Calcium dynamics: analyzing the Ca2+ regulatory network in intact cells

Calcium signaling is critical for all cells. As a free ion (Ca(2+)), calcium links many physiological stimuli to their intracellular effectors by interacting with binding proteins whose occupancy determines the cellular effect of stimulation. Because binding site occupancy depends on the history of Ca(2+) concentration ([Ca(2+)]), Ca(2+) dynamics are critical. Calcium dynamics depend on the functional interplay between Ca(2+) transport and buffering systems whose activities depend nonlinearly on [Ca(2+)]. Thus, understanding Ca(2+) dynamics requires detailed information about these Ca(2+) handling systems and their regulation in intact cells. However, effective methods for measuring and characterizing intracellular Ca(2+) handling have not been available until recently. Using concepts relating voltage-gated ion-channel activity to membrane potential dynamics, we developed such methods to analyze Ca(2+) fluxes in intact cells. Here we describe this approach and applications to understanding depolarization-induced Ca(2+) responses in sympathetic neurons.