A Practical Guide to the Study of Calcium in Living Cells

393 Pages·1994·18.98 MB·English

by  Richard Nuccitelli (Eds.)

Methods in Cell Biology
VOLUME 40
A Practical Guide to the Study of Calcium in
Living Cells
Series Editors
Leslie Wilson
Department of Biological Sciences
University of California, Santa Barbara
Santa Barbara, California
Paul Matsudaira
Whitehead Institute for Biomedical Research and
Department of Biology
Massachusetts Institute of Technology
Cambridge, Massachusetts
Methods in Cell Biology
VOLUME 40
A Practical Guide to the Study of Calcium in
Living Cells
Edited by
Richard Nuccitelli
Section of Molecular and Cellular Biology
Division of Biological Sciences
University of California
Davis, California
ACADEMIC PRESS
A Division of Harcourt Brace G Company
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Cover photograph: From Chapter 10 by Diliberto ef al. For details see Color Plate 6.
This book is printed on acid-free paper. @
Copyright 0 1994 by ACADEMIC PRESS, INC.
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PRINTED IN THE UNITED STATES OF AMERICA
94 9 5 9 6 91 98 99 EB 9 8 7 6 5 4 3 2 I
CONTENTS
Contributors xi
Preface xiii
PART I Ca2+B uffers and [Ca2+]iP erturbation Techniques
1. A Practical Guide to the Preparation of Ca2+ Buffers
Donald M . Bers, Chris W. Patton, and Richard Nuccitelli
I. Introduction 4
11. Which Ca2+ Buffer Should You Use? 4
111. Basic Mathematical Relationships 8
IV. Temperature, Ionic Strength, and pH Corrections 12
V. Ca2+ Measurement and Calibration Solutions 14
VI. Preparing the Buffer Solution 18
VII. MaxChelator (MAXC v6.50): An Easy-to-Use Computer Program for
Ca" Buffer Preparation 21
References 28
2. Photorelease Techniques for Raising or Lowering
Intracellular CaZ+
Robert Zucker
I . Introduction 32
11. Nitr Compounds 33
111. DM-Nitrophen 37
IV. Diazo Compounds 40
V. Introduction into Cells 43
VI. Light Sources 44
VII. Calibration 46
VIII. Purity and Toxicity 49
IX. Biological Applications 50
X. Conclusions 58
References 59
3. Pharmacological Tools for Perturbing Intracellular
Calcium Storage
David Thomas and Michad R . Hanley
I . Introduction 65
11. SERCA Family of Intracellular Ca2+ Pumps 66
V
vi Contents
111. Acute Effects of Ca” Pump Inhibitors on Intact Cells 75
IV. Chronic Effects of Ca2+ Pump Inhibitors 80
V. Potential Applications of Intracellular Ca” Pump Inhibitors in
Calcium Cell Biology 82
References 84
PART I1 Microelectrode Techniques for Measuring [Ca*+]i and
Ca2+F luxes
4. How to Make and to Use Calcium-Specific Mini- and
Microelectrodes
Stiphane Baudet, Leis Hove-Madsen, and Donald M . Bers
I. Introduction 94
11. General Considerations 94
111. Minielectrodes 96
IV. Microelectrodes 102
V. Troubleshooting 111
References 112
5. The Vibrating Ca2+E lectrode: A New Technique for
Detecting Plasma Membrane Regions of Ca2+I nflux and Efflux
PeterJ. S. Smith, Richard H . Sanger, and Lionel F . J a f e
1. Introduction 115
11. Principles of Vibrating Electrodes 117
111. Construction of Ion-Selective Probes 122
IV. Calibration and Sensitivity 124
V. Data Acquisition 127
VI. Data Analysis and Examples 129
References 133
6. Application of Patch Clamp Methods to the Study of Calcium
Currents and Calcium Channels
Colin A . Leech and George G. Holz, IV
1. Introduction 135
11. Pipettes and Solutions 137
111. Patch Configurations 138
IV. Reconstitution of Purified Channel Proteins 142
V. Expression Cloning of Channel Subunits 144
VI. Pancreatic /3 Cell: A Model System for Analysis of Ca2+ Signaling 144
References 148
Contents vii
PART I11 Fluorescence Techniques for Imaging [Ca2+]j
7. Practical Aspects of Measuring [Ca”] with
Fluorescent Indicators
Josrph P. Y . Kao
I. Introduction 155
11. Fluorescent Ca” Indicators 156
111. Loading Indicators into Cells 161
IV. Manipulation of [Ca”] 167
V. Conversion of Indicator Fluorescence Signal into Values of [Ca”], 174
VI. Concluding Remarks 180
References 180
8. Rapid Simultaneous Estimation of Intracellular
Calcium and pH
Stephen J . Morris, Thomas B . Wiegmann, Larry W . Welling, arid
Bibir M . Clironwall
I . Introduction 184
11. Simultaneous lntracellular Calcium and pH Measurements 185
111. Correction Procedures and Calculations 196
IV. Choosing Calcium and pH Indicator Dyes 200
V. Possible (but untried) Dye Combinations 205
V1. Equipment 209
VII. Results 214
References 219
9. Improved Spatial Resolution in Ratio Images Using
Computational Confocal Techniques
Thomas J . Keating and R . J ohn Cork
I. Introduction 22 1
11. Computational Confocal Image Processing 222
111. Point Spread Function 223
IV. Deblurring and Inverse Filtering Algorithms 226
V. Ratios of Processed Images 230
VI. Hardware and Software Requirements 233
VII. Conclusions 235
Appendix I: Fourier Transforms 236
Appendix 11: Theoretical CTF Equations 239
References 240
10. Confocal Imaging of Ca2+i n Cells
Pamela A . Diliberto, Xue Feng Wang, and Brian Herman
1. Introduction 244
11. Ca2+I ndicator Dyes 244
...
Contents
V l l l
Ill. Instrumentation 248
IV. Calibration 25 1
V. Ratiometric vs. Nonratiometric Measurements 254
VI. Image Processing 257
VII. Applications 258
References 26 1
11. Simultaneous Near Ultraviolet and Visible Excitation
Confocal Microscopy of Calcium Transients in
Xenopus Oocytes
Steven cirard and David E . Clapham
I . Introduction 263
11. Confocal Microscopy 265
Ill . Calibration of Instrumentation and Fluorescence Signals 272
IV. Calcium Signaling in Oocytes 274
V. Mobile Buffers 280
V1. Conclusions 281
References 283
PART IV Use of Aequorin for [Ca2+]i Imaging
12. Inexpensive Techniques for Measuring [Ca2+]iC hanges Using
a Photomultiplier Tube
Kenneth R . Robinson, Thomas J . Keating, and R .J ohn Cork
I . Introduction 287
11. Photomultipliers 288
Ill. Specific System for Cellular Fluorescence Measurements 29 1
IV. Aequorin Measurements 299
V. Future Directions 301
VI. Summary 30 1
Appendix I: List of Suppliers 302
References 303
13. Imaging [Ca2+]iw ith Aequorin Using a Photon
Imaging Detector
Andrew L. Miller, Eric Karplus, and Lionel F.Ja#e
1. Introduction 306
11. Rationale for Choosing between Aequorin Luminescence and
Fluorescence Ratio Imaging of Ca2+-Sensitive Fluorophores 306
Ill. Types of Aequorin 313
IV. Sources, Handling, and Storage of Aequorin 314
V. Methods for the Introduction of Aequorins into Cells 31 5
Vl. Techniques for Detecting Aequorin Luminescence for the Purpose of
~maging[C a2+], 325
Contents ix
VII. Troubleshooting Experimental Difficulties with Aequorin 334
VIII. Conclusions 335
References 335
14. Targeting Recombinant Aequorin to Specific
Intracellular Organelles
Rosario Rirzuro, Murisa Brini, and Tullio Pozzan
I . Principles of Subcellular Targeting 340
11 . Construction of Chimeric Aequorin cDNAs 341
111. Expression of Recombinant Aequorin 343
IV. Reconstitution of Functional Aequorin 349
V. Biochemical Analysis of Aequorin Compartmentalization 350
VI. I n Vivo Measurements 350
VII. Conclusions 354
Appendix I: Protocols 354
Keferences 357
Index 359
Contents of Previous Volumes 365