Yu 2013 Diabetes

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Yu L, Fink BD, Herlein JA, Sivitz WI (2013) Mitochondrial function in diabetes: novel methodology and new insight. Diabetes 62:1833-42.

» PMID: 23328129 Open Access

Yu L, Fink BD, Herlein JA, Sivitz WI (2013) Diabetes

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 ΔΨ, enabling more detailed study. We used a 2-deoxyglucose (2DOG) energy clamp to set ΔΨ at fixed levels and to quantify ATP production as 2DOG conversion to 2DOG-phosphate measured by one-dimensional (1)H and two-dimensional (1)H/(13)C heteronuclear single-quantum coherence nuclear magnetic resonance spectroscopy. These techniques proved far more sensitive than conventional (31)P nuclear magnetic resonance and allowed high-throughput study of small mitochondrial 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 ΔΨ 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.

Keywords: Diabetes, Heteronuclear single-quantum coherence nuclear magnetic resonance spectroscopy, Amplex Red, TPP electrode

O2k-Network Lab: US IA Iowa City Sivitz WI


Labels: MiParea: Respiration, Exercise physiology;nutrition;life style, mt-Medicine  Pathology: Diabetes  Stress:Oxidative stress;RONS  Organism: Rat  Tissue;cell: Skeletal muscle  Preparation: Isolated mitochondria 

Regulation: ATP, ATP production  Coupling state: LEAK, OXPHOS