Abstract |
Localized subcellular changes in Ca(2+) serve as important cellular signaling elements, regulating processes as diverse as neuronal excitability and gene expression. Studies of cellular Ca(2+) signaling have been greatly facilitated by the availability of fluorescent Ca(2+) indicators. The respective merits of different indicators to monitor bulk changes in cellular Ca(2+) levels have been widely evaluated, but a comprehensive comparison for their use in detecting and analyzing local, subcellular Ca(2+) signals is lacking. Here, we evaluated several fluorescent Ca(2+) indicators in the context of local Ca(2+) signals (puffs) evoked by inositol 1,4,5-trisphosphate (IP3) in cultured human neuroblastoma SH-SY5Y cells, using high-speed video-microscopy. Altogether, nine synthetic Ca(2+) dyes ( Fluo-4, Fluo-8, Fluo-8 high affinity, Fluo-8 low affinity, Oregon Green BAPTA-1, Cal-520, Rhod-4, Asante Calcium Red, and X-Rhod-1) and three genetically-encoded Ca(2+)-indicators (GCaMP6-slow, -medium and -fast variants) were tested; criteria include the magnitude, kinetics, signal-to-noise ratio and detection efficiency of local Ca(2+) puffs. Among these, we conclude that Cal-520 is the optimal indicator for detecting and faithfully tracking local events; that Rhod-4 is the red-emitting indicator of choice; and that none of the GCaMP6 variants are well suited for imaging subcellular Ca(2+) signals.
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Authors | Jeffrey T Lock, Ian Parker, Ian F Smith |
Journal | Cell calcium
(Cell Calcium)
Vol. 58
Issue 6
Pg. 638-48
(Dec 2015)
ISSN: 1532-1991 [Electronic] Netherlands |
PMID | 26572560
(Publication Type: Comparative Study, Journal Article, Research Support, N.I.H., Extramural)
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Copyright | Copyright © 2015 Elsevier Ltd. All rights reserved. |
Chemical References |
- Fluorescent Dyes
- Inositol 1,4,5-Trisphosphate Receptors
- Inositol 1,4,5-Trisphosphate
- Calcium
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Topics |
- Calcium
(metabolism)
- Calcium Signaling
(physiology)
- Cells, Cultured
- Fluorescent Dyes
(analysis)
- Humans
- Inositol 1,4,5-Trisphosphate
(metabolism)
- Inositol 1,4,5-Trisphosphate Receptors
(metabolism)
- Kinetics
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