Techniques for measuring and manipulating free Ca2+ in the cytosol and organelles of neutrophils.
Ca(2+) signalling in neutrophils is important for triggering and coordinating the behaviour of neutrophils. Fluorescent probes for cytosolic free Ca(2+) concentration, e.g., fura2 and fluo3, have been widely used in neutrophils. These probes can be used to monitor Ca(2+) in the cytosol, the nucleus, near the plasma membrane and theoretically within Ca(2+) storage organelles. The longer wavelength indicators, e.g., fluo3 and calcium green, can be used confocally to monitor subcellular Ca(2+) changes in the cytosol of neutrophils and in the nucleus. Confocal techniques also permit "impossible views" imaging of Ca(2+) and newer scanning techniques promise very fast temporal resolution. Techniques using chlortetracycline (CTC) and DiOC(6)(3) are also described for monitoring the position of Ca(2+) storage sites in neutrophils and for manipulating their activity. Thus, in this review, a spectrum of new (and older) optical techniques are presented which are useful for measuring, monitoring and manipulating cytosolic free Ca(2+) concentration and Ca(2+) storage in neutrophils. With these techniques, it is hoped that more insight will be gained into both the mechanism of and the consequences of Ca(2+) signalling in neutrophils.
Hallett
M B
MB
Molecular Signalling Group, University Department of Surgery, University of Wales College of Medicine, Heath Park, Cardiff, UK. hallettmb@cardiff.ac.uk
Hodges
R
R
Cadman
M
M
Blanchfield
H
H
Dewitt
S
S
Pettit
E J
EJ
Laffafian
I
I
Davies
E V
EV
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D003600:Cytosol / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008845:Microinjections; D018613:Microscopy, Confocal; D015336:Molecular Probe Techniques; D009504:Neutrophils / Q000737:chemistry* / Q000378:metabolism; D015388:Organelles / Q000737:chemistry / Q000378:metabolism*
D002118:Calcium