Abstract
This chapter describes the use of tetracarboxylate- derived fluorescent Ca 2+ indicators to measure changes in the cytosolic free Ca 2+ in single cells and suspensions of cells. This is followed by a discussion of several approaches that can be used in conjunction with these indicators to study the biochemical mechanisms regulating the Ca 2+ signal. This chapter discusses the mechanisms account for the diversity of cellular Ca 2+ responses seen in different tissues, as well as the marked heterogeneity of responses seen in single cells within a population, including an oscillatory behavior of the Ca 2+ signal. An elucidation of the detailed molecular mechanisms underlying these regulatory processes requires that changes in the cytosolic Ca 2+ concentration be reliably measured. The receptor-mediated Ca 2+ signaling often involves both release of internal Ca 2+ stores and stimulation of Ca 2+ entry, experimental manipulations in which these two components are selectively inhibited need to be used.
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