MEASURING MITOCHONDRIAL BIOENERGETICS IN INS-1E INSULINOMA CELLS

被引：39
|
作者：
Affourtit, Charles [1 ]
Brand, Martin D. [1 ,2 ]
机构：
[1] MRC, Dunn Human Nutr Unit, Cambridge, England
[2] Buck Inst Age Res, Novato, CA USA
来源：
METHODS IN ENZYMOLOGY, VOL 457: MITOCHONDRIAL FUNCTION, PARTB MITOCHONDRIAL PROTEIN KINASES, PROTEIN PHOSPHATASES AND MITOCHONDRIAL DISEASES | 2009年 / 457卷
基金：
英国医学研究理事会;
关键词：
DOWN ELASTICITY ANALYSIS; PROTON LEAK; UNCOUPLING PROTEIN-2; RESPIRATION RATE; OXIDATIVE-PHOSPHORYLATION; LIVER-MITOCHONDRIA; ENERGY-METABOLISM; SKELETAL-MUSCLE; INNER MEMBRANE; HEPATOCYTES;
D O I：
10.1016/S0076-6879(09)05023-X
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
引用
收藏
页码：405 / 424
页数：20
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