In the first edition of Calcium Signaling Protocols I began by writing "The regula- 2+ tion of intracellular Ca is a common theme presented in many papers over the last 20 2+ or so years and the description of the Ca -sensitive indicator dye fura-2 in 1985 resulted in a massive increase in these types of studies. " This statement is as true in 2005 as it was in 1999, but 20 or so years is now 30 years! There has been some reorganization of the volume such that there are now 22 ch- ters including five new ones, all written by experts in their field. These new chapters 2+ include use of the FlexStation and electrophysiological measurement of Ca channel activity. The book is broken into six parts. Part I is a general coverage of basic theory and the simplest use of fluorescent indicators. Part II covers specialist measurement 2+ systems and Part III covers measurement of Ca channel activity. Assessment of 2+ release of stored Ca is covered in some detail in Part IV, with Parts V and VI cover- 2+ ing specialist measurement techniques and Ca -sensitive targets. Putting a book like this together, even as a second edition, takes time and I am, again, indebted to the individual authors for their help and patience. I am also very grateful to Professor John M. Walker, the series editor, for his continued help and advice over the course of this project.

Leseprobe

Part I. General Fluorescent Measurement of [Ca2+]c: Basic Practical Considerations Alec W. M. Simpson Measurement of [Ca2+]i in Whole Cell Suspensions Using Fura-2 Robert A. Hirst, Charlotte Harrison, Kazuyoshi Hirota, and David G. Lambert Measurement of [Ca2+]i in Cell Suspensions Using Indo-1 Adriaan Nelemans Part II. Specialist Measurement Systems Confocal Microscopy: Theory and Applications for Cellular Signaling Stephen C. Tovey, Paul J. Brighton, and Gary B. Willars Single-Cell and Subcellular Measurement of Intracellular Ca2+ Concentration Anthony J. Morgan and Andrew P. Thomas Ratiometric Ca2+ Measurements Using the FlexStation Scanning Fluorometer Ian C. B. Marshall, Izzy Boyfield, and Shaun McNulty Measuring Ca2+ Changes in Multiwell Format Using the Fluorometric Imaging Plate Reader Ian C. B. Marshall, Davina E. Owen, and Shaun McNulty Part III. Measurement of Ca2+ Channel Activity Measurement of Ca2+ Entry Using 45Ca2+ Mercedes Villarroya, Manuela G. Lopez, María F. Cano-Abad, and Antonio G. García Measurement of [3H]PN200-110 and [125I]w-Conotoxin MVIIA Binding Kazuyoshi Hirota and David G. Lambert Whole-Cell Patch Clamp Recording of Voltage-Sensitive Ca2+ Channel Currents: Heterologous Expression Systems and Dissociated Brain Neurons Atticus H. Hainsworth, Andrew D. Randall, and Alessandro Stefani Part IV. Measurement of Ins(1,4,5)P3 and Ca2+ Release From Intracellular Stores Measurement of Phospholipase C by Monitoring Inositol Phosphates Using [3H]Inositol-Labeling Protocols in Permeabilized Cells Alison Skippen, Philip Swigart, and Shamshad Cockcroft Measurement of Inositol(1,4,5)Triphosphate Using a Stereospecific Radioreceptor Mass Assay Darren Smart Measurement of Calcium Fluxes in Permeabilized Cells Using a 45Ca2+ Uptake and Release Assay Robert A. Wilcox Microinjection of myo-Inositol(1,4,5)Trisphosphate and Other Calcium-Mobilizing Agents Into Intact Adherent Cells Robert A. Wilcox, Ian D. Forsythe, and Terence J. McCann Measurement of Free [Ca2+] Changes in Agonist-Sensitive Internal Stores Using Compartmentalized Fluorescent Indicators Aldebaran M. Hofer Part V. Specialist Measurement Techniques Measurement of [Ca2+]i in Smooth Muscle Strips Using Front-Surface Fluorimetry Hideo Kanaide Measurement of Calcium and Movement in Heart Cells Leong L. Ng and Paulene A. Quinn Simultaneous Analysis of Intracellular pH and Ca2+ From Cell Populations Raul Martinez-Zaguilan, Linda S. Tompkins, Robert J. Gillies, and Ronald M. Lynch Measurement of Cytosolic-Free Ca2+ in Plant Tissue Martin R. McAinsh and Carl K.-Y. Ng Part VI. Ca2+-Sensitive Targets Assay and Purification of Calmodulin-Dependent Protein Kinase Rajendra K. Sharma Assay and Purification of Calmodulin-Dependent Cyclic Nucleotide Phosphodiesterase and Isozyme Separation Rajendra K. Sharma Measurement of Ca2+-ATPase Activity (in PMCA and SERCA1) Danuta Kosk-Kosicka Index