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Assessing bioenergetic function in response to oxidative stress by metabolic profiling.
Brian P Dranka1, Gloria A Benavides, Anne R Diers
1Department of Pathology and Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Free Radical Biology & Medicine
|August 30, 2011
Summary
Mitochondria are key targets of oxidative stress in many diseases. Extracellular flux analysis offers a sensitive method to measure cellular bioenergetic responses to oxidative stress in various cell types.
Area of Science:
- Mitochondrial biology
- Cellular bioenergetics
- Oxidative stress research
Background:
- Mitochondria are crucial in cellular energy production and are vulnerable to oxidative stress, implicated in diseases like cardiovascular disease, diabetes, neurodegeneration, and cancer.
- Traditional methods for assessing mitochondrial function require substantial material and are limited in scope.
- Advancements in oxygen and pH measurement technologies enable the study of mitochondrial function in intact cells.
Purpose of the Study:
- To detail extracellular flux methods for assessing mitochondrial bioenergetic responses to oxidative stress.
- To demonstrate the application of these methods across diverse cell models, including renal, cardiovascular, nervous, and tumorigenic systems.
- To provide protocols for analyzing cellular bioenergetic function under oxidative stress conditions.
Main Methods:
- Utilizing high-resolution polarography and fluorescence techniques to measure oxygen concentration in solution.
- Employing extracellular flux analyzers to monitor real-time changes in oxygen concentration and pH in cultured adherent cells.
- Applying three distinct protocols to analyze the bioenergetic response of various cell types to oxidative stress.
Main Results:
- Extracellular flux methods provide a sensitive and efficient means to measure mitochondrial function in intact cells under oxidative stress.
- These methods are adaptable to a range of cell types, facilitating broad applicability in disease research.
- Demonstrated ability to analyze specific bioenergetic parameters in response to reactive species.
Conclusions:
- Extracellular flux analysis represents a significant advancement for studying mitochondrial dysfunction in oxidative stress-related pathologies.
- The described protocols enable comprehensive assessment of cellular bioenergetics in response to oxidative challenges.
- These methods enhance the capacity to investigate disease mechanisms and potential therapeutic targets involving mitochondrial health.
