XV, 344 p. 74 illus., 51 illus. in color. online resource
This volume describes a broad spectrum of experimental approaches for investigating structure, function, and transport of neuronal mitochondria in health and disease. Most of these approaches were only recently developed and range from electron tomography-based 3D reconstruction of mitochondrial cristae to patch clamp recording from mitochondria in intact neurons. The chapters in this book cover topics such as mitochondrial proteomics, fluorescence lifetime imaging, respirometry and mitophagy, as well as optical approaches based on the use of genetically engineered fluorescent sensors for monitoring synaptic ATP and axonal ROS generation, mitochondrial Ca2+ cycling and pH changes, and mitochondrial dynamics and axonal trafficking in live neurons. Each chapter also discusses difficulties, tips, tricks, and precautions to take. Neuromethods series style chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Cutting-edge and comprehensive, Techniques to Investigate Mitochondrial Function in Neurons is a valuable and useful resource for a broad range of investigators interested in the function of neuronal mitochondria in health and disease states.
Three-Dimensional Reconstruction of Neuronal Mitochondria by Electron Tomography -- Measuring Mitochondrial Shape with ImageJ -- Live Imaging Mitochondrial Transport in Neurons -- Techniques to Investigate Bioenergetics of Mitochondria -- Respirometry in Neurons -- Measuring ATP in Axons with FRET -- Characterizing Metabolic States using Fluorescence Lifetime Imaging Microscopy (FLIM) of NAD(P)H -- Techniques for Simultaneous Mitochondrial and Cytosolic Ca2+ Imaging in Neurons -- Live Imaging of Mitochondrial ROS Production and Dynamic Redox Balance in Neurons -- Monitoring of Permeability Transition Pore Openings in Isolated Individual Brain Mitochondria -- Examination of Mitochondrial Ion Conductance by Patch Clamp in Intact Neurons and Mitochondrial Membrane Preparations -- Monitoring of Permeability Transition Pore Openings in Mitochondria of Cultured Neurons -- Protocols for Assessing Mitophagy in Neuronal Cell Lines and Primary Neurons -- Examining Mitochondrial Function at Synapses In Situ -- Proteomic Analysis of Neuronal Mitochondria -- Measuring Mitochondrial Pyruvate Oxidation