Brasen, Jens Christian3; Olsen, Lars Folke3; Hallett, Maurice B.2
1 Department of Biochemistry and Molecular Biology, Faculty of Science, SDU2 School of Medicine, Cardiff University3 Department of Biochemistry and Molecular Biology, Faculty of Science, SDU
In order to establish whether non-mitochondrial oxidase activity in human neutrophils is tightly related to cytosolic Ca2+ concentration, we simultaneously measured Ca2+ oscillations induced by ATP and oxidant production in single adherent neutrophils using confocal microscopy. ATP induced fast damped Ca2+ spikes with a period of 15 s and slower irregular spikes with a period greater than 50 s. Spikes in Ca2+ occurred in the absence of Ca2+ influx, but the amplitude was damped by inhibition of Ca2+ influx. Using the oxidation of hydroethidine as a cytosolic marker of oxidant production, we show that the generation of reactive oxygen species by neutrophils adherent to glass was accelerated by ATP. The step-up in NADPH oxidase activity followed the first elevation of cytosolic Ca2+ but, despite subsequent spikes in Ca2+ concentration, no oscillations in oxidase activity could be detected. ATP induced spikes in Ca2+ in a very reproducible way and we propose that the Ca2+ signal is an on-switch for oxidase activity, but the activity is apparently not directly correlated with spiking activity in cytosolic Ca2+.
B B a - Molecular Cell Research, 2011, Vol 1813, Issue 8, p. 1446-1452