Mitochondria

Mitochondria

2nd Edition - April 10, 2007

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  • Editors: Liza Pon, Eric Schon
  • eBook ISBN: 9780080489377

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Description

This book provides an update on the step-by-step "how to" methods for the study mitochondrial structure, function, and biogenesis contained in the successful first edition. As in the previous edition, the biochemical, cell biological, and genetic approaches are presented along with sample results, interpretations, and pitfalls from each method.

Readership

Researchers and graduate students in cell biology, developmental biology, and genetics.
Academic and research libraries, pharmaceutical and biotechnology companies, and private research facilities.

Table of Contents

  • Isolation and Subfractionation of Mitochondria
    1. Isolation and Subfractionation of Mitochondria in Animal Cells
    2. Purification and Subfractionation of Mitochondria from the Yeast Saccharomyces cerevisiae
    3. Isolation and Subfractionation of Mitochondria from Plants

    Biochemical Assays of Mitochondrial Activity
    4. Biochemical Assays of Respiratory Chain Complex Activity
    5. Polarographic Assays of Respiratory Chain Complex Activity
    6. Optical Imaging Techniques (Histochemical, Immunohistochemical and In Situ Hybridization Staining Methods) to Visualize Mitochondria
    7. Assay of Mitochondrial ATP Synthesis in Animal Cells and Tissues
    8. Measurement of the Ratio of Lactate to Pyruvate in Skin Fibroblast Cultures
    9. Assays of Fatty Acid ƒÒ-Oxidation Activity
    10. Biochemical Assays for Mitochondrial Activity: Assays of TCA Cycle Enzymes and PDHc
    11. Assays of Cardiolipin Levels
    12. Measurement of VDAC Permeability in Intact Mitochondria and in Reconstituted Systems
    13. Methods for Studying Fe/S Cluster Protein Maturation in Yeast Mitochondria

    Assays for Mitochondrial Respiratory Activity and Permeability in Living Cells
    14. Imaging of Mitochondrial Polarization and Depolarization with Cationic Fluorophores
    15. Biosensors for the Detection of Calcium and pH
    16. Measurement of Membrane Permeability and the Permeability Transition of Mitochondria
    17. Luciferase Expression for ATP Imaging: Application to Cardiac Myocytes

    Oxidative Stress Measurements
    18. Measurement of Reactive Oxygen Species in Cells and Mitochondria
    19. Measurements of the Antioxidant Enzyme Activities of Superoxide Dismutase, Catalase, and Glutathione Peroxidase
    20. Methods for Measuring the Regulation of Respiration by Nitric Oxide
    21. Methods for Determining the Modification of Protein Thiols by Reactive Lipids

    Mitochondrial Genes and Gene Expression
    22. Detection of Mutations in mtDNA
    23. Diagnostic Assays for Defects in mtDNA Replication and Transcription in Yeast and Humans
    24. Microdissection and Analytical PCR for the Investigation of mtDNA Lesions
    25. Transmitochondrial Technology in Animal Cells
    26. Genetic Transformation of Saccharomyces cerevisiae and Chlamydomonas Reinhardtii Mitochondria
    27. Generation of Transmitochondrial Mice: Development of Xenomitochondrial Mice to Model Neurodegenerative Diseases
    28. In vivo and In organello Analysis of Mitochondrial Translation
    Assays for Mitochondrial Morphology and Motility
    29. Visualization of Mitochondria in Budding Yeast
    30. Visualization and Quantification of Mitochondrial Dynamics in Living Animal Cells
    31. Cell-Free Assays for Mitochodnrial-Cytoskeletal Interactions
    32. Assays to Measure Mitochondrial Fusion In Vitro and the GTPase Activity of Mitochondrial Fission Proteins

    Methods to Determine Protein Localization to Mitochondria
    33. Electrophoretic Methods to Isolate Protein Complexes from Mitochondria
    34. Analysis of Protein-Protein Interactions in Mitochondria
    35. Analysis and Prediction of Mitochondrial Targeting Signals
    36. Import of Proteins into Mitochondria

    Appendices
    Appendix 1: Basic Properties of Mitochondria
    Appendix 2: Direct and Indirect Inhibitors of Mitochondrial ATP Synthesis
    Appendix 3: Linearized Maps of Mitochondrial Genomes from Representative Organisms
    Appendix 4: Mitochondrial Genetic Codes in Various Organisms
    Appendix 5: Gene Products Present in Mitochondria of Yeast and Animal Cells
    Appendix 6: Changes in the Mitochondrial Transcriptome and Proteome Under Various Stresses and Growth Conditions

Product details

  • No. of pages: 944
  • Language: English
  • Copyright: © Academic Press 2007
  • Published: April 10, 2007
  • Imprint: Academic Press
  • eBook ISBN: 9780080489377

About the Editors

Liza 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 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

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