Calcium signalling : a practical approach /

Calcium signalling : a practical approach /

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Bibliographic Details
Edition:2nd ed.
Imprint:Oxford ; New York : Oxford University Press, 2001.
Description:xix, 230 p., 10 p. of plates : ill. (some col.) ; 26 cm.
Language:English
Series:The practical approach series ;
Practical approach series.
Subject:
Calcium -- Physiological effect -- Research -- Methodology.
Calcium -- Analysis.
Second messengers (Biochemistry) -- Research -- Methodology.
Cellular signal transduction -- Research -- Methodology.
Calcium -- analysis.
Calcium Signaling.
Calcium Channels.
Fluorescent Dyes.
Photometry -- methods.
Calcium -- Analysis.
Cellular signal transduction -- Research -- Methodology.
Second messengers (Biochemistry) -- Research -- Methodology.
Format: Print Book
URL for this record:http://pi.lib.uchicago.edu/1001/cat/bib/4438694
Hidden Bibliographic Details
Other uniform titles:Cellular calcium.
Other authors / contributors:Tepikin, Alexei.
ISBN:0199638489 (hbk. : alk. paper)
0199638470 (pbk.)
Notes:Rev. ed. of: Cellular calcium / edited by James G. McCormack and P.H. Cobbold. c1991.
Includes bibliographical references and index.
Table of Contents:
  • Preface
  • List of protocols
  • Abbreviations
  • Part 1. New probes, new instruments, new methods
  • 1. Cameleons as cytosolic and intra-organellar calcium probes
  • 1. Introduction
  • 2. Cameleons as new generation indicators for Ca[superscript 2+]
  • Advantages of cameleons over synthetic fluorescent calcium chelators and photoprotein aequorin
  • Evolutions of cameleons
  • 3. Measurements of [Ca superscript 2+ subscript c] (cytosolic Ca[superscript 2+] concentration) using improved yellow cameleons
  • Improved yellow cameleons (yellow cameleons-2.1 and -3.1)
  • Concentration of yellow cameleons inside cells
  • Constitutive expression of cameleons and interaction with CaM and CaM-dependent enzymes
  • Photochromism of yellow fluorescent protein
  • 4. Measurements of nuclear Ca[superscript 2+] concentration ([Ca superscript 2+ subscript n])
  • [Ca superscript 2+ subscript c] versus [Ca superscript 2+ subscript n]
  • Targeting yellow cameleon-3.1 to nucleus
  • Sectional images of [Ca superscript 2+ subscript c] and [Ca superscript 2+ subscript n] by two-photon excitation
  • 5. Measurements of free Ca[superscript 2+] concentration inside the endoplasmic reticulum ([Ca superscript 2+ subscript er])
  • References
  • 2. Photometry, video imaging, confocal and multi-photon microscopy approaches in calcium signalling studies
  • 1. General introduction
  • 2. Fluorescent Ca[superscript 2+] indicators
  • Loading Ca[superscript 2+] indicators
  • Choice of Ca[superscript 2+] indicator
  • Using adherent cells
  • 3. Photometry and video imaging of Ca[superscript 2+]
  • Photomultiplier tubes
  • Cameras
  • Data format--counts and images
  • Intensity depth
  • Look-up tables
  • Photometry versus video imaging
  • Brightness and gain settings
  • 4. Troubleshooting photometry and video imaging
  • Image too dark
  • Image too bright
  • Indicator bleaching
  • 5. Single-photon laser scanning confocal microscopy
  • General introduction
  • General arrangement of a laser scanning confocal microscope
  • Spinning disc confocal microscopes
  • 6. Advantages and pitfalls of confocal microscopy
  • Speed considerations
  • Problems with changing cell shape
  • Intensity depth, gain, and off-set
  • 7. Examples
  • Linear Ca[superscript 2+] waves in image mode
  • Linear Ca[superscript 2+] wave in line scan mode
  • Elementary Ca[superscript 2+] signals
  • 8. Multi-photon confocal microscopy
  • Background
  • Hardware
  • 9. Multi-photon photolysis
  • 10. Data processing
  • Calibration of the indicator fluorescence
  • Self-ratio of single-wavelength indicators
  • Pseudocolour
  • Archiving
  • References
  • 3. Detecting and minimizing errors in calcium-probe measurements arising from transition metals and zinc
  • 1. Introduction
  • 2. Competition between endogenous transition metals and Ca[superscript 2+] for fluorescent probes
  • 3. Membrane-permeant transition metal chelators
  • 4. Determining if fluorometric measurements of Ca[superscript 2+] have been perturbed by transition metal ions
  • Acknowledgments
  • References
  • 4. Targeting of bioluminescent probes and calcium measurements in different subcellular compartments
  • 1. Introduction
  • 2. Background principles
  • 3. Designing chimeric aequorin variants
  • Bioluminescent probes for mitochondrial matrix and mitochondrial intermembrane space
  • Measurement of ER and SR calcium using targeted aequorin
  • Targeted aequorin as an instrument of calcium measurement in the Golgi apparatus
  • Cytoplasmic and plasma membrane targeted bioluminescent probes
  • Nuclear targeting of aequorin
  • 4. Use of aequorin in the presence of high Ca[superscript 2+] concentrations
  • 5. Concluding practical considerations
  • References
  • Part 2. Calcium measurement in different organelles
  • 5. Monitoring mitochondrial function in single cells
  • 1. Introduction
  • Historical perspective: why is mitochondrial function interesting in the context of a book on calcium signalling?
  • 2. Fundamentals
  • Mitochondrial metabolism
  • The relationship between [Delta gamma subscript m] and redox state
  • Consequences of mitochondrial depolarization: mitochondria as ATP consumers
  • Fluorescence measurement and imaging of mitochondrial function
  • 3. NADH and flavoprotein autofluorescence
  • Theoretical basis
  • Protocols
  • Validation and limitations
  • 4. Fluorimetric measurement of mitochondrial potential
  • Theoretical basis
  • Approaches to the use of indicators and technical requirements
  • Calibration
  • Validation
  • Toxicity
  • An interesting question?
  • 5. Measuring mitochondrial calcium ([Ca superscript 2+ subscript mt])
  • Introduction and theoretical basis
  • Protocols
  • Validation and limitations
  • 6. Measuring changes in [ATP]
  • Luciferase
  • [Mg superscript 2+ subscript i]
  • 7. Cytochrome absorptance measurements
  • Theoretical basis
  • Methods and limitations
  • Perspectives
  • 8. Simultaneous measurements of mitochondrial and other cellular parameters
  • Simultaneous monitoring of [Delta psi subscript m] and [Ca superscript 2+ subscript I]
  • Simultaneous recording of [Delta Psi subscript m] and autofluorescence
  • Simultaneous recording of [Ca superscript 2+ subscript cyt] and [Ca superscript 2+ subscript mt]
  • References
  • Appendix
  • 1. The mitochondriac's basic pharmacopoeia
  • 2. Inhibitors of mitochondrial respiration
  • Complex I
  • Complex III
  • Complex IV
  • Uncouplers
  • The F[subscript 1]F[subscript 0]-ATP(synth)ase
  • Electron donors
  • The adenine nucleotide translocase
  • The permeability transition pore
  • The calcium uniporter
  • The Na[superscript +]/Ca[superscript 2+] exchanger
  • 6. Using low-affinity fluorescent calcium indicators and chelators for monitoring and manipulating free [Ca superscript 2+] in the endoplasmic reticulum
  • 1. Introduction
  • Basic principles
  • Mechanism of dye uptake
  • Choice of indicator
  • 2. Experimental protocols (permeabilized cells)
  • Notes on calcium buffers
  • Experiments in patch clamped cells
  • Intact cell measurements
  • Calibration procedures
  • 3. Pitfalls
  • Problems of multiple compartments
  • Interference by Mg[superscript 2+] and other ions
  • 4. Advantages
  • 5. Manipulating intrastore [Ca superscript 2+] with a membrane-permeant Ca[superscript 2+] chelator, TPEN
  • Practical considerations for using TPEN
  • Applications
  • Acknowledgements
  • References
  • 7. Measuring calcium in the nuclear envelope and nucleoplasm
  • 1. Introduction
  • Techniques and experiments described in the chapter
  • Calcium transport in the nucleus
  • 2. Experimenting with isolated nuclei
  • Preparation of isolated mouse liver nuclei
  • Fluorescent labelling of nucleoplasm and nuclear envelope of isolated nuclei: measuring calcium signals in the nuclear envelope and nucleoplasm
  • 3. Measurements of nuclear and cytosolic Ca[superscript 2+] signals in intact isolated cells
  • References
  • Part 3. Monitoring specific calcium reactions
  • 8. Controlling cytoplasmic calcium and measuring calcium-dependent gene expression in intact cells
  • 1. Introduction
  • 2. Measuring gene expression in single cells
  • Reporter genes
  • 3. Generating defined cytoplasmic [Ca superscript 2+ subscript i] signals
  • Constructing a calcium clamp
  • Using calcium clamp
  • Characterizing and troubleshooting the Ca[superscript 2+] clamp
  • Summary and future perspectives
  • References
  • 9. Monitoring the generation and propagation of calcium signals in multicellular preparations
  • 1. Introduction
  • 2. Perfusion apparatus and imaging systems
  • Perfusion system
  • Membrane lung
  • Mixing chamber/bubble trap
  • Imaging chamber
  • Imaging devices
  • 3. Perfusion
  • Perfusion media
  • Tissue isolation
  • 4. Loading Ca[superscript 2+] indicator dyes
  • Loading protocol
  • Dye retention
  • Photobleaching
  • 5. Calibration
  • Calibration protocol
  • 6. Image acquisition and analysis
  • Image quality
  • Deconvolving non-confocal fura-2 images
  • 7. Future directions
  • Acknowledgments
  • References
  • 10. Pharmacological studies of new calcium release mechanisms
  • 1. Introduction
  • 2. Preparation of sea urchin egg homogenates and microsomes
  • Collection of sea urchin eggs
  • Sea urchin egg homogenates
  • Fractionation of sea urchin egg homogenate by Percoll density gradient
  • 3. Studying activation of calcium release channels
  • Fluorimetry of Ca[superscript 2+] release from egg homogenates
  • Testing microsomal fractions for Ca[superscript 2+] releasing properties
  • Pharmacology of cADPR signalling
  • Studying NAADP-induced Ca[superscript 2+] release
  • Desensitization of NAADP-induced calcium release
  • 4. Measuring production and degradation of second messengers that trigger calcium signals
  • Enzymatic synthesis and degradation of cADPR
  • NAADP metabolism
  • Detection of cADPR and NAADP metabolism by discontinuous bioassay
  • NGD assay: continuous assay for GDP-ribosyl cyclase activity
  • HPLC
  • Measurement of cADPR levels in cells and tissues
  • 5. Binding of agonists to calcium release channels
  • cADPR binding studies
  • The NAADP receptor
  • References
  • 11. Measuring calcium extrusion
  • 1. Introduction
  • 2. The droplet technique
  • Instruments and materials for the droplet technique
  • Droplet formation
  • Injection of substances into the droplet
  • Measurements of fluorescence changes of intracellular and extracellular indicators and calibration of calcium concentrations in the droplet experiment
  • Measuring droplet volume and volume of cells: calculation of calcium flux, rate of intracellular calcium concentration changes, and calcium binding capacity of the cytoplasm of the cell
  • 3. Calcium-sensitive jam technique--using calcium indicators bound to dextrans to measure calcium extrusion
  • Instruments and materials for calcium-sensitive jam technique
  • Calcium-sensitive gel
  • References
  • A1. Fluorescent calcium indicators and caged calcium probes discussed in this volume
  • A2. List of suppliers
  • Index
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