In Vivo Lipogenesis of Genetically Lean and Fat Chickens: Effects of Nutritional State and Dietary Fat
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Abstract Lipogenesis is a vital but often dysregulated metabolic pathway. We report super-resolution multiplexed vibrational imaging of lipogenesis rates and pathways using isotopically labelled oleic acid and glucose as probes in live adipocytes and hepatocytes. These findings suggest oleic acid inhibits de novo lipogenesis (DNL), but not total lipogenesis, in hepatocytes. No significant effect is seen in adipocytes. These differential effects may be due to alternate regulation of DNL between cell types and could help explain the complicated role oleic acid plays in metabolism.
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Research Article| May 01 1982 Brown-adipose-tissue lipogenesis in starvation: effects of insulin and (_)hydroxycitrate Mary C. Sugden; Mary C. Sugden *Department of Biochemistry, Charing Cross Hospital Medical School, Fulham Palace Road, London, W6 8RF, U.K.†Department of Surgery, Charing Cross Hospital Medical School, Fulham Palace Road, London, W6 8RF, U.K. Search for other works by this author on: This Site PubMed Google Scholar David I. Watts; David I. Watts *Department of Biochemistry, Charing Cross Hospital Medical School, Fulham Palace Road, London, W6 8RF, U.K. Search for other works by this author on: This Site PubMed Google Scholar Christopher E. Marshall; Christopher E. Marshall †Department of Surgery, Charing Cross Hospital Medical School, Fulham Palace Road, London, W6 8RF, U.K. Search for other works by this author on: This Site PubMed Google Scholar James G. McCormack James G. McCormack ††Department of Biochemistry, University of Bristol Medical School, Bristol, BS8 1TD, U.K. Search for other works by this author on: This Site PubMed Google Scholar Biosci Rep (1982) 2 (5): 289–297. https://doi.org/10.1007/BF01115114 Article history Received: April 26 1982 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation Mary C. Sugden, David I. Watts, Christopher E. Marshall, James G. McCormack; Brown-adipose-tissue lipogenesis in starvation: effects of insulin and (_)hydroxycitrate. Biosci Rep 1 May 1982; 2 (5): 289–297. doi: https://doi.org/10.1007/BF01115114 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBioscience Reports Search Advanced Search This content is only available as a PDF. © 1982 The Biochemical Society1982 Article PDF first page preview Close Modal You do not currently have access to this content.
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This study compared the effects of certain metabolites (either singly or in various combinations) and the methods of measuring lipogenesis (using either 14C-acetate or 3H2O incorporation into lipids) on total lipid synthesis and insulin-stimulated total lipid synthesis in the isolated rat hepatocyte. There were quantitative and qualitative differences between 14C-acetate and 3H2O incorporation into lipids; metabolites acutely affected both lipogenesis and insulin-stimulated lipogenesis with either isotope; and insulin's effect on lipogenesis was greater when measured by 14C-acetate incorporation. It is suggested that a particular choice of incubation media and isotope may inadvertently bias a study of insulin-stimulated lipogenesis and that metabolite supply plays a major role in regulating insulin-stimulated lipogenesis.
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Lipid metabolism in liver is complex. In addition to importing and exporting lipid via lipoproteins, hepatocytes can oxidize lipid via fatty acid oxidation, or alternatively, synthesize new lipid via de novo lipogenesis. The net sum of these pathways is dictated by a number of factors, which in certain disease states leads to fatty liver disease. Excess hepatic lipid accumulation is associated with whole body insulin resistance and coronary heart disease. Tools to study lipid metabolism in hepatocytes are useful to understand the role of hepatic lipid metabolism in certain metabolic disorders. In the liver, hepatocytes regulate the breakdown and synthesis of fatty acids via β-fatty oxidation and de novo lipogenesis, respectively. Quantifying metabolism in these pathways provides insight into hepatic lipid handling. Unlike in vitro quantification, using primary hepatocytes, making measurements in vivo is technically challenging and resource intensive. Hence, quantifying β-fatty acid oxidation and de novo lipogenesis in cultured mouse hepatocytes provides a straight forward method to assess hepatocyte lipid handling. Here we describe a method for the isolation of primary mouse hepatocytes, and we demonstrate quantification of β-fatty acid oxidation and de novo lipogenesis, using radiolabeled substrates.
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The current study was undertaken to investigate he effects of human growth hormone(hGH) and porcine GH(pGH) on lipogenesis of adipose tissues from the rats and growing pigs. For the measurement of lipogenesis, epididymal fat tissue was taken from the rats after being killed and for the same purpose the backfat tissue was biopsyed from the pigs. Lipogenesis was determined by the amount of glucose converted to the total lipid. Initially, we investigated the effects of incubation time and glucose concentration on lipogenesis of the epididymal fat tissue of the rats for establishing the optimum condition. The results showed that lipogenesis was linearly increased as the incubation time increased (40, 80 and 120 min) and the total amount of lipogenesis was the same between 5mM and 20mM glucose in the KRB buffer. In in vitro study, adipose tissues were exposed to the varying concentrations of hGH during 48h culture; before the tissues were incubated in the KRB buffer for lipogenesis measurement. Human growth hormone added into the adipose tissue culture did not affect the lipogenesis of epididymal fat tissue of the rats. In in vivo study where the rats were injected daily with 200㎍ of hGH for 10 days, hGH added into the epididymal fat tissue culture did not affect the lipogenesis either. However, pGH as well as hGH contained in the culture media decreased the lipogenesis of the adipose tissue from growing pigs in dose-dependent manner. These results indicated that hGH decreased lipogenesis in the pigs, but not in the rats.
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The Spot 14 (S14; Thrsp) gene has been implicated in supporting regulated lipogenesis in mammals. S14 gene expression in liver is controlled by a wide variety of hormones and dietary factors in parallel with the major lipogenic enzyme genes. In addition, mice deleted for the S14 gene display reduced de novo lipogenesis in the lactating mammary gland. However, no decrease in hepatic lipogenesis was observed in the S14 null mouse. It was postulated that this difference could be due to the expression of a paralogous gene called S14R (S14 related; Mig12) in the liver but not mammary tissue. To test this hypothesis, we used small interfering RNA to simultaneously reduce levels of S14 and S14R in cultured primary hepatocytes. We found that rates of lipogenesis were decreased by approximately 65% in cells treated with insulin and high glucose. This reduction was associated with a decrease in total liver triacylglycerols and an altered morphology of lipid droplets. Expression of either S14 or S14R gene products was sufficient to fully restore normal lipogenesis. No change in the hepatic expression of other major lipogenic enzyme genes occurred during manipulation of S14 and/or S14R levels. These data support the hypothesis that both S14 and S14R are directly involved in supporting hepatic lipogenesis and that the two proteins play overlapping roles in this process.
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Lipid metabolism in liver is complex. In addition to importing and exporting lipid via lipoproteins, hepatocytes can oxidize lipid via fatty acid oxidation, or alternatively, synthesize new lipid via de novo lipogenesis. The net sum of these pathways is dictated by a number of factors, which in certain disease states leads to fatty liver disease. Excess hepatic lipid accumulation is associated with whole body insulin resistance and coronary heart disease. Tools to study lipid metabolism in hepatocytes are useful to understand the role of hepatic lipid metabolism in certain metabolic disorders. In the liver, hepatocytes regulate the breakdown and synthesis of fatty acids via β-fatty oxidation and de novo lipogenesis, respectively. Quantifying metabolism in these pathways provides insight into hepatic lipid handling. Unlike in vitro quantification, using primary hepatocytes, making measurements in vivo is technically challenging and resource intensive. Hence, quantifying β-fatty acid oxidation and de novo lipogenesis in cultured mouse hepatocytes provides a straight forward method to assess hepatocyte lipid handling. Here we describe a method for the isolation of primary mouse hepatocytes, and we demonstrate quantification of β-fatty acid oxidation and de novo lipogenesis, using radiolabeled substrates.
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