Monitoring of free calcium in the neuronal endoplasmic reticulum: an overview of modern approaches.

The concentration of free calcium within the lumen of the endoplasmic reticulum ([Ca2+]L) fluctuates between 100 and 1000 microM. High [Ca2+]L provides an electro-driving force for Ca2+ release and supports high Ca2+ diffusion rate within the endoplasmic reticulum lumen. Fluctuations in [Ca2+]L also regulate numerous chaperones, responsible for postranslational protein processing. Thus, [Ca2+]L integrates various signalling events and establishes a link between fast signalling, associated with the endoplasmic reticulum Ca2+release/uptake, and long-lasting adaptive responses relying primarily on the regulation of protein synthesis. This paper overviews modern approaches for the direct monitoring of [Ca2+]L which rely on three classes of low-affinity Ca2+ probes: ER-targeted aequorin, synthetic fluorescent Ca2+ dyes and GFP-based ER-targeted Ca2+ probes. These techniques, especially as applied to neurones, may substantially widen our appreciation of the endoplasmic reticulum as a universal signalling organelle.