Introduction of aequorin into zebrafish embryos for recording Ca(2+) signaling during the first 48 h of development.

Ca(2+) signals, whether transient pulses, propagating waves, or long-duration, steady gradients, are generally considered to play an important role in the pattern-forming events that occur during vertebrate development. One vertebrate that has long been a favorite of embryologists because of its ex utero development and the optical clarity of its embryos is the zebrafish, Danio rerio. Using the bioluminescent Ca(2+) reporter aequorin, distinct Ca(2+) signals have been reported for at least the first 48 h of zebrafish development, with signals becoming progressively more complex as the embryo develops. Here we provide a general introduction to aequorin and its use in monitoring Ca(2+) signals and discuss methods for introducing aequorin into zebrafish embryos.