Journal article
Mitochondrial Function in Diabetes: Novel Methodology and New Insight
Diabetes (New York, N.Y.), Vol.62(6), pp.1833-1842
06/01/2013
DOI: 10.2337/db12-1152
PMCID: PMC3661643
PMID: 23328129
Abstract
Interpreting mitochondrial function as affected by comparative physiologic conditions is confounding because individual functional parameters are interdependent. Here, we studied muscle mitochondrial function in insulin-deficient diabetes using a novel, highly sensitive, and specific method to quantify ATP production simultaneously with reactive oxygen species (ROS) at clamped levels of inner mitochondrial membrane potential (Delta Psi), enabling more detailed study. We used a 2-deoxyglucose (2DOG) energy clamp to set Delta Psi at fixed levels and to quantify ATP production as 2DOG conversion to 2DOG-phosphate measured by onedimensional H-1 and two-dimensional H-1/C-13 heteronuclear single quantum coherence nuclear magnetic resonance spectroscopy. These techniques proved far more sensitive than conventional P-31 nuclear magnetic resonance and allowed high-throughput study of small mitochonclrial isolates. Over conditions ranging from state 4 to state 3 respiration, ATP production was lower and ROS per unit of ATP generated was greater in mitochondria isolated from diabetic muscle. Moreover, ROS began to increase at a lower threshold for inner membrane potential in diabetic mitochondria. Further, ATP production in diabetic mitochondria is limited not only by respiration but also by limited capacity to use AT for ATP synthesis. In summary, we describe novel methodology for measuring ATP and provide new mechanistic insight into the dysregulation of ATP production and ROS in mitochondria of insulin-deficient rodents.
Details
- Title: Subtitle
- Mitochondrial Function in Diabetes: Novel Methodology and New Insight
- Creators
- Liping Yu - University of IowaBrian D. Fink - University of IowaJudith A. Herlein - University of IowaWilliam I. Sivitz - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Diabetes (New York, N.Y.), Vol.62(6), pp.1833-1842
- Publisher
- Amer Diabetes Assoc
- DOI
- 10.2337/db12-1152
- PMID
- 23328129
- PMCID
- PMC3661643
- ISSN
- 0012-1797
- eISSN
- 1939-327X
- Number of pages
- 10
- Grant note
- Veterans Affairs Medical Research Funds; US Department of Veterans Affairs Iowa Affiliate Fraternal Order of Eagles 5R01HL073166 / National Institutes of Health; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA R01HL073166 / NATIONAL HEART, LUNG, AND BLOOD INSTITUTE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Heart Lung & Blood Institute (NHLBI) I01BX000285 / Veterans Affairs; US Department of Veterans Affairs
- Language
- English
- Date published
- 06/01/2013
- Academic Unit
- Fraternal Order of Eagles Diabetes Research Center; Biochemistry and Molecular Biology; Medicine Administration; Endocrinology and Metabolism; Internal Medicine
- Record Identifier
- 9984627323702771
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