Mitochondria Biology

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. 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.

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 ß-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