Assessing Bioenergetic Function in Response to Oxidative Stress by Metabolic Profiling

Overview

Journal Free Radic Biol Med

Specialties Biochemistry
Biology
General Medicine

Date 2011 Aug 30

PMID 21872656

Citations 262

Authors

Brian P Dranka

Gloria A Benavides

Anne R Diers

Samantha Giordano

Blake R Zelickson

Colin Reily

Luyun Zou

John C Chatham

Bradford G Hill

Jianhua Zhang

Aimee Landar

Victor M Darley-Usmar

Affiliations

Soon will be listed here.

Abstract

It is now clear that mitochondria are an important target for oxidative stress in a broad range of pathologies, including cardiovascular disease, diabetes, neurodegeneration, and cancer. Methods for assessing the impact of reactive species on isolated mitochondria are well established but constrained by the need for large amounts of material to prepare intact mitochondria for polarographic measurements. With the availability of high-resolution polarography and fluorescence techniques for the measurement of oxygen concentration in solution, measurements of mitochondrial function in intact cells can be made. Recently, the development of extracellular flux methods to monitor changes in oxygen concentration and pH in cultures of adherent cells in multiple-sample wells simultaneously has greatly enhanced the ability to measure bioenergetic function in response to oxidative stress. Here we describe these methods in detail using representative cell types from renal, cardiovascular, nervous, and tumorigenic model systems while illustrating the application of three protocols to analyze the bioenergetic response of cells to oxidative stress.

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