This expert volume covers an interdisciplinary and rapidly growing area of biomedical research comprising genetic, biochemical, pathological, and clinical studies aimed at the diagnosis and therapy of human diseases which are either caused by or associated with mitochondrial dysfunction. It dedicates itself to showcasing the tremendous efforts and the progress that has been made over the last decades in developing techniques and protocols for probing, imaging, and manipulating mitochondrial functions. Mitochondrial Medicine: Volume I, Probing Mitochondrial Function focuses on methods being…mehr
This expert volume covers an interdisciplinary and rapidly growing area of biomedical research comprising genetic, biochemical, pathological, and clinical studies aimed at the diagnosis and therapy of human diseases which are either caused by or associated with mitochondrial dysfunction. It dedicates itself to showcasing the tremendous efforts and the progress that has been made over the last decades in developing techniques and protocols for probing, imaging, and manipulating mitochondrial functions. Mitochondrial Medicine: Volume I, Probing Mitochondrial Function focuses on methods being used for the assessment of mitochondrial function under physiological conditions as well as in healthy isolated mitochondria. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshootingand avoiding known pitfalls.
Comprehensive and practical, Mitochondrial Medicine provides an essential source of know-how and inspiration to all researchers who are fascinated by this tiny organelle that seems so clearly to control the life and death of a single cell and whole organisms alike.
Preparation of 'Functional' Mitochondria: A Challenging Business.- Isolation and Quality Control of Functional Mitochondria.- Isolation of Mitochondria-Associated ER Membranes (MAMs) and Glycosphingolipid Enriched Microdomains (GEMs) from Brain Tissues and Neuronal Cells.- Fluorescence Analysis of Single Mitochondria with Nanofluidic Channels.- Optical Microwell Arrays for Large Scale Studies of Single Mitochondria Metabolic Responses.- Deep Resequencing of Mitochondrial DNA.- Single Cell Analysis of Mitochondrial DNA.- A High-Throughput Next-Generation Sequencing Assay for the Mitochondrial Genome.- Rapid Mitochondrial DNA Isolation Method for Direct Sequencing.- Analysis of Mitochondrial DNA and RNA Integrity by a Real-Time qPCR Based Method.- Mitochondria-Targeted RNA Import.- Accurate Measurement of Circulating Mitochondrial DNA Content from Human Blood Samples Using Real Time Quantitative PCR.- mTRIP: An Imaging Tool to Investigate Mitochondrial DNA Dynamics in Physiology and Disease at the Single-Cell Resolution.- Simultaneous Quantification of Mitochondrial ATP and ROS Production.- Live-Cell Assessment of Mitochondrial Reactive Oxygen Species Using Dihydroethidine.- Detection and Differentiation Between Peroxynitrite and Hydroperoxides Using Mitochondria-Targeted Arylboronic Acid.- Time-Resolved Spectrometry of Mitochondrial NAD(P)H Fluorescence and Its Applications for Evaluating the Oxidative State in Living Cells.- Novel Methods for Measuring the Mitochondrial Membrane Potential.- High-Throughput Real-Time Analysis of Cell Oxygenation Using Intracellular Oxygen-Sensitive Probes.- In Vivo Assessment of Mitochondrial Oxygen Consumption.- Imaging Mitochondrial Hydrogen Peroxide in Living Cells.- Simultaneous High-Resolution Measurement of Mitochondrial Respirationand Hydrogen Peroxide Production.- Measurement of Mitochondrial NADH and FAD Autofluorescence in Live Cells.- Mitochondrial Coenzyme Q10 Determination Via Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry.- Assessing the Bioenergetic Profile of Human Pluripotent Stem Cells.- Integrative Methods for Studying Cardiac Energetics.- Computer-Based Prediction of Mitochondria-Targeting Peptides.- Prediction of Mitochondrial Protein Function by Comparative Physiology and Phylogenetic Profiling.- Assessment of Posttranslational Modification of Mitochondrial Proteins.- Assessment of Mitochondrial Protein Glutathionylation as Signaling for CO Pathway.- High-Resolution Melting Analysis for Identifying Sequence Variations in Nuclear Genes for Assembly Factors and Structural Subunits of Cytochrome C Oxidase.- Heterologous Inferential Analysis (HIA) as a Method to Understand the Role of Mitochondrial rRNA Mutations in Pathogenesis.- Analysis of Mitochondrial Dysfunction During Cell Death.- The Use of FLIM-FRET for the Detection of Mitochondria-Associated Protein Interactions.- Assessment of Mitochondrial Ca2+ Uptake.- Qualitative Characterization of the Rat Liver Mitochondrial Lipidome Using All Ion Fragmentation on an Exactive Benchtop Orbitrap MS.- Characterization of Mitochondrial Populations During Stem Cell Differentiation.- An Ex Vivo Model for Studying Mitochondrial Trafficking in Neurons.
Preparation of 'Functional' Mitochondria: A Challenging Business.- Isolation and Quality Control of Functional Mitochondria.- Isolation of Mitochondria-Associated ER Membranes (MAMs) and Glycosphingolipid Enriched Microdomains (GEMs) from Brain Tissues and Neuronal Cells.- Fluorescence Analysis of Single Mitochondria with Nanofluidic Channels.- Optical Microwell Arrays for Large Scale Studies of Single Mitochondria Metabolic Responses.- Deep Resequencing of Mitochondrial DNA.- Single Cell Analysis of Mitochondrial DNA.- A High-Throughput Next-Generation Sequencing Assay for the Mitochondrial Genome.- Rapid Mitochondrial DNA Isolation Method for Direct Sequencing.- Analysis of Mitochondrial DNA and RNA Integrity by a Real-Time qPCR Based Method.- Mitochondria-Targeted RNA Import.- Accurate Measurement of Circulating Mitochondrial DNA Content from Human Blood Samples Using Real Time Quantitative PCR.- mTRIP: An Imaging Tool to Investigate Mitochondrial DNA Dynamics in Physiology and Disease at the Single-Cell Resolution.- Simultaneous Quantification of Mitochondrial ATP and ROS Production.- Live-Cell Assessment of Mitochondrial Reactive Oxygen Species Using Dihydroethidine.- Detection and Differentiation Between Peroxynitrite and Hydroperoxides Using Mitochondria-Targeted Arylboronic Acid.- Time-Resolved Spectrometry of Mitochondrial NAD(P)H Fluorescence and Its Applications for Evaluating the Oxidative State in Living Cells.- Novel Methods for Measuring the Mitochondrial Membrane Potential.- High-Throughput Real-Time Analysis of Cell Oxygenation Using Intracellular Oxygen-Sensitive Probes.- In Vivo Assessment of Mitochondrial Oxygen Consumption.- Imaging Mitochondrial Hydrogen Peroxide in Living Cells.- Simultaneous High-Resolution Measurement of Mitochondrial Respirationand Hydrogen Peroxide Production.- Measurement of Mitochondrial NADH and FAD Autofluorescence in Live Cells.- Mitochondrial Coenzyme Q10 Determination Via Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry.- Assessing the Bioenergetic Profile of Human Pluripotent Stem Cells.- Integrative Methods for Studying Cardiac Energetics.- Computer-Based Prediction of Mitochondria-Targeting Peptides.- Prediction of Mitochondrial Protein Function by Comparative Physiology and Phylogenetic Profiling.- Assessment of Posttranslational Modification of Mitochondrial Proteins.- Assessment of Mitochondrial Protein Glutathionylation as Signaling for CO Pathway.- High-Resolution Melting Analysis for Identifying Sequence Variations in Nuclear Genes for Assembly Factors and Structural Subunits of Cytochrome C Oxidase.- Heterologous Inferential Analysis (HIA) as a Method to Understand the Role of Mitochondrial rRNA Mutations in Pathogenesis.- Analysis of Mitochondrial Dysfunction During Cell Death.- The Use of FLIM-FRET for the Detection of Mitochondria-Associated Protein Interactions.- Assessment of Mitochondrial Ca2+ Uptake.- Qualitative Characterization of the Rat Liver Mitochondrial Lipidome Using All Ion Fragmentation on an Exactive Benchtop Orbitrap MS.- Characterization of Mitochondrial Populations During Stem Cell Differentiation.- An Ex Vivo Model for Studying Mitochondrial Trafficking in Neurons.
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