Mitochondrial function. Volume 547 /

Detalles Bibliográficos
Clasificación:Libro Electrónico
Otros Autores: Murphy, Anne N. (Anne Neville) (Editor ), Chan, David C., 1966- (Editor )
Formato: Electrónico eBook
Idioma:Inglés
Publicado: Amsterdam : Academic Press, 2014.
Colección:Methods in enzymology ; v. 547.
Temas:
Mitochondrial pathology.
Mitochondrial DNA.
Neurology > Research.
DNA, Mitochondrial
Maladies mitochondriales.
ADN mitochondrial.
Neurologie > Recherche.
SCIENCE > Life Sciences > Anatomy & Physiology.
Mitochondrial DNA
Mitochondrial pathology
Neurology > Research
Internet Resources.
Acceso en línea:Texto completo
Texto completo
Tabla de Contenidos:
  • 1.High-Content Functional Genomic Screening to Identify Novel Regulators of the PINK1
  • Parkin Pathway / Robert A. Screaton
  • 1.Introduction and Theory
  • 2.General Screen Design Strategy
  • 3.RNAi Screen for Genes Involved Required for PARK2 Translocation
  • 4.Equipment
  • 5.Materials
  • 6.Genome Screen Protocol
  • Acknowledgments
  • References
  • 2.Measurement of Mitochondrial Turnover and Life Cycle Using MitoTimer / Orian S. Shirihai
  • 1.Introduction
  • 2.Expressing MitoTimer in Cells
  • 3.Imaging MitoTimer
  • 4.Interpreting MitoTimer Readout
  • 5.Summary
  • References
  • 3.Monitoring Mitophagy in Mammalian Cells / Quan Chen
  • 1.Introduction
  • 2.Transmission Electron Microscopy
  • 3.Western Blot Analysis of Mitophagy
  • 4.Fluorescence Methods for Analyzing Mitophagy
  • 5.Analyzing Mitophagy Using Mitochondria Mass
  • 6.Mitochondria-Targeting Probes for Mitophagy Assay
  • 7.Mitophagy Inducer and Inhibitors
  • 8.Future Perspectives
  • Acknowledgments
  • References
  • 4.Photoactivatable Green Fluorescent Protein-Based Visualization and Quantification of Mitochondrial Fusion and Mitochondrial Network Complexity in Living Cells / Brian A. Roelofs
  • 1.Introduction
  • 2.Visualization and Quantification of Mitochondrial Fusion in Living Cells
  • 3.Estimation of Relative Size of Mitochondrial Units and Mitochondrial Network Complexity
  • 4.Time-Lapse Imaging of Mitochondria in Living Cells: General Considerations
  • 5.Concluding Remarks: Other Potential Applications of Mito-PAGFP
  • Acknowledgments
  • References
  • 5.Characterization of Mitochondrial Transport in Neurons / Zu-Hang Sheng
  • 1.Introduction
  • 2.Analyses of Mitochondrial Transport in Embryonic Neuron Cultures
  • 3.Analyses of Mitochondrial Transport in Adult Neuron Cultures
  • 4.Characterization of Mitochondrial Transport at Synapses
  • 5.Summary
  • Acknowledgments
  • References
  • 6.Imaging of Mitochondrial Dynamics in Motor and Sensory Axons of Living Mice / Jordi Magrane
  • 1.Introduction
  • 2.Protocols to Image Mitochondrial Transport in the Peripheral Nerves of Living Mice
  • 3.Imaging of Mitochondrial Dynamics in the Peripheral Nerves of Living Mice
  • 4.Conclusions
  • Acknowledgments
  • References
  • 7.Analyzing Mitochondrial Dynamics in Mouse Organotypic Slice Cultures / David C. Chan
  • 1.Parasagittal Slice Cultures of the Basal Ganglia
  • 2.Cerebellar Slice Cultures
  • 3.Monitoring Mitochondrial Dynamics in Slice Cultures
  • References
  • 8.Analysis of Mitochondrial Traffic in Drosophila / Peter J. Hollenbeck
  • 1.Introduction
  • 2.Dissection Methods for Drosophila Larvae
  • 3.Imaging
  • 4.Analysis
  • 5.Conclusions
  • Acknowledgment
  • References
  • 9.In Vivo Imaging of Mitochondria in Intact Zebrafish Larvae / Thomas Misgeld
  • 1.Introduction
  • 2.Preparation of Transgenic MitoFish for Live Imaging
  • 3.Visualizing Axonal Transport of Mitochondria in Zebrafish Sensory Neurons
  • 4.Processing and Quantification of Imaging Files
  • 5.Conclusions
  • Acknowledgments
  • References
  • 10.The Use of miniSOG in the Localization of Mitochondrial Proteins / Guy A. Perkins
  • 1.Introduction
  • 2.Requirements for CLEM Labeling
  • 3.miniSOG Features
  • 4.Resolution
  • 5.Photooxidation Protocol for a Monolayer of Cultured Cells
  • 6.Photooxidation Protocol for Tissues
  • 7.Example of miniSOG Use with MCU
  • 8.Conclusions and Future Work
  • Acknowledgments
  • References
  • 11.Assessing the Function of Mitochondria-Associated ER Membranes / Estela Area-Gomez
  • 1.Introduction
  • 2.Isolation of MAM
  • 3.Assaying MAM Activity
  • References
  • 12.Measurement of ROS Homeostasis in Isolated Mitochondria / A. Ambrus
  • 1.Importance of Quantifying Mitochondrial ROS Homeostasis
  • 2.Detection of Mitochondrial ROS Formation
  • 3.Mitochondrial Elimination of H2O2
  • 4.Methods for the Detection of Oxidative Stress
  • Acknowledgments
  • References
  • 13.Use of Potentiometric Fluorophores in the Measurement of Mitochondrial Reactive Oxygen Species / Brian A. Roelofs
  • 1.Introduction
  • 2.ROS Detection Using MitoSOX
  • Subcellular Localization and Fluorescence Yield
  • 3.Validating MitoSOX Using a Negative Control
  • 4.Choosing a Correct MitoSOX Loading Paradigm
  • Additional Considerations
  • 5.Is MitoSOX Imaging Useful?
  • 6.Preparation of Primary Rat Cortical Neurons for Imaging
  • 7.Optimizing MitoSOX Concentration and Establishing Mitochondrial Localization
  • 8.Imaging Using MitoSOX
  • Acknowledgment
  • References
  • 14.Spatial, Temporal, and Quantitative Manipulation of Intracellular Hydrogen Peroxide in Cultured Cells / Rajiv R. Ratan
  • 1.Introduction
  • 2.Production of H2O2 Using DAAO
  • 3.Two-Photon Fluorescence Imaging of H2O2
  • 4.Summary
  • Acknowledgments
  • References
  • 15.Biochemical and Biophysical Methods for Studying Mitochondrial Iron Metabolism / Tracey A. Rouault
  • 1.Introduction
  • 2.Measurement of Total Iron Concentration
  • 3.In Situ Analysis of Iron in the Mitochondria
  • 4.Biophysical Methods for Studying Iron in Isolated Mitochondria
  • 5.Conclusions
  • Acknowledgments
  • References
  • 16.Analysis and Interpretation of Microplate-Based Oxygen Consumption and pH Data / Martin Jastroch
  • 1.Introduction
  • 2.The Oxygen Consumption Rate
  • 3.The Extracellular Acidification Rate
  • Summary
  • Acknowledgments
  • References
  • 17.Imaging Changes in the Cytosolic ATP-to-ADP Ratio / Gary Yellen
  • 1.Introduction
  • 2.Methods
  • 3.Expected Results
  • 4.Summary
  • References
  • 18.The Use of Mitochondria-Targeted Endonucleases to Manipulate mtDNA / Carlos T. Moraes
  • 1.Mitochondrial DNA
  • 2.Mitochondria-Targeted Restriction Nucleases to Cleave mtDNA and Model OXPHOS Diseases
  • 3.mtDNA Heteroplasmy and Approaches to Alter the Balance Between Wild-Type and Mutant mtDNA
  • 4.mtDNA Heteroplasmy Shift Using Restriction Endonucleases
  • 5.Designer Endonucleases for the Modulation of mtDNA Heteroplasmy
  • 6.mtDNA Heteroplasmy Shift Using Zinc-Finger Nucleases
  • 7.Heteroplasmy Shift Using TAL Effector Nucleases
  • 8.Single-Strand Annealing Assay to Analyze the Efficacy and Specificity of Designer Nuclease
  • 9.The Use of Cybrid Cells to Test Approaches to Change Mitochondrial DNA Heteroplasmy
  • 10.Immunodetection and Mitochondrial Localization in Cells and Tissues
  • 11.Changing mtDNA Heteroplasmy in Cultured Cells with mito-TALENs
  • 12.Evaluation of the mtDNA Content
  • 13.Future Perspectives
  • Acknowledgments
  • References
  • 19.Induced Pluripotent Stem Cell-Derived Models for mtDNA Diseases / Riikka H. Hamalainen
  • 1.Introduction
  • 2.Induced Pluripotent Stem Cells
  • 3.Mitochondrial Disease
  • 4.Conclusion
  • 5.Generation of iPSCs from mtDNA Disease Patients
  • Acknowledgment
  • References
  • 20.The Use of 18F-BCPP-EF as a PET Probe for Complex I Activity in the Brain / Hideo Tsukada
  • 1.Introduction
  • 2.Design and Assessment of Candidate Compounds for MC-I Imaging Probes
  • 3.Application for Stroke/Ischemic Damage Imaging
  • 4.Application for Imaging of Aging Effects on MC-I Activity
  • 5.Conclusion
  • References
  • 21.MR OEF Imaging in MELAS / Sheng Xie
  • 1.Introduction of Magnetic Resonance Imaging Techniques
  • 2.Significance of OEF Imaging in MELAS
  • 3.Cerebral OEF Changes In Our Study of MELAS
  • References.

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