The use of aequorins to record and visualize Ca2 Dynamics: From subcellular microdomains to whole organisms

Abstract

In this chapter, we describe the practical aspects of measuring [Ca2+] transients that are generated in a particular cytoplasmic domain, or within a specific organelle or its periorganellar environment, using bioluminescent, genetically encoded and targeted Ca2+ reporters, especially those based on apoaequorin. We also list examples of the organisms, tissues, and cells that have been transfected with apoaequorin or an apoaequorin-BRET complex, as well as of the organelles and subcellular domains that have been specifically targeted with these bioluminescent Ca2+ reporters. In addition, we summarize the various techniques used to load the apoaequorin cofactor, coelenterazine, and its analogs ..