Document Detail


Measuring mitochondrial bioenergetics in INS-1E insulinoma cells.
MedLine Citation:
PMID:  19426881     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Pancreatic beta cells secrete insulin in response to raised blood glucose levels. This glucose-stimulated insulin secretion (GSIS) depends on mitochondrial function and is regulated by the efficiency with which oxidative metabolism is coupled to ATP synthesis. Uncoupling protein-2 (UCP2) affects this coupling efficiency and is therefore a plausible pathological and physiological regulator of GSIS. In this respect, it is important to be able to measure coupling efficiencies accurately. Here, we describe experimental protocols to determine the coupling efficiency of trypsinized INS-1E cells, a popular beta cell model, and we present practical details of our RNA interference studies to probe the effect of UCP2 knockdown on this efficiency. We also introduce a method to determine coupling efficiencies noninvasively in attached cells and discuss theoretical and practical aspects of a modular-kinetic approach to describe and understand cellular bioenergetics.
Authors:
Charles Affourtit; Martin D Brand
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Methods in enzymology     Volume:  457     ISSN:  1557-7988     ISO Abbreviation:  Meth. Enzymol.     Publication Date:  2009  
Date Detail:
Created Date:  2009-05-11     Completed Date:  2009-07-13     Revised Date:  2014-02-19    
Medline Journal Info:
Nlm Unique ID:  0212271     Medline TA:  Methods Enzymol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  405-24     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Line, Tumor
Energy Metabolism
Gene Knockdown Techniques
Humans
Insulin-Secreting Cells / chemistry,  metabolism
Insulinoma / genetics,  metabolism*
Ion Channels / analysis,  genetics*,  metabolism*
Kinetics
Mitochondria / chemistry,  metabolism*
Mitochondrial Proteins / analysis,  genetics*,  metabolism*
Oxygen Consumption*
RNA Interference
Grant Support
ID/Acronym/Agency:
MC_U105663137//Medical Research Council; //Medical Research Council
Chemical
Reg. No./Substance:
0/Ion Channels; 0/Mitochondrial Proteins; 0/mitochondrial uncoupling protein 2

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