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Pyruvate Cycling Pathways and Glucose-Stimulated Insulin Secretion in Pancreatic Beta Cells.

Pyruvate Cycling Pathways and Glucose-Stimulated Insulin Secretion in Pancreatic Beta Cells.


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Pancreatic beta-cells secrete insulin in response to glucose. Intracellular glucose metabolism drives a cascade of events, including ATP production, calcium influx, and insulin processing, culminating in insulin granule exocytosis. However, insulin secretory mechanisms are incompletely understood. beta-cells have the capacity to flow pyruvate into the TCA cycle via the anaplerotic enzyme pyruvate carboxylase to engage one of several pathways of pyruvate recycling. Previous work demonstrated that pyruvate cycling was correlated with insulin secretion, and that NADPH may be involved in granule exocytosis. We hypothesized that NADPH-producing cytosolic enzymes isocitrate dehydrogenase (ICDc) and malic enzyme (MEc) may be involved in both pyruvate cycling and insulin secretion. ICDc expression was reduced using siRNA in the INS-1 derived cell line 832/13 and in isolated rat islets, which led decreased glucose-stimulated insulin secretion (GSIS), pyruvate cycling, and NADPH. Organic acid profiling revealed that decreased pyruvate cycling was compensated by an increase in lactate and stable pyruvate levels. This work established an important role for ICDc in maintaining GSIS through pyruvate-isocitrate cycling. MEc expression was reduced using siRNA in two beta-cell lines, 832/13 and 832/3, as well as isolated rat islets. MEc suppression inhibited GSIS in the 832/13 cells only, and these effects were not due to changes in pyruvate cycling, NADPH, or the organic acid profile. This suggests that in normal beta-cells, MEc does not participate in pyruvate cycling. Acetyl CoA carboxylase 1 (ACC1) is essential in de novo lipogenesis, which has been implicated in GSIS by other laboratories. Chronic, but not acute, inhibition of ACC1 via siRNA reduced insulin secretion independent of lipogenesis. ACC1 siRNA decreased glucose oxidation, pyruvate cycling, and ATP: ADP, due to an unexpected decrease in glucokinase protein. This work questions the use of ACC inhibitors in obesity and diabetes therapy. In summary, these studies on ICDc, MEc, and ACC1, coupled with concurrent work in our laboratory, eliminate two potential pyruvate cycling pathways (pyruvate-malate and pyruvate-citrate) and establish that pyruvate-isocitrate cycling is the critical pathway for control of GSIS. Future work will focus on identifying the signaling intermediate generated in the pyruvate-isocitrate pathway that links to insulin granule exocytosis


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Product Details
  • ISBN-13: 9781243434456
  • Publisher: Proquest, Umi Dissertation Publishing
  • Publisher Imprint: Proquest, Umi Dissertation Publishing
  • Height: 254 mm
  • Weight: 345 gr
  • ISBN-10: 1243434457
  • Publisher Date: 02 Sep 2011
  • Binding: Paperback
  • Spine Width: 11 mm
  • Width: 203 mm


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Pyruvate Cycling Pathways and Glucose-Stimulated Insulin Secretion in Pancreatic Beta Cells.
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Pyruvate Cycling Pathways and Glucose-Stimulated Insulin Secretion in Pancreatic Beta Cells.
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