Difference between revisions of "Yu 2013 Diabetes"
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|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. | |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 | |keywords=Diabetes, Heteronuclear single-quantum coherence nuclear magnetic resonance spectroscopy, Amplex Red, TPP electrode | ||
|mipnetlab=US IA Iowa City Sivitz WI | |||
}} | }} | ||
{{Labeling | {{Labeling | ||
|area=Respiration, Exercise physiology;nutrition;life style, mt-Medicine | |area=Respiration, Exercise physiology;nutrition;life style, mt-Medicine | ||
|diseases=Diabetes | |||
|injuries=Oxidative stress;RONS | |||
|organism=Rat | |organism=Rat | ||
|tissues=Skeletal muscle | |tissues=Skeletal muscle | ||
|preparations=Isolated mitochondria | |preparations=Isolated mitochondria | ||
|topics=ATP, ATP production | |topics=ATP, ATP production | ||
|couplingstates=LEAK, OXPHOS | |couplingstates=LEAK, OXPHOS | ||
}} | }} | ||
Latest revision as of 14:24, 28 March 2018
| Yu L, Fink BD, Herlein JA, Sivitz WI (2013) Mitochondrial function in diabetes: novel methodology and new insight. Diabetes 62:1833-42. |
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
