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Ca2+ imaging of intracellular organelles: endoplasmic reticulum

Blum, Robert, Petersen, Ole Holger and Verkhratsky, Alexei 2010. Ca2+ imaging of intracellular organelles: endoplasmic reticulum. In: Verkhratsky, Alexei and Petersen, Ole Holger eds. Calcium measurement methods, Neuromethods, vol. 43. New York: Humana Press, pp. 146-167. (10.1007/978-1-60761-476-0_8)

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Abstract

The endoplasmic reticulum (ER) is a complex and highly dynamic three-dimensional intracellular membranous system, which acts as a dynamic calcium store in the majority of eukaryotic cells. The special arrangement of intra-ER Ca2+ buffers, characterized by low affinity for Ca2+, in combination with SERCA pump activity keeps intraluminal Ca2+ ([Ca2+]L) at ∼0.1–0.8 mM (Cell Calcium 38:303–310, 2005), thus creating a steep electrochemical gradient aimed at the cytosol. Activation of ER Ca2+ channels results in Ca2+ release, which contributes to [Ca2+]i elevation, whereas SERCA-dependent Ca2+ uptake assists termination of cytosolic Ca2+ signals. In addition, the continuous luminal space can act as a travelling route for free Ca2+ ions (“Ca2+ tunnels”), thus bypassing cytosolic Ca2+ buffers and preventing mitochondrial Ca2+ uptake or loss of Ca2+ over the plasma membrane. Furthermore, changes in [Ca2+]L regulate ER-resident chaperones, responsible for postranslational protein processing. Thus, [Ca2+]L integrates various signalling events and establishes a link between fast signalling, associated with the ER Ca2+release/uptake, and long-lasting adaptive responses relying primarily on the regulation of protein synthesis. This paper overviews modern techniques for the imaging of [Ca2+]L using synthetic fluorescent Ca2+ dyes. The methods for ER dye loading, with a particular emphasis on employment of ER targeted esterases (the Targeted-Esterase induced Dye loading, TED) to increase specific accumulation of the probes within the ER lumen are described in detail.

Item Type: Book Section
Date Type: Publication
Status: Published
Schools: Biosciences
Systems Immunity Research Institute (SIURI)
Subjects: Q Science > Q Science (General)
R Medicine > RC Internal medicine > RC0321 Neuroscience. Biological psychiatry. Neuropsychiatry
Uncontrolled Keywords: Calcium imaging ; Calcium indicator ; Esterase ; Endoplasmic reticulum ; Protein targeting ; Neurons
Additional Information: pbk ISBN: 9781617796777
Publisher: Humana Press
ISBN: 9781607614753
Related URLs:
Last Modified: 04 Jun 2017 03:13
URI: http://orca-mwe.cf.ac.uk/id/eprint/18284

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