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 Compartments1. Isolation of mitochondria from animal cells and yeastFrancesco Pallotti and Liza Pon2. Isolation and characterization of mitochondria-associated ER membranesEstela Area-Gomez3. Import of proteins into mitochondriaNikolaus Pfanner and Thomas Becker4. Proximity labeling methods to assess protein-protein interactions in yeast mitochondriaJie-Ning Yang and Liza Pon Part 2 - Assays for Mitochondrial Metabolic Activities5. Biochemical assays of TCA cycle and β-oxidation metabolitesMichael J. Bennett, Sheng Feng and Ann Saada (Reisch)6. Biochemical assays of respiratory chain complex activity Robert Taylor7. Assessing mitochondrial bioenergetics in whole cells and isolated organelles by microplate respirometry and polarographyErich Gnaiger and Orian Shirihai8. Analyzing electron transport chain supercomplexesJose Antonio Enriquez9. Assays of mitochondrial ATP synthesis in animal cells and tissuesPaolo Pinton and Hiromi Imamura10. Visualization of mitochondrial membrane potential in mammalian cellsNoemi Esteras, Merel Adjobo-Hermans, Andray Abramov and Werner JH Koopman11. Visualization of mitochondrial function in living cells using biosensors (SoNAR, ro-GFP, DiOC6/Tom70 mCherry; DHE; Mitosox)Yi Yang and Liza PonPart 3 - Assays for Other Mitochondrial Activities12. Histochemical methods to measure respiratory chain activity and visualize mitochondria and mitochondrial gene products Kurenai Tanji and Sandra Franco-Iborra13. Analysis of mitochondrial phospholipids and cardiolipinMichael Schlame and Estela Area-Gomez14. Biosensors for Detection of Calcium and pHDiego Destefani15. Measurement of membrane permeability and the mitochondrial permeability transition Paolo Bernardi and Michela CarraroPart 4 - Mitochondrial Genes and Gene Expression 16. Detection of mtDNA mutationsAli Naini, Sara Shanske, Robert Gilkerson and Jiuhong Pang17. Visualization of mitochondrial DNA replicationAgnel Sfeir18. Cybrid technology Carlos Moraes and Sandra Bacman19. Manipulation of mitochondrial genes and mtDNA heteroplasmyCarlos Moraes, Sandra Bacman and Michal MinczukPart 5 - Assays for Mitochondrial Dynamics20. Assays for mitochondrial fusion and fission Laura Lackner, Jason Lee, Jason Mears and Suzanne Hoppins21. Methods to visualize mitochondria and mitochondrial motilityLiza Pon, Theresa Swayne, Ryo Higuchi-Sanabria and Pin-Chao Liao22. In vitro and in vivo detection of mitophagy in animal cells, mice, and yeast (Keima; red/green mitophagosomes)AppendicesA. Basic properties of mitochondriaLuis García-RodríguezB. Inhibitors of the respiratory chain and of ATP synthesisNanette Orme-Johnson and Eric A. SchonC. Linearized maps of selected mitochondrial genomesEric SchonD. Mitochondrial genetic codes in various organisms Eric SchonE. Gene products present in mitochondria of yeast and animal cells Eric Schon