Measuring Calcium and Calmodulin Inside and Outside Cells

by Petersen, Ole Holger

• ISBN13: 9783540675365
• ISBN10: 3540675361
• Edition: 1st
• Format: Paperback
• Copyright: 2001-05-01
• Publisher: Springer Verlag

Summary

The study of signal transduction mechanisms has become one of the most important branches of biomedical science, indispensable to understanding the normal actions of hormones, neurotransmitters and other extracellular signalling molecules as well as very many pathophysiological processes including inflammation and cancer. In this volume, the various techniques for measuring Ca2+ in the cytosol, in the various organelles and in the immediate surroundings of individual cells are described in practical details by experts, on the basis of their personal experience. Considering that the protein calmodulin is the main target for Ca2+ action in most cell types, a chapter on tracking calmodulin inside cells, using fluorescence technology, has also been included.

Table of Contents

Part A Cytosolic Ca2+ Concentration Measurements	1	(102)
Principles of Fluorescence Measurements - Dyes and Hardware Required	3	(44)
Emil C. Toescu
Alexej Verkhratsky
Simple Principles of Imaging - Image Files, Image Processing and Image Analysis	47	(18)
Emil C. Toescu
Aequorin Measurements of Cytosolic Ca2+	65	(26)
C. Jane Dixon
Anne K. Green
Preparing Cells for Microinjection	66	(4)
Preparation of Aequorin Stock Solution	70	(2)
Aequorin Microdialysis	72	(2)
Filling Micropipettes for Injection	74	(1)
Microinjection of Aequorin Into a Single Cell	75	(3)
Signal Detection From a Single Aequorin-Injected Cell	78	(4)
Data Presentation	82	(9)
Electrophysiological Measurements Using Ca2+-Sensitive Microelectrodes	91	(12)
Roger C. Thomas
Part B Measuring Ca2+ Inside Organelles	103	(146)
Measuring Ca2+ in the Nucleoplasm of Intact Cells	105	(26)
Marisa Brini
Paolo Pinton
Paulo Magalhaes
Luisa Filippin
Tullio Pozzan
Rosario Rizzuto
Measuring Ca2+ in Isolated Nuclei	131	(16)
Oleg Gerasimenko
Ca2+ Measurement with Luminescent Probes in the Endoplasmic Reticulum	147	(18)
Javier Alvarez
Mayte Montero
Ca2+ Measurement with Fluorescent Probes in the Endoplasmic Reticulum	165	(22)
Aldebaran M. Hofer
Ca2+ Measurements in Mitochondria	187	(26)
Paolo Pinton
Robert Drummond
Paulo Magalhaes
Marisa Brini
Anna Chiesa
Tullio Pozzan
Rosario Rizzuto
Ca2+ Measurements in Secretory Granules	213	(18)
Oleg Gerasimenko
Ca2+ Measurement in Endosomes of Intact Cells	231	(18)
Julia V. Gerasimenko
Part C Miscellaneous	249	(42)
Measuring Ca2+ Extrusion from Single Cells	251	(16)
Pavel V. Belan
Tracking Calmodulin Inside Cells	267	(24)
Madeleine Craske
Injection of Labelled Calmodulin into Cells	270	(5)
Monitoring Ca2+-Induced Intracellular Calmodulin Movements	275	(8)
Monitoring the Ca2+-Calmodulin Reaction in Different Subcellular Compartments	283	(8)
Subject Index	291