The Use of Aequorins to Record and Visualize Ca Dynamics: From Subcellular Microdomains to Whole Organisms - Methods in Cell Biology - page 259

CHAPTER 10
The Use of Aequorins to Record and
Visualize Ca 2 þ Dynamics: From Subcellular
Microdomains to Whole Organisms
Sarah E. Webb, * Kelly L. Rogers, † Eric Karplus, ‡ and
Andrew L. Miller *
*Biochemistry and Cell Biology Section and State Key Laboratory of Molecular Neuroscience
Division of Life Science
HKUST, Clear Water Bay
Kowloon, Hong Kong, PR China
† The Walter and Eliza Hall Institute of Medical Research
Parkville, Australia
‡ Science Wares Inc.
Falmouth
Massachusetts, USA
Abstract
I. Introduction
II. Expression of Apoaequorin, GFP-Apoaequorin, and Other Apoaequorin-Based
Spectral Variants in Cells, Tissues, and Whole Organisms
A. Expression of Apoaequorin
B. Expression of GFP-Apoaequorin
C. Summary of Section II
III. Introducing Coelenterazines into Cells, Tissues and Embryos
IV. Techniques for Detecting Aequorin Luminescence
V. Conclusions
References
Abstract
In this chapter, we describe the practical aspects of measuring [Ca 2 þ ]
transients that are generated in a particular cytoplasmic domain, or within a
specific organelle or its periorganellar environment, using bioluminescent,