Main 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.

Contents:

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 Kugler, and Michael Muller

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