015 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)
- Published
- 2015
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19. The tissue distribution of lipoprotein lipase determines where chylomicrons bind.
- Author
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Savonen R, Hiden M, Hultin M, Zechner R, Levak-Frank S, Olivecrona G, and Olivecrona T
- Subjects
- Animals, Chylomicrons genetics, Humans, Lipoprotein Lipase genetics, Mice, Mice, Transgenic, Organ Specificity genetics, Rats, Chylomicrons metabolism, Lipoprotein Lipase metabolism, Muscle, Skeletal metabolism, Myocardium metabolism
- Abstract
To determine the role of LPL for binding of lipoproteins to the vascular endothelium, and for the distribution of lipids from lipoproteins, four lines of induced mutant mice were used. Rat chylomicrons labeled in vivo with [(14)C]oleic acid (primarily in TGs, providing a tracer for lipolysis) and [(3)H]retinol (primarily in ester form, providing a tracer for the core lipids) were injected. TG label was cleared more rapidly than core label. There were no differences between the mouse lines in the rate at which core label was cleared. Two minutes after injection, about 5% of the core label, and hence chylomicron particles, were in the heart of WT mice. In mice that expressed LPL only in skeletal muscle, and had much reduced levels of LPL in the heart, binding of chylomicrons was reduced to 1%, whereas in mice that expressed LPL only in the heart, the binding was increased to over 10%. The same patterns of distribution were evident at 20 min when most of the label had been cleared. Thus, the amount of LPL expressed in muscle and heart governed both the binding of chylomicron particles and the assimilation of chylomicron lipids in the tissue., (Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2015
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20. Fibroblast growth factor 21 is induced upon cardiac stress and alters cardiac lipid homeostasis.
- Author
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Brahma MK, Adam RC, Pollak NM, Jaeger D, Zierler KA, Pöcher N, Schreiber R, Romauch M, Moustafa T, Eder S, Ruelicke T, Preiss-Landl K, Lass A, Zechner R, and Haemmerle G
- Subjects
- Animals, Biological Transport, Cell Line, Energy Metabolism, Fasting metabolism, Fatty Acids metabolism, Female, Glucose metabolism, Lipase deficiency, Male, Mice, Mice, Transgenic, Muscle Fibers, Skeletal metabolism, Organ Specificity, Oxidation-Reduction, RNA, Messenger genetics, RNA, Messenger metabolism, Rats, Endoplasmic Reticulum Stress, Fibroblast Growth Factors genetics, Homeostasis, Myocardium cytology, Myocardium metabolism, Transcriptional Activation, Triglycerides metabolism
- Abstract
Fibroblast growth factor 21 (FGF21) is a PPARα-regulated gene elucidated in the liver of PPARα-deficient mice or PPARα agonist-treated mice. Mice globally lacking adipose triglyceride lipase (ATGL) exhibit a marked defect in TG catabolism associated with impaired PPARα-activated gene expression in the heart and liver, including a drastic reduction in hepatic FGF21 mRNA expression. Here we show that FGF21 mRNA expression is markedly increased in the heart of ATGL-deficient mice accompanied by elevated expression of endoplasmic reticulum (ER) stress markers, which can be reversed by reconstitution of ATGL expression in cardiac muscle. In line with this assumption, the induction of ER stress increases FGF21 mRNA expression in H9C2 cardiomyotubes. Cardiac FGF21 expression was also induced upon fasting of healthy mice, implicating a role of FGF21 in cardiac energy metabolism. To address this question, we generated and characterized mice with cardiac-specific overexpression of FGF21 (CM-Fgf21). FGF21 was efficiently secreted from cardiomyocytes of CM-Fgf21 mice, which moderately affected cardiac TG homeostasis, indicating a role for FGF21 in cardiac energy metabolism. Together, our results show that FGF21 expression is activated upon cardiac ER stress linked to defective lipolysis and that a persistent increase in circulating FGF21 levels interferes with cardiac and whole body energy homeostasis., (Copyright © 2014 by the American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2014
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21. The impact of genetic stress by ATGL deficiency on the lipidome of lipid droplets from murine hepatocytes.
- Author
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Chitraju C, Trötzmüller M, Hartler J, Wolinski H, Thallinger GG, Haemmerle G, Zechner R, Zimmermann R, Köfeler HC, and Spener F
- Subjects
- Animals, Gene Deletion, Hepatocytes chemistry, Lipase genetics, Lipid Metabolism, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Particle Size, DNA Damage, Hepatocytes metabolism, Lipase deficiency, Lipids
- Abstract
We showed earlier that nutritional stress like starvation or high-fat diet resulted in phenotypic changes in the lipidomes of hepatocyte lipid droplets (LDs), representative for the pathophysiological status of the mouse model. Here we extend our former study by adding genetic stress due to knockout (KO) of adipocyte triglyceride lipase (ATGL), the rate limiting enzyme in LD lipolysis. An intervention trial for 6 weeks with male wild-type (WT) and ATGL-KO mice was carried out; both genotypes were fed lab chow or were exposed to short-time starvation. Isolated LDs were analyzed by LC-MS/MS. Triacylglycerol, diacylglycerol, and phosphatidylcholine lipidomes, in that order, provided the best phenotypic signatures characteristic for respective stresses applied to the animals. This was evidenced at lipid species level by principal component analysis, calculation of average values for chain-lengths and numbers of double bonds, and by visualization in heat maps. Structural backgrounds for analyses and metabolic relationships were elaborated at lipid molecular species level. Relating our lipidomic data to nonalcoholic fatty liver diseases of nutritional and genetic etiologies with or without accompanying insulin resistance, phenotypic distinction in hepatocyte LDs dependent on insulin status emerged. Taken together, lipidomes of hepatocyte LDs are sensitive responders to nutritional and genetic stress.
- Published
- 2013
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22. Cardiac-specific overexpression of perilipin 5 provokes severe cardiac steatosis via the formation of a lipolytic barrier.
- Author
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Pollak NM, Schweiger M, Jaeger D, Kolb D, Kumari M, Schreiber R, Kolleritsch S, Markolin P, Grabner GF, Heier C, Zierler KA, Rülicke T, Zimmermann R, Lass A, Zechner R, and Haemmerle G
- Subjects
- 1-Acylglycerol-3-Phosphate O-Acyltransferase genetics, 1-Acylglycerol-3-Phosphate O-Acyltransferase metabolism, Animals, COS Cells, Cardiomyopathies genetics, Energy Metabolism genetics, Energy Metabolism physiology, Immunoblotting, Intracellular Signaling Peptides and Proteins genetics, Lipase genetics, Lipase metabolism, Lipolysis genetics, Mice, Mice, Transgenic, Muscle Proteins genetics, Myocardium metabolism, Myocardium pathology, Triglycerides metabolism, Cardiomyopathies metabolism, Intracellular Signaling Peptides and Proteins metabolism, Lipolysis physiology, Muscle Proteins metabolism
- Abstract
Cardiac triacylglycerol (TG) catabolism critically depends on the TG hydrolytic activity of adipose triglyceride lipase (ATGL). Perilipin 5 (Plin5) is expressed in cardiac muscle (CM) and has been shown to interact with ATGL and its coactivator comparative gene identification-58 (CGI-58). Furthermore, ectopic Plin5 expression increases cellular TG content and Plin5-deficient mice exhibit reduced cardiac TG levels. In this study we show that mice with cardiac muscle-specific overexpression of perilipin 5 (CM-Plin5) massively accumulate TG in CM, which is accompanied by moderately reduced fatty acid (FA) oxidizing gene expression levels. Cardiac lipid droplet (LD) preparations from CM of CM-Plin5 mice showed reduced ATGL- and hormone-sensitive lipase-mediated TG mobilization implying that Plin5 overexpression restricts cardiac lipolysis via the formation of a lipolytic barrier. To test this hypothesis, we analyzed TG hydrolytic activities in preparations of Plin5-, ATGL-, and CGI-58-transfected cells. In vitro ATGL-mediated TG hydrolysis of an artificial micellar TG substrate was not inhibited by the presence of Plin5, whereas Plin5-coated LDs were resistant toward ATGL-mediated TG catabolism. These findings strongly suggest that Plin5 functions as a lipolytic barrier to protect the cardiac TG pool from uncontrolled TG mobilization and the excessive release of free FAs.
- Published
- 2013
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23. G0/G1 switch gene-2 regulates human adipocyte lipolysis by affecting activity and localization of adipose triglyceride lipase.
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Schweiger M, Paar M, Eder C, Brandis J, Moser E, Gorkiewicz G, Grond S, Radner FP, Cerk I, Cornaciu I, Oberer M, Kersten S, Zechner R, Zimmermann R, and Lass A
- Subjects
- 3T3-L1 Cells, Animals, Cell Cycle Proteins genetics, Cell Differentiation, Cells, Cultured, Cysteine Endopeptidases genetics, Humans, Immunoblotting, In Vitro Techniques, Lipase genetics, Lipolysis genetics, Mice, Mice, Inbred C57BL, Microscopy, Fluorescence, Mutagenesis, Site-Directed, Adipocytes metabolism, Cell Cycle Proteins metabolism, Cysteine Endopeptidases metabolism, Lipase metabolism, Lipolysis physiology
- Abstract
The hydrolysis of triglycerides in adipocytes, termed lipolysis, provides free fatty acids as energy fuel. Murine lipolysis largely depends on the activity of adipose triglyceride lipase (ATGL), which is regulated by two proteins annotated as comparative gene identification-58 (CGI-58) and G0/G1 switch gene-2 (G0S2). CGI-58 activates and G0S2 inhibits ATGL activity. In contrast to mice, the functional role of G0S2 in human adipocyte lipolysis is poorly characterized. Here we show that overexpression or silencing of G0S2 in human SGBS adipocytes decreases and increases lipolysis, respectively. Human G0S2 is upregulated during adipocyte differentiation and inhibits ATGL activity in a dose-dependent manner. Interestingly, C-terminally truncated ATGL mutants, which fail to localize to lipid droplets, translocate to the lipid droplet upon coexpression with G0S2, suggesting that G0S2 anchors ATGL to lipid droplets independent of ATGL's C-terminal lipid binding domain. Taken together, our results indicate that G0S2 also regulates human lipolysis by affecting enzyme activity and intracellular localization of ATGL. Increased lipolysis is known to contribute to the pathogenesis of insulin resistance, and G0S2 expression has been shown to be reduced in poorly controlled type 2 diabetic patients. Our data indicate that downregulation of G0S2 in adipose tissue could represent one of the underlying causes leading to increased lipolysis in the insulin-resistant state.
- Published
- 2012
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24. Lipidomic analysis of lipid droplets from murine hepatocytes reveals distinct signatures for nutritional stress.
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Chitraju C, Trötzmüller M, Hartler J, Wolinski H, Thallinger GG, Lass A, Zechner R, Zimmermann R, Köfeler HC, and Spener F
- Subjects
- Animals, Diet, High-Fat, Diglycerides metabolism, Fasting, Hepatocytes chemistry, Lipase metabolism, Liver chemistry, Mice, Phosphatidylcholines metabolism, Phosphatidylethanolamine N-Methyltransferase metabolism, Triglycerides metabolism, Hepatocytes metabolism, Lipids analysis, Liver metabolism, Stress, Physiological
- Abstract
Liver steatosis can be induced by fasting or high-fat diet. We investigated by lipidomic analysis whether such metabolic states are reflected in the lipidome of hepatocyte lipid droplets (LDs) from mice fed normal chow diet (FED), fasted (FAS), or fed a high-fat diet (HFD). LC-MS/MS at levels of lipid species profiles and of lipid molecular species uncovered a FAS phenotype of LD enriched in triacylglycerol (TG) molecular species with very long-chain (VLC)-PUFA residues and an HFD phenotype with less unsaturated TG species in addition to characteristic lipid marker species. Nutritional stress did not result in dramatic structural alterations in diacylglycerol (DG) and phospholipid (PL) classes. Moreover, molecular species of bulk TG and of DG indicated concomitant de novo TG synthesis and lipase-catalyzed degradation to be active in LDs. DG species with VLC-PUFA residues would be preferred precursors for phosphatidylcholine (PC) species, the others for TG molecular species. In addition, molecular species of PL classes fitted the hepatocyte Kennedy and phosphatidylethanolamine methyltransferase pathways. We demonstrate that lipidomic analysis of LDs enables phenotyping of nutritional stress. TG species are best suited for such phenotyping, whereas structural analysis of TG, DG, and PL molecular species provides metabolic insights.
- Published
- 2012
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25. DGAT enzymes are required for triacylglycerol synthesis and lipid droplets in adipocytes.
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Harris CA, Haas JT, Streeper RS, Stone SJ, Kumari M, Yang K, Han X, Brownell N, Gross RW, Zechner R, and Farese RV Jr
- Subjects
- Animals, Blotting, Western, Diacylglycerol O-Acyltransferase genetics, Enzyme-Linked Immunosorbent Assay, Fluorescent Antibody Technique, Macrophages metabolism, Mass Spectrometry, Mice, Mice, Knockout, Microscopy, Electron, Transmission, Triglycerides genetics, Adipocytes enzymology, Adipocytes metabolism, Diacylglycerol O-Acyltransferase metabolism, Triglycerides biosynthesis
- Abstract
The total contribution of the acyl CoA:diacylglycerol acyltransferase (DGAT) enzymes, DGAT1 and DGAT2, to mammalian triacylglycerol (TG) synthesis has not been determined. Similarly, whether DGAT enzymes are required for lipid droplet (LD) formation is unknown. In this study, we examined the requirement for DGAT enzymes in TG synthesis and LDs in differentiated adipocytes with genetic deletions of DGAT1 and DGAT2. Adipocytes with a single deletion of either enzyme were capable of TG synthesis and LD formation. In contrast, adipocytes with deletions of both DGATs were severely lacking in TG and did not have LDs, indicating that DGAT1 and DGAT2 account for nearly all TG synthesis in adipocytes and appear to be required for LD formation during adipogenesis. DGAT enzymes were not absolutely required for LD formation in mammalian cells, however; macrophages deficient in both DGAT enzymes were able to form LDs when incubated with cholesterol-rich lipoproteins. Although adipocytes lacking both DGATs had no TG or LDs, they were fully differentiated by multiple criteria. Our findings show that DGAT1 and DGAT2 account for the vast majority of TG synthesis in mice, and DGAT function is required for LDs in adipocytes, but not in all cell types.
- Published
- 2011
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26. Pnpla3/Adiponutrin deficiency in mice does not contribute to fatty liver disease or metabolic syndrome.
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Basantani MK, Sitnick MT, Cai L, Brenner DS, Gardner NP, Li JZ, Schoiswohl G, Yang K, Kumari M, Gross RW, Zechner R, and Kershaw EE
- Subjects
- Adipose Tissue metabolism, Animals, Energy Metabolism, Fatty Liver etiology, Male, Metabolic Syndrome etiology, Mice, Mice, Knockout, Triglycerides metabolism, Fatty Liver metabolism, Metabolic Syndrome metabolism, Phospholipases A2, Calcium-Independent deficiency
- Abstract
PNPLA3 (adiponutrin, calcium-independent phospholipase A(2) epsilon [iPLA(2)ε]) is an adipose-enriched, nutritionally regulated protein that belongs to the patatin-like phospholipase domain containing (PNPLA) family of lipid metabolizing proteins. Genetic variations in the human PNPLA3 gene (i.e., the rs738409 I148M allele) has been strongly and repeatedly associated with fatty liver disease. Although human PNPLA3 has triacylglycerol (TAG) hydrolase and transacylase activities in vitro, its in vivo function and physiological relevance remain controversial. The objective of this study was to determine the metabolic consequences of global targeted deletion of the Pnpla3 gene in mice. We found that Pnpla3 mRNA expression is altered in adipose tissue and liver in response to acute and chronic nutritional challenges. However, global targeted deletion of the Pnpla3 gene in mice did not affect TAG hydrolysis, nor did it influence energy/glucose/lipid homoeostasis or hepatic steatosis/injury. Experimental interventions designed to increase Pnpla3 expression (refeeding, high-sucrose diet, diet-induced obesity, and liver X receptor agonism) likewise failed to reveal differences in the above-mentioned metabolic phenotypes. Expression of the Pnpla3 paralog, Pnpla5, was increased in adipose tissue but not in liver of Pnpla3-deficient mice, but compensatory regulation of genes involved in TAG metabolism was not identified. Together these data argue against a role for Pnpla3 loss-of-function in fatty liver disease or metabolic syndrome in mice.
- Published
- 2011
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27. Cholesteryl ester hydrolase activity is abolished in HSL-/- macrophages but unchanged in macrophages lacking KIAA1363.
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Buchebner M, Pfeifer T, Rathke N, Chandak PG, Lass A, Schreiber R, Kratzer A, Zimmermann R, Sattler W, Koefeler H, Fröhlich E, Kostner GM, Birner-Gruenberger R, Chiang KP, Haemmerle G, Zechner R, Levak-Frank S, Cravatt B, and Kratky D
- Subjects
- Animals, Atherosclerosis genetics, Atherosclerosis metabolism, COS Cells, Chlorocebus aethiops, Diglycerides metabolism, Hydrolysis, Macrophages metabolism, Mice, Mice, Inbred Strains, Microscopy, Fluorescence, Serine Proteases metabolism, Sterol Esterase metabolism, Transfection, Cholesterol Esters metabolism, Macrophages enzymology, Serine Proteases genetics, Sterol Esterase genetics
- Abstract
Cholesteryl ester (CE) accumulation in macrophages represents a crucial event during foam cell formation, a hallmark of atherogenesis. Here we investigated the role of two previously described CE hydrolases, hormone-sensitive lipase (HSL) and KIAA1363, in macrophage CE hydrolysis. HSL and KIAA1363 exhibited marked differences in their abilities to hydrolyze CE, triacylglycerol (TG), diacylglycerol (DG), and 2-acetyl monoalkylglycerol ether (AcMAGE), a precursor for biosynthesis of platelet-activating factor (PAF). HSL efficiently cleaved all four substrates, whereas KIAA1363 hydrolyzed only AcMAGE. This contradicts previous studies suggesting that KIAA1363 is a neutral CE hydrolase. Macrophages of KIAA1363(-/-) and wild-type mice exhibited identical neutral CE hydrolase activity, which was almost abolished in tissues and macrophages of HSL(-/-) mice. Conversely, AcMAGE hydrolase activity was diminished in macrophages and some tissues of KIAA1363(-/-) but unchanged in HSL(-/-) mice. CE turnover was unaffected in macrophages lacking KIAA1363 and HSL, whereas cAMP-dependent cholesterol efflux was influenced by HSL but not by KIAA1363. Despite decreased CE hydrolase activities, HSL(-/-) macrophages exhibited CE accumulation similar to wild-type (WT) macrophages. We conclude that additional enzymes must exist that cooperate with HSL to regulate CE levels in macrophages. KIAA1363 affects AcMAGE hydrolase activity but is of minor importance as a direct CE hydrolase in macrophages.
- Published
- 2010
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28. Adipose triglyceride lipase plays a key role in the supply of the working muscle with fatty acids.
- Author
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Schoiswohl G, Schweiger M, Schreiber R, Gorkiewicz G, Preiss-Landl K, Taschler U, Zierler KA, Radner FP, Eichmann TO, Kienesberger PC, Eder S, Lass A, Haemmerle G, Alsted TJ, Kiens B, Hoefler G, Zechner R, and Zimmermann R
- Subjects
- Animals, Carbohydrates blood, Carboxylic Ester Hydrolases deficiency, Carboxylic Ester Hydrolases genetics, Energy Metabolism, Female, Gene Knockout Techniques, Glycogen metabolism, Lipase, Lipids blood, Liver metabolism, Locomotion, Male, Mice, Muscles cytology, Muscles physiology, Mutation, Physical Conditioning, Animal, Rest, Carboxylic Ester Hydrolases metabolism, Fatty Acids metabolism, Muscles metabolism
- Abstract
FAs are mobilized from triglyceride (TG) stores during exercise to supply the working muscle with energy. Mice deficient for adipose triglyceride lipase (ATGL-ko) exhibit defective lipolysis and accumulate TG in adipose tissue and muscle, suggesting that ATGL deficiency affects energy availability and substrate utilization in working muscle. In this study, we investigated the effect of moderate treadmill exercise on blood energy metabolites and liver glycogen stores in mice lacking ATGL. Because ATGL-ko mice exhibit massive accumulation of TG in the heart and cardiomyopathy, we also investigated a mouse model lacking ATGL in all tissues except cardiac muscle (ATGL-ko/CM). In contrast to ATGL-ko mice, these mice did not accumulate TG in the heart and had normal life expectancy. Exercise experiments revealed that ATGL-ko and ATGL-ko/CM mice are unable to increase circulating FA levels during exercise. The reduced availability of FA for energy conversion led to rapid depletion of liver glycogen stores and hypoglycemia. Together, our studies suggest that ATGL-ko mice cannot adjust circulating FA levels to the increased energy requirements of the working muscle, resulting in an increased use of carbohydrates for energy conversion. Thus, ATGL activity is required for proper energy supply of the skeletal muscle during exercise.
- Published
- 2010
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29. Esterase 22 and beta-glucuronidase hydrolyze retinoids in mouse liver.
- Author
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Schreiber R, Taschler U, Wolinski H, Seper A, Tamegger SN, Graf M, Kohlwein SD, Haemmerle G, Zimmermann R, Zechner R, and Lass A
- Subjects
- Animals, COS Cells, Chlorocebus aethiops, Hydrolysis, Liver enzymology, Male, Mice, Mice, Inbred C57BL, Carboxylic Ester Hydrolases metabolism, Glucuronidase metabolism, Liver metabolism, Retinoids metabolism
- Abstract
Excess dietary vitamin A is esterified with fatty acids and stored in the form of retinyl ester (RE) predominantly in the liver. According to the requirements of the body, liver RE stores are hydrolyzed and retinol is delivered to peripheral tissues. The controlled mobilization of retinol ensures a constant supply of the body with the vitamin. Currently, the enzymes catalyzing liver RE hydrolysis are unknown. In this study, we identified mouse esterase 22 (Es22) as potent RE hydrolase highly expressed in the liver, particularly in hepatocytes. The enzyme is located exclusively at the endoplasmic reticulum (ER), implying that it is not involved in the mobilization of RE present in cytosolic lipid droplets. Nevertheless, cell culture experiments revealed that overexpression of Es22 attenuated the formation of cellular RE stores, presumably by counteracting retinol esterification at the ER. Es22 was previously shown to form a complex with beta-glucuronidase (Gus). Our studies revealed that Gus colocalizes with Es22 at the ER but does not affect its RE hydrolase activity. Interestingly, however, Gus was capable of hydrolyzing the naturally occurring vitamin A metabolite retinoyl beta-glucuronide. In conclusion, our observations implicate that both Es22 and Gus play a role in liver retinoid metabolism.
- Published
- 2009
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30. Adipose tissue as a source of nicotinamide N-methyltransferase and homocysteine.
- Author
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Riederer M, Erwa W, Zimmermann R, Frank S, and Zechner R
- Subjects
- 3T3-L1 Cells, Adipocytes drug effects, Adipogenesis, Adipose Tissue drug effects, Animals, Enzyme Inhibitors pharmacology, Female, Gene Expression Regulation, Enzymologic, Humans, Hypolipidemic Agents pharmacology, Male, Mice, Mice, Inbred C57BL, Niacin pharmacology, Niacinamide analogs & derivatives, Niacinamide pharmacology, Nicotinamide N-Methyltransferase antagonists & inhibitors, Nicotinamide N-Methyltransferase genetics, RNA, Messenger metabolism, Time Factors, Adipocytes enzymology, Adipose Tissue enzymology, Homocysteine metabolism, Nicotinamide N-Methyltransferase metabolism
- Abstract
Nicotinamide N-methyltransferase (NNMT) catalyses the conversion of nicotinamide to 1-methylnicotinamide and plays an important role in hepatic detoxification reactions. Here we show that, in addition to the liver, 3T3-L1 adipocytes as well as human and murine adipose tissue explants express high amounts of enzymatically active NNMT. NNMT mRNA levels and enzyme activity increased in 3T3-L1 cells in a differentiation-dependent manner. Homocysteine, the atherogenic product of the NNMT-catalyzed reaction, was secreted from 3T3-L1 cells or adipose tissue cultures. Homocysteine release increased during 3T3-L1 differentiation and was reduced when adipose tissue was treated with the NNMT inhibitor 1-methylnicotinamide. Nicotinic acid (NA), a widely used drug to lower elevated plasma lipid levels, induced NNMT enzyme activity in white adipose tissue of mice. In tissue culture nicotinamide treatment led to an increase in adipose tissue homocysteine secretion. These data support the concept that adipose tissue NNMT contributes to the increased plasma homocysteine levels in patients treated with NA.
- Published
- 2009
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31. Mammalian patatin domain containing proteins: a family with diverse lipolytic activities involved in multiple biological functions.
- Author
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Kienesberger PC, Oberer M, Lass A, and Zechner R
- Subjects
- Animals, Humans, Keratinocytes enzymology, Lipase chemistry, Lipase classification, Lipase genetics, Mammals classification, Mammals genetics, Triglycerides metabolism, Biological Phenomena, Lipase metabolism, Lipolysis, Mammals metabolism
- Abstract
The human genome expresses nine patatin-like phospholipase domain containing proteins (PNPLA1-9). Members of this family share a protein domain discovered initially in patatin, the most abundant protein of the potato tuber. Patatin is a lipid hydrolase with an unusual folding topology that differs from classical lipases. Mammalian PNPLAs include lipid hydrolases with specificities for diverse substrates such as triacylglycerols, phospholipids, and retinol esters. Analysis of induced mutant mouse models and the clinical phenotype of patients with mutations revealed important insights into the physiological role of several members of the PNPLA family. This review aims to summarize current knowledge of PNPLA proteins and to document their emerging importance in lipid and energy homeostasis.
- Published
- 2009
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32. Adipose triglyceride lipase and the lipolytic catabolism of cellular fat stores.
- Author
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Zechner R, Kienesberger PC, Haemmerle G, Zimmermann R, and Lass A
- Subjects
- Adipose Tissue metabolism, Animals, Fatty Acids metabolism, Humans, Hydrolases chemistry, Hydrolysis, Lipids chemistry, Lipolysis, Metabolism, Mice, Monoacylglycerol Lipases metabolism, Mutation, Phenotype, Sterol Esterase metabolism, Adipose Tissue enzymology, Lipase metabolism
- Abstract
Fatty acids (FAs) are essential components of all lipid classes and pivotal substrates for energy production in all vertebrates. Additionally, they act directly or indirectly as signaling molecules and, when bonded to amino acid side chains of peptides, anchor proteins in biological membranes. In vertebrates, FAs are predominantly stored in the form of triacylglycerol (TG) within lipid droplets of white adipose tissue. Lipid droplet-associated TGs are also found in most nonadipose tissues, including liver, cardiac muscle, and skeletal muscle. The mobilization of FAs from all fat depots depends on the activity of TG hydrolases. Currently, three enzymes are known to hydrolyze TG, the well-studied hormone-sensitive lipase (HSL) and monoglyceride lipase (MGL), discovered more than 40 years ago, as well as the relatively recently identified adipose triglyceride lipase (ATGL). The phenotype of HSL- and ATGL-deficient mice, as well as the disease pattern of patients with defective ATGL activity (due to mutation in ATGL or in the enzyme's activator, CGI-58), suggest that the consecutive action of ATGL, HSL, and MGL is responsible for the complete hydrolysis of a TG molecule. The complex regulation of these enzymes by numerous, partially uncharacterized effectors creates the "lipolysome," a complex metabolic network that contributes to the control of lipid and energy homeostasis. This review focuses on the structure, function, and regulation of lipolytic enzymes with a special emphasis on ATGL.
- Published
- 2009
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33. LPL-mediated lipolysis of VLDL induces an upregulation of AU-rich mRNAs and an activation of HuR in endothelial cells.
- Author
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Tschernatsch MM, Mlecnik B, Trajanoski Z, Zechner R, and Zimmermann R
- Subjects
- Animals, Antigens, Surface drug effects, Aorta, Thoracic cytology, Aorta, Thoracic metabolism, Blotting, Northern, Blotting, Western, Cells, Cultured, Cholesterol, VLDL drug effects, ELAV Proteins, ELAV-Like Protein 1, Endothelium, Vascular cytology, Heterogeneous Nuclear Ribonucleoprotein D0, Heterogeneous-Nuclear Ribonucleoprotein D drug effects, Humans, RNA, Messenger genetics, RNA-Binding Proteins drug effects, Swine, Antigens, Surface genetics, Cholesterol, VLDL metabolism, Endothelium, Vascular metabolism, Heterogeneous-Nuclear Ribonucleoprotein D genetics, Lipolysis drug effects, Lipoprotein Lipase pharmacology, RNA-Binding Proteins genetics, Up-Regulation drug effects
- Abstract
Lipoprotein lipase (LPL), the major enzyme for hydrolysis of circulating triglyceride-rich lipoproteins, is bound to the luminal surface of capillary endothelial cells. Products of LPL-mediated lipolysis, such as free fatty acids (FFA) and lipoprotein-remnants, can affect endothelial function and gene expression, and hence vascular homeostasis. In this study we tried to identify lipolysis-induced mRNAs in porcine aortic endothelial cells (ECAP) using a cDNA subtraction method. cDNA obtained from ECAP incubated with LPL and VLDL was subtracted from cDNA from cells cultured under control conditions. Analysis of the identified sequences revealed an upregulation of several mRNAs with adenine and uracil-rich elements (ARE) in their 3'-untranslated regions, such as IL-8, ESM-1 and VCAM-1. HuR, a ubiquitously expressed RNA-binding protein, is known to stabilize ARE-harboring mRNAs. Therefore, we investigated whether HuR is involved in this process and found that lipolysis induced an increased polysomal localization of HuR, which is typical for its activation pathway. In addition, the mRNAs for GM-CSF and TNF-alpha - established ARE-containing targets for HuR-mediated regulation - were upregulated by LPL-mediated lipolysis in ECAP. Differential expression of AU-rich mRNAs in response to LPL-mediated lipolysis might have an impact on physiological processes regulating lipid metabolism or pathophysiological processes promoting endothelial dysfunction and atherogenesis.
- Published
- 2006
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34. Endothelial and lipoprotein lipases in human and mouse placenta.
- Author
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Lindegaard ML, Olivecrona G, Christoffersen C, Kratky D, Hannibal J, Petersen BL, Zechner R, Damm P, and Nielsen LB
- Subjects
- Animals, Biopsy, Blotting, Western, Chromatography, Affinity, Dimerization, Heparin chemistry, Humans, Immunohistochemistry, In Situ Hybridization, Mice, Mice, Transgenic, Phospholipids chemistry, RNA, Messenger metabolism, Sepharose chemistry, Sepharose pharmacology, Sodium Chloride pharmacology, Species Specificity, Triglycerides chemistry, Trophoblasts metabolism, Endothelium enzymology, Lipoprotein Lipase chemistry, Placenta enzymology
- Abstract
Placenta expresses various lipase activities. However, a detailed characterization of the involved genes and proteins is lacking. In this study, we compared the expression of endothelial lipase (EL) and LPL in human term placenta. When placental protein extracts were separated by heparin-Sepharose affinity chromatography, the EL protein eluted as a single peak without detectable phospholipid or triglyceride (TG) lipase activity. The major portion of LPL protein eluted slightly after EL. This peak also had no lipase activity and most likely contained monomeric LPL. Fractions eluting at a higher NaCl concentration contained small amounts of LPL protein (most likely dimeric LPL) and had substantial TG lipase activity. In situ hybridization studies showed EL mRNA expression in syncytiotrophoblasts and endothelial cells and LPL mRNA in syncytiotrophoblasts. In contrast, immunohistochemistry showed EL and LPL protein associated with both cell types. In mouse placentas, lack of LPL expression resulted in increased EL mRNA expression. These results suggest that the cellular expression of EL and LPL in human placenta is different. Nevertheless, the two lipases might have overlapping functions in the mouse placenta. Our data also suggest that the major portions of both proteins are stored in an inactive form in human term placenta.
- Published
- 2005
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35. Defective uptake of triglyceride-associated fatty acids in adipose tissue causes the SREBP-1c-mediated induction of lipogenesis.
- Author
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Wagner EM, Kratky D, Haemmerle G, Hrzenjak A, Kostner GM, Steyrer E, and Zechner R
- Subjects
- Animals, Animals, Genetically Modified, CCAAT-Enhancer-Binding Proteins genetics, DNA-Binding Proteins genetics, Diet, Endothelium metabolism, Gene Expression Regulation, Enzymologic physiology, Humans, Insulin metabolism, Leptin metabolism, Lipids biosynthesis, Lipoprotein Lipase genetics, Mice, Muscles metabolism, Sterol Regulatory Element Binding Protein 1, Adipose Tissue metabolism, CCAAT-Enhancer-Binding Proteins metabolism, DNA-Binding Proteins metabolism, Fatty Acids metabolism, Lipoprotein Lipase metabolism, Transcription Factors, Triglycerides metabolism
- Abstract
Lipoprotein lipase (LPL) is the only known enzyme in the capillary endothelium of peripheral tissues that hydrolizes plasma triglycerides and provides fatty acids (FAs) for their subsequent tissue uptake. Previously, we demonstrated that mice that express LPL exclusively in muscle develop essentially normal fat mass despite the absence of LPL and the deprivation of nutritionally derived FAs in adipose tissue (AT). Using this mouse model, we now investigated the metabolic response to LPL deficiency in AT that enables maintenance of normal AT mass. We show that the rate of FA production was 1.8-fold higher in LPL-deficient AT than in control AT. The levels of mRNA and enzymatic activities of important enzymes involved in FA and triglyceride biosynthesis were induced concomitantly. Increased plasma glucose clearing and (14)C-deoxyglucose uptake into LPL-deficient mouse fat pads indicated that glucose provided the carbon source for lipid synthesis. Leptin expression was decreased in LPL-deficient AT. Finally, the induction of de novo FA synthesis in LPL-deficient AT was associated with increased expression and processing of sterol regulatory element binding protein 1 (SREBP-1), together with an increase in INSIG-1 expression. These results suggest that in the absence of LPL in AT, lipogenesis is activated through increased SREBP-1 expression and processing triggered by decreased availability of nutrition-derived FAs, elevated insulin, and low leptin levels.
- Published
- 2004
- Full Text
- View/download PDF
36. Decreased fatty acid esterification compensates for the reduced lipolytic activity in hormone-sensitive lipase-deficient white adipose tissue.
- Author
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Zimmermann R, Haemmerle G, Wagner EM, Strauss JG, Kratky D, and Zechner R
- Subjects
- Acyltransferases genetics, Acyltransferases metabolism, Adipose Tissue chemistry, Animals, Body Weight, CCAAT-Enhancer-Binding Proteins genetics, DNA-Binding Proteins genetics, Esterification, Fatty Acids biosynthesis, Fatty Acids chemistry, Fatty Acids, Nonesterified biosynthesis, Fatty Acids, Nonesterified metabolism, Glucose metabolism, Lipolysis, Male, Mice, Mice, Knockout, RNA, Messenger genetics, RNA, Messenger metabolism, Receptors, Cytoplasmic and Nuclear genetics, Sterol Esterase genetics, Sterol Esterase metabolism, Sterol Regulatory Element Binding Protein 1, Transcription Factors genetics, Adipose Tissue enzymology, Adipose Tissue metabolism, Fatty Acids metabolism, Sterol Esterase deficiency
- Abstract
It has been observed previously that hormone-sensitive lipase-deficient (HSL-ko) mice have reduced white adipose tissue (WAT) stores compared to control mice. These findings contradict the expectation that the decreased lipolytic activity in WAT of HSL-ko mice would cause accumulation of triglycerides (TGs) in that tissue. Here we demonstrate that the cellular TG synthesis in HSL-deficient WAT is markedly reduced due to downregulation of the enzymatic activities of glycerophosphate acyltransferase, dihydroxyacetonphosphate acyltransferase, lysophosphatidate acyltransferase, and diacylglycerol acyltransferase. Fatty acid de novo synthesis is also decreased due to reduced cellular glucose uptake, reduced glucose incorporation into adipose tissue lipids, and reduced activities of acetyl:CoA carboxylase and fatty acid synthase. Finally, the activities of phosphoenolpyruvate carboxykinase (PEPCK), acyl:CoA synthetase (ACS), and glucose 6-phosphate dehydrogenase, the enzymes that provide glycerol-3-phosphate, acyl-CoA, and NADPH for TG synthesis, respectively, are decreased in HSL-ko mice. The reduced expression of the peroxisome proliferator-activated receptor gamma (PPAR gamma) target genes PEPCK, ACS, and aP2, as well as reduced mRNA levels of PPAR gamma itself, suggest the involvement of this transcription factor in the downregulation of lipogenesis. Taken together, these results establish that in the absence of HSL, the reduced NEFA production is counteracted by a drastic reduction of NEFA reesterification that provides sufficient quantities of NEFA for release into the circulation. These metabolic adaptations result in decreased fat mass in HSL-ko mice.
- Published
- 2003
- Full Text
- View/download PDF
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