Dr. Jaffe will continue to explore the roles of free cytosolic calcium pulses, waves and gradients in the development of embryological pattern, in cytokinesis as well as in fertilization. His broad purpose is to discover and describe spatiotemporal calcium patterns which have been conserved in evolution and to explore their developmental roles and mechanisms. His main method for visualizing these patterns will be to inject chemiluminescent, calcium specific proteins called aequorins. He will also use vibrating calcium specific electrodes to explore patterns of calcium currents through cell surfaces. He will continue this exploration in fucoid, ascidian, Xenopus and medaka eggs as well as in some other systems. One of the main methods for modifying, and thus for exploring the role of, these calcium patterns will be to suppress them by injecting appropriate calcium buffers. Dr. Jaffe will also modify such patterns via appropriate applications of calcium channel blockers and agonists as well as calcium ionophores. This enterprise should prove to be of broad value in understanding cell development because calcium ions are among a relatively small class of small substances (like hydrogen ions, ATP, cyclic AMP, IP3, etc.) whose intracellular patterns are central to the physiological control mechanisms within cells.