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Exp Physiol. 2010 Nov;95(11):1049-60. doi: 10.1113/expphysiol.2010.052456. Epub 2010 Aug 9.

Seeing is believing! Imaging Ca2+-signalling events in living cells.

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  • 1Centre for Vision and Vascular Sciences, Queen's University of Belfast, Institute of Clinical Sciences, Grosvenor Road, Royal Victoria Hospital, Belfast BT12 6BA, UK.


Ever since it was shown that maintenance of muscle contraction required the presence of extracellular Ca(2+), evidence has accumulated that Ca(2+) plays a crucial role in excitation-contraction coupling. This culminated in the use of the photoprotein aequorin to demonstrate that [Ca(2+)](i) increased after depolarization but before contraction in barnacle muscle. Green fluorescent protein was extracted from the same jellyfish as aequorin, so this work also has important historical links to the use of fluorescent proteins as markers in living cells. The subsequent development of cell-permeant Ca(2+) indicators resulted in a dramatic increase in related research, revealing Ca(2+) to be a ubiquitous cell signal. High-speed, confocal Ca(2+) imaging has now revealed subcellular detail not previously apparent, with the identification of Ca(2+) sparks. These act as building blocks for larger transients during excitation-contraction coupling in cardiac muscle, but their function in smooth muscle appears more diverse, with evidence suggesting both 'excitatory' and 'inhibitory' roles. Sparks can activate Ca(2+)-sensitive Cl() and K(+) currents, which exert positive and negative feedback, respectively, on global Ca(2+) signalling, through changes in membrane potential and activation of voltage-operated Ca(2+) channels. Calcium imaging has also demonstrated that agonists that appear to evoke relatively tonic increases in average [Ca(2+)](i) at the whole tissue level often stimulate much higher frequency phasic Ca(2+) oscillations at the cellular level. These findings may require re-evaluation of some of our models of Ca(2+) signalling to account for newly revealed cellular and subcellular detail. Future research in the field is likely to make increasing use of genetically coded Ca(2+) indicators expressed in an organelle- or tissue-specific manner.

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