Mitochondrial Bioenergetics Methods and Protocols

This updated volume provides a concise set of protocols on the conceptual principles and practical considerations pertinent to research in mitochondrial function. Driven by the impact of drug- and environmental chemical-induced disturbances on mitochondrial function, hereditary deficiencies, and the age-related decline in bioenergetic performance, this book explores the latest approaches developed in this field by presenting practical principles and assays for quantitative assessments of various parameters reflecting aspects of mitochondrial bioenergetics. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and up-to-date, Mitochondrial Bioenergetics: Methods and Protocols, Third Edition serves as an ideal guide for scientists endeavoring to enhance the quality of mitochondrial research.

Evaluation of Respiration with a Clark-Type Electrode in Isolated Mitochondria, Intact and Permeabilized Cells, and Explants from Animal Tissues.- High-Resolution Respirometry Methodology for Bioenergetic and Metabolic Studies in Intact Brain Slices.- Plate-Based Measurement of Respiration by Isolated Mitochondria and by Intact Cells.- Metabolic Phenotyping of Synaptic Mitochondria Using MitoPlatesTM and Synaptoneurosomes.- Characterization of the Mitochondria Function and Metabolism in Skin Fibroblasts Using the Biolog MitoPlate S-1.- Measurement of Mitochondrial ROS Formation.- Mitochondrial Membrane Potential (??) Fluctuations Associated with the Metabolic States of Mitochondria.- The Relation between Mitochondrial Membrane Potential and Reactive Oxygen Species Formation.- Creating Physiological Cell Environments In Vitro: Adjusting Cell Culture Media Composition and Oxygen Levels to Investigate Mitochondrial Function and Cancer Metabolism.- Imaging Mitochondrial Axonal Transport in Human Induced Pluripotent Stem Cell-Derived Neurons.- Understanding Mitochondrial and Lysosomal Dynamics by Fluorescent Microscopy.- Mitochondrial Morphofunctional Profiling in Primary Human Skin Fibroblasts Using TMRM and Mitotracker Green Co-Staining.- Absolute Quantification of Cellular and Cell-Free Mitochondrial DNA Copy Number from Human Blood and Urinary Samples Using Real Time Quantitative PCR.- Isolation and Analysis of Mitochondrial Small RNAs from Rat Liver Tissue and HepG2 Cells.- Methodologies for Mitochondrial Omic Profiling during Spaceflight.