Perry 2013 Diabetes
|Perry CG, Kane DA, Lanza IR, Neufer PD (2013) Methods for assessing mitochondrial function in diabetes. Diabetes 62:1041-53.|
Abstract: A growing body of research is investigating the potential contribution of mitochondrial function to the etiology of type 2 diabetes. Numerous in vitro, in situ, and in vivo methodologies are available to examine various aspects of mitochondrial function, each requiring an understanding of their principles, advantages, and limitations. This review provides investigators with a critical overview of the strengths, limitations and critical experimental parameters to consider when selecting and conducting studies on mitochondrial function. In vitro (isolated mitochondria) and in situ (permeabilized cells/tissue) approaches provide direct access to the mitochondria, allowing for study of mitochondrial bioenergetics and redox function under defined substrate conditions. Several experimental parameters must be tightly controlled, including assay media, temperature, oxygen concentration, and in the case of permeabilized skeletal muscle, the contractile state of the fibers. Recently developed technology now offers the opportunity to measure oxygen consumption in intact cultured cells. Magnetic resonance spectroscopy provides the most direct way of assessing mitochondrial function in vivo with interpretations based on specific modeling approaches. The continuing rapid evolution of these technologies offers new and exciting opportunities for deciphering the potential role of mitochondrial function in the etiology and treatment of diabetes.
- "High resolution designs (i.e., O2k, Oroboros Instruments) maximize respirometric sensitivity and precision (minimal O2 leak and highly sensitive electrodes), reducing the biological sample size required. Software advances in flux derivations of changes in chamber PO2 also permit real-time reporting of respiratory kinetics (Datlab, Oroboros Instruments), which improves data analyses over other systems requiring visual assessments of steady-state kinetics." (page 1042).
Labels: MiParea: Respiration, Instruments;methods, mt-Medicine Pathology: Diabetes
Organism: Human Tissue;cell: Skeletal muscle Preparation: Permeabilized tissue, Isolated mitochondria
Comparison of respirometric methods