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Fig. 2. Ca2+ puff activity during local and global signalling. All of the cell types analysed in the present study displayed Ca2+ puffs when treated with appropriate agonist concentrations. The aim of this figure is to show some of the different modes in which Ca2+ puff activity can be observed in these cells. (A) Rapid Ca2+ puff activity in a 16HBE14o- cell stimulated with 5 µM ATP. Several Ca2+ puff sites were active before and during the onset of the Ca2+ wave. White circles on the inset cell image show the locations of the active puff sites, and the Ca2+ signals recorded at these regions are depicted by the correspondingly numbered traces. (B) A HeLa cell stimulated with 1 µM histamine and in which a single Ca2+ puff sites was responsible for triggering a regenerative Ca2+ wave. The black trace represents the Ca2+ signal observed at the puff site and the red trace depicts the global Ca2+ signal observed by averaging fluo-3 fluorescence across the whole cell. (C) Ca2+ puffs firing at three different sites within a single carbachol-stimulated SH-SY5Y cell (1 µM carbachol). Coloured circles on the inset cell image show the locations of the active puff sites, and the Ca2+ signals recorded at these regions are depicted by the correspondingly coloured traces. This example illustrates the observation that, sometimes, the Ca2+ puff sites appear to fire in synchrony, whereas, at other times, they do not. (D) Ca2+ puffs in a NIH-3T3 cell stimulated with 1 µM ATP. Ca2+ puffs were observed at the regions marked by the blue, green and red circles on the inset cell image, and the correspondingly coloured traces indicate the Ca2+ changes observed at these sites. Ca2+ puffs were observed during the initial latency before the first Ca2+ oscillation and also on the rising phases of subsequent Ca2+ oscillations.





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