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Review
. 2022 Dec:108:102658.
doi: 10.1016/j.ceca.2022.102658. Epub 2022 Oct 11.

The evolution of organellar calcium mapping technologies

Matthew Zajac  1 , Souvik Modi  2 , Yamuna Krishnan  3
Affiliations
Review

The evolution of organellar calcium mapping technologies

Matthew Zajac et al. Cell Calcium. 2022 Dec.

Abstract

Intracellular Ca2+ fluxes are dynamically controlled by the co-involvement of multiple organellar pools of stored Ca2+. Endolysosomes are emerging as physiologically critical, yet underexplored, sources and sinks of intracellular Ca2+. Delineating the role of organelles in Ca2+ signaling has relied on chemical fluorescent probes and electrophysiological strategies. However, the acidic endolysosomal environment presents unique issues, which preclude the use of traditional chemical reporter strategies to map lumenal Ca2+. Here, we broadly address the current state of knowledge about organellar Ca2+ pools. We then outline the application of traditional probes, and their sensing paradigms. We then discuss how a new generation of probes overcomes the limitations of traditional Ca2+probes, emphasizing their ability to offer critical insights into endolysosomal Ca2+, and its feedback with other organellar pools.

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Figures

Fig. 1.
Fig. 1.
Schematic of organellar Ca2+ levels. Ca2+ levels at resting state (left-hand side) and after stimulation (right-hand side) [–35]. Progressively increasing Ca2+ levels are represented by an increasing colour gradient.
Fig. 2.
Fig. 2.
Different approaches to map organellar Ca2+. (A) Organellar Ca2+ is mapped indirectly by placing Ca2+ probes in the cytoplasm or anchoring them on the organelle surface. (B) Direct measurement needs Ca2+ indicators localized inside the organelle lumen.
Fig. 3.
Fig. 3.
Chemical structures of the various Ca2+ indicators, shaded according to their excitation wavelength from UV (Violet/Blue) to near infra-red (Red/Brown).
Fig. 4
Fig. 4
of GECIs are (A) FRET based (left) and (B) Intensity-based, relying on circularly permutated EGFP (cpGFP). The CaM domain in both classes undergoes a conformational change and binds the M13 domain in order to bind Ca2+ and re-organizes the probe.
Fig. 5.
Fig. 5.
Schematic of Membrane Contact Site (MCS) between organelles. At MCSs the distance between two organellar membranes are <30 nm and they act as domains for lipid or Ca2+ transfer across organelles.
See this image and copyright information in PMC

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