Description
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. Neuromethods Preface… Preface… Table of Contents… Contributing Authors… 1. Three-Dimensional Reconstruction of Neuronal Mitochondria by Electron Tomography Guy Perkins 2. Measuring Mitochondrial Shape with ImageJ Ronald A. Merrill, Kyle H. Flippo, and Stefan Strack 3. Live Imaging Mitochondrial Transport in Neurons Meredith M. Course, Chung-Han Hsieh, Pei-I Tsai, Jennifer A. Codding-Bui, Atossa Shaltouki, and Xinnan Wang 4. Techniques to Investigate Bioenergetics of Mitochondria William I. Sivitz 5. Respirometry in Neurons Liang Zhang and Eugenia Trushina 6. Measuring ATP in Axons with FRET Lauren Y. Shields, Bryce A. Mendelsohn, and Ken Nakamura 7. Characterizing Metabolic States using Fluorescence Lifetime Imaging Microscopy (FLIM) of NAD(P)H Thomas S. Blacker and Michael R. Duchen 8. Techniques for Simultaneous Mitochondrial and Cytosolic Ca2+ Imaging in Neurons Jacob E. Rysted, Zhihong Lin, and Yuriy M. Usachev 9. Live Imaging of Mitochondrial ROS Production and Dynamic Redox Balance in Neurons Karolina Can, Sebastian Kgler, and Michael Mller 10. Monitoring of Permeability Transition Pore Openings in Isolated Individual Brain Mitochondria Nickolay Brustovetsky and Tatiana Brustovetsky 11. Examination of Mitochondrial Ion Conductance by Patch Clamp in Intact Neurons and Mitochondrial Membrane Preparations Elizabeth A. Jonas and Nelli Mnatsakanyan 12. Monitoring of Permeability Transition Pore Openings in Mitochondria of Cultured Neurons Tatiana Brustovetsky and Nickolay Brustovetsky 13. Protocols for Assessing Mitophagy in Neuronal Cell Lines and Primary Neurons Ruben K. Dagda and Monica Rice 14. Examining Mitochondrial Function at Synapses In Situ Gregory T. Macleod and Maxim V. Ivannikov 15. Proteomic Analysis of Neuronal Mitochondria Kelly L. Stauch and Howard S. Fox 16. Measuring Mitochondrial Pyruvate Oxidation Lawrence R. Gray, Alix A.J. Rouault, Lalita Oonthonpan, Adam J. Rauckhorst, Julien A. Sebag, and Eric B. Taylor
