Document Detail


Silencing of the mitochondrial NADH shuttle component aspartate-glutamate carrier AGC1/Aralar1 in INS-1E cells and rat islets.
MedLine Citation:
PMID:  19764902     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Transfer of reducing equivalents between cytosolic compartments and the mitochondrial matrix is mediated by NADH shuttles. Among these, the malate-aspartate shuttle has been proposed to play a major role in beta-cells for the control of glucose-stimulated insulin secretion. AGC1 or Aralar1 (aspartate-glutamate carrier 1) is a key component of the malate-aspartate shuttle. Overexpression of AGC1 increases the capacity of the malate-aspartate shuttle, resulting in enhanced metabolism-secretion coupling, both in INS-1E cells and rat islets. In the present study, knockdown of AGC1 was achieved in the same beta-cell models, using adenovirus-mediated delivery of shRNA (small-hairpin RNA). Compared with control INS-1E cells, down-regulation of AGC1 blunted NADH formation (-57%; P<0.05), increased lactate production (+16%; P<0.001) and inhibited glucose oxidation (-22%; P<0.01). This correlated with a reduced secretory response at 15 mM glucose (-25%; P<0.05), while insulin release was unchanged at intermediate 7.5 mM and basal 2.5 mM glucose. In isolated rat islets, efficient AGC1 knockdown did not alter insulin exocytosis evoked by 16.7 mM glucose. However, 4 mM amino-oxyacetate, commonly used to block transaminases of the malate-aspartate shuttle, inhibited glucose-stimulated insulin secretion to similar extents in INS-1E cells (-66%; P<0.01) and rat islets (-56%; P<0.01). These results show that down-regulation of the key component of the malate-aspartate shuttle AGC1 reduced glucose-induced oxidative metabolism and insulin secretion in INS-1E cells, whereas similar AGC1 knockdown in rat islets did not affect their secretory response.
Authors:
Marina Casimir; Blanca Rubi; Francesca Frigerio; Gaelle Chaffard; Pierre Maechler
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-12-10
Journal Detail:
Title:  The Biochemical journal     Volume:  424     ISSN:  1470-8728     ISO Abbreviation:  Biochem. J.     Publication Date:  2009 Dec 
Date Detail:
Created Date:  2009-11-25     Completed Date:  2010-01-14     Revised Date:  2011-11-09    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  England    
Other Details:
Languages:  eng     Pagination:  459-66     Citation Subset:  IM    
Affiliation:
Department of Cell Physiology and Metabolism, Faculty of Medicine, University of Geneva, 1 rue Michel-Servet, 1211 Geneva 4, Switzerland.
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MeSH Terms
Descriptor/Qualifier:
Adenosine Triphosphate / metabolism
Aminooxyacetic Acid / pharmacology
Animals
Calcium / metabolism
Cell Line, Tumor
Enzyme Inhibitors / pharmacology
Glucose / metabolism,  pharmacology
Glutamic Acid / metabolism
Insulin / secretion
Islets of Langerhans / cytology,  drug effects,  metabolism*
Lactates / metabolism
Membrane Potential, Mitochondrial / drug effects
Membrane Transport Proteins / genetics,  metabolism*
Mitochondria / metabolism,  physiology
Mitochondrial Proteins / genetics,  metabolism*
RNA Interference*
Rats
Chemical
Reg. No./Substance:
0/Enzyme Inhibitors; 0/Lactates; 0/Membrane Transport Proteins; 0/Mitochondrial Proteins; 0/Slc25a12 protein, rat; 11061-68-0/Insulin; 50-99-7/Glucose; 56-65-5/Adenosine Triphosphate; 56-86-0/Glutamic Acid; 645-88-5/Aminooxyacetic Acid; 7440-70-2/Calcium

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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