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Mitochondria Biology

1st Edition - March 13, 2020

Editors: Liza A. Pon, Eric A. Schon

Hardback ISBN:

9 7 8 - 0 - 1 2 - 8 2 0 2 2 8 - 9

eBook ISBN:

9 7 8 - 0 - 1 2 - 8 2 0 2 2 9 - 6

Methods in Cell Biology Volume 155 provides an update on the step-by-step "how-to" methods to study mitochondrial structure, function and biogenesis contained in the first two… Read more

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Methods in Cell Biology Volume 155 provides an update on the step-by-step "how-to" methods to study mitochondrial structure, function and biogenesis contained in the first two editions. As in the previous editions, biochemical, cell biological, and genetic approaches are presented along with sample results, interpretations, and pitfalls for each method. New chapters in this update include Isolation of Mitochondria and Analysis of Mitochondrial Compartments, Isolation of Mitochondria from Animal Cells and Yeast, Isolation and Characterization of Mitochondria-Associated ER Membranes, Import of Proteins into Mitochondria, Proximity Labeling Methods to Assess Protein-Protein Interactions in Yeast Mitochondria, and more.

Part 1 - Isolation of Mitochondria and Analysis of Mitochondrial Compartments

1. Isolation of mitochondria from animal cells and yeast
Francesco Pallotti and Liza Pon

2. Isolation and characterization of mitochondria-associated ER membranes
Estela Area-Gomez

3. Import of proteins into mitochondria
Nikolaus Pfanner and Thomas Becker

4. Proximity labeling methods to assess protein-protein interactions in yeast mitochondria
Jie-Ning Yang and Liza Pon

Part 2 - Assays for Mitochondrial Metabolic Activities

5. Biochemical assays of TCA cycle and β-oxidation metabolites
Michael J. Bennett, Sheng Feng and Ann Saada (Reisch)

6. Biochemical assays of respiratory chain complex activity
Robert Taylor

7. Assessing mitochondrial bioenergetics in whole cells and isolated organelles by microplate respirometry and polarography
Erich Gnaiger and Orian Shirihai

8. Analyzing electron transport chain supercomplexes
Jose Antonio Enriquez

9. Assays of mitochondrial ATP synthesis in animal cells and tissues
Paolo Pinton and Hiromi Imamura

10. Visualization of mitochondrial membrane potential in mammalian cells
Noemi Esteras, Merel Adjobo-Hermans, Andray Abramov and Werner JH Koopman

11. Visualization of mitochondrial function in living cells using biosensors (SoNAR, ro-GFP, DiOC6/Tom70 mCherry; DHE; Mitosox)
Yi Yang and Liza Pon

Part 3 - Assays for Other Mitochondrial Activities

12. Histochemical methods to measure respiratory chain activity and visualize mitochondria and mitochondrial gene products
Kurenai Tanji and Sandra Franco-Iborra

13. Analysis of mitochondrial phospholipids and cardiolipin
Michael Schlame and Estela Area-Gomez

14. Biosensors for Detection of Calcium and pH
Diego Destefani

15. Measurement of membrane permeability and the mitochondrial permeability transition
Paolo Bernardi and Michela Carraro

Part 4 - Mitochondrial Genes and Gene Expression

16. Detection of mtDNA mutations
Ali Naini, Sara Shanske, Robert Gilkerson and Jiuhong Pang

17. Visualization of mitochondrial DNA replication
Agnel Sfeir

18. Cybrid technology
Carlos Moraes and Sandra Bacman

19. Manipulation of mitochondrial genes and mtDNA heteroplasmy
Carlos Moraes, Sandra Bacman and Michal Minczuk

Part 5 - Assays for Mitochondrial Dynamics

20. Assays for mitochondrial fusion and fission
Laura Lackner, Jason Lee, Jason Mears and Suzanne Hoppins

21. Methods to visualize mitochondria and mitochondrial motility
Liza Pon, Theresa Swayne, Ryo Higuchi-Sanabria and Pin-Chao Liao

22. In vitro and in vivo detection of mitophagy in animal cells, mice, and yeast (Keima; red/green mitophagosomes)

Appendices
A. Basic properties of mitochondria
Luis García-Rodríguez
B. Inhibitors of the respiratory chain and of ATP synthesis
Nanette Orme-Johnson and Eric A. Schon
C. Linearized maps of selected mitochondrial genomes
Eric Schon
D. Mitochondrial genetic codes in various organisms
Eric Schon
E. Gene products present in mitochondria of yeast and animal cells
Eric Schon

Liza A. Pon

Liza A. Pon, PhD, is a Professor of Pathology and Cell Biology and the Institute of Human Nutrition, and Director of the Confocal and Specialized Microscopy Shared Resource at Columbia University. As a PhD student at Tufts University, she carried out the first characterization of a protein now known as StaR, steroidogenic acute regulatory protein, which mediates transfer of cholesterol within mitochondria during steroid hormone biosynthesis. As a post-doctoral scientist with Gottfried (Jeff) Schatz at the University of Basel, she studied the mechanism of import of proteins into mitochondria. Dr. Pon joined Columbia University in 1990 and has been there ever since. Her research focuses on mitochondrial motility, dynamics, quality control and interactions with other organelles, and how these processes affect cell cycle progression, cellular fitness and lifespan. Recent studies in the laboratory focus on mechanisms for detection and removal of unfolded and damaged proteins not just in mitochondria but also in the ER in models of aging and disease.

Affiliations and expertise

Department of Pathology and Cell Biology and Institute of Human Nutrition, Columbia University

Eric A. Schon

Eric A. Schon, PhD, is the Lewis P. Rowland Professor of Neurology (in Genetics and Development) at Columbia University. After graduating Columbia University with a B.S. in Chemical Engineering, he spent 10 years as a Technical Brand Manager for the Procter & Gamble Company in Cincinnati, OH. After leaving P&G, he received a PhD in Biological Chemistry from the University of Cincinnati, followed by a postdoctoral fellowship at the Harvard Medical School. He moved to Columbia as an Associate Research Scientist in 1984, and has been there ever since. Dr. Schon's laboratory studies the molecular genetics of neurological and neuromuscular diseases, with particular focus on mitochondrial disorders. The research has two principal goals: (1) to use the tools of molecular and cell biology in order to gain insight into the etiology, pathogenesis, and treatment of these devastating diseases, and (2) to build on this knowledge in order to ask more fundamental biological questions relating to nuclear-mitochondrial communication, mitochondrial biogenesis, and mtDNA plasticity. Most recently, the laboratory has become interested in understanding the structural and functional relationships between mitochondria and the endoplasmic reticulum in the pathogenesis of Alzheimer disease.

Affiliations and expertise

Department of Neurology (in Genetics and Development), Columbia University