Dynamic imaging in living cells: windows into local signaling.

Highly localized changes in intracellular calcium concentration [Ca2+]i play a critical role in regulating numerous cellular functions, ranging from muscle contraction to neurotransmitter and hormone secretion to gene transcription. Fluorescent Ca2+ indicators have been invaluable tools in elucidating the role of localized changes in [Ca2+]i in regulating ion channels and other key proteins in various signaling pathways. Other techniques used to investigate localized changes in [Ca2+]i include approaches based on fluorescence resonance energy transfer, and electrophysiological measurements of ionic flux through Ca2+-sensitive channels. This Perspective discusses research using fluorescent Ca2+ indicators to study excitation-contraction coupling in cardiac myocytes, presenting both key findings and limitations of this approach. Complementary approaches useful in studying localized changes in Ca2+ and other second messengers (such as cyclic adenosine monophosphate) are also discussed.