alexa Comparative Analysis of the Mitochondrial Physiology of
ISSN: 2167-7662

Bioenergetics: Open Access
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Research Article

Comparative Analysis of the Mitochondrial Physiology of Pancreatic b Cells

Chul Kim1, Pinal Patel1, Lindsey M. Gouvin2, Melissa L. Brown1, Ahmed Khalil3, Elizabeth M Henchey1, Alejandro P. Heuck2 and Nagendra Yadava1,3,4*
1Pioneer Valley Life Sciences Institute, Springfield, MA, USA
2Departments of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, MA,USA
3Department of Biology, University of Massachusetts, Amherst, MA, USA
4Division of Endocrinology, Diabetes & Metabolism at Baystate Medical Center of Tufts University School of Medicine, Springfield, MA, USA
Corresponding Author : Nagendra Yadava
Assistant Professor, John Adams Investigator
Pioneer Valley Life Sciences Institute
Springfield, 01107 MA, USA
Tel: 413-794-0786
Fax: 413-794-0857
E-mail: [email protected]
Received November 26, 2013; Accepted January 15, 2014; Published January 15, 2014
Citation: Kim C, Patel P, Gouvin LM, Brown ML, Khalil A, et al. (2014) Comparative Analysis of the Mitochondrial Physiology of Pancreatic b Cells. Bioenergetics 3:110. doi:10.4172/2167-7662.1000110
Copyright: © 2014 Kim C, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
 

Abstract

The mitochondrial metabolism of b cells is thought to be highly specialized. Its direct comparison with other cells using isolated mitochondria is limited by the availability of islets/b cells in sufficient quantity. In this study, we have compared mitochondrial metabolism of INS1E/b cells with other cells in intact and permeabilized states. To selectively permeabilize the plasma membrane, we have evaluated the use of perfringolysin O (PFO) in conjunction with microplate-based respirometry. PFO is a protein that binds membranes based on a threshold level of active cholesterol. Therefore, unless active cholesterol reaches a threshold level in mitochondria, they are expected to remain untouched by PFO. Cytochrome c sensitivity tests showed that in PFO-permeabilized cells, the mitochondrial integrity is completely preserved. Our data show that a time-dependent decline of the oligomycin-insensitive respiration observed in INS1E cells was due to a limitation in substrate supply to the respiratory chain. We predict that it is linked with the b cellspecific metabolism involving metabolites shuttling between the cytoplasm and mitochondria. In permeabilized b cells, the Complex I-dependent respiration was either transient or absent because of the inefficient TCA cycle. The TCA cycle insufficiency was confirmed by analysis of the CO2 evolution. This may be linked with lower levels of NAD+, which is required as a co-factor for CO2 producing reactions of the TCA cycle. b cells showed comparable Ox Phos and respiratory capacities that were not affected by the inorganic phosphate (Pi) levels in the respiration medium. They showed lower ADP-stimulation of the respiration on different substrates. We believe that this study will significantly enhance our understanding of the b cell mitochondrial metabolism.

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