Reference : A double-labelling fluorescent assay for concomitant measurements of [Ca2+]i and O2. ...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/10993/5730
A double-labelling fluorescent assay for concomitant measurements of [Ca2+]i and O2. production in human macrophages
English
Bueb, Jean-Luc mailto [University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit >]
Gallois, A. [> >]
Schneider, J. C. [> >]
Parini, J. P. [> >]
Tschirhart, Eric mailto [University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit >]
1995
Biochimica et Biophysica Acta
1244
1
79-84
Yes (verified by ORBilu)
0006-3002
[en] pharmacology ; Cell Differentiation ; Cell Line ; Dimethyl Sulfoxide ; Fura-2 ; Humans ; Macrophages ; Macrophages, Alveolar ; N-Formylmethionine Leucyl-Phenylalanine ; Rhodamine 123 ; Rhodamines ; Spectrometry, Fluorescence ; Superoxides ; metabolism ; drug effects ; diagnostic use ; Calcium
[en] To measure intracellular free Ca2+ concentration ([Ca2+]i) and superoxide (O2) production in human alveolar macrophages, we used the fluorescent Ca2+ indicator fura-2 and the O2-sensitive dye dihydrorhodamine-123, which becomes fluorescent in its oxidized form, rhodamine-123. We describe a new double-dye technique whereby the kinetics of both [Ca2+]i levels and O2. production can be monitored simultaneously. This technique was developed in the dimethylsulfoxide-differentiated monocytic-like U-937 cell line (not equal to U-937), validated by comparison with single dye measurements and applied to human alveolar macrophages. The chemotactic peptide N-formyl-L-Methionyl-L-Leucyl-L-Phenylalanine induced in both cell types a similar transient elevation in [Ca2+]i, followed within seconds by a sustained increase in O2 production, which was however 4-fold weaker in not equal to U-937 cells. These results indicate that O2 production is an early event following the stimulation of human alveolar macrophages. This new double-dye technique may be relevant to other O2 ion-producing cells and could help to define more precisely the kinetics of the events leading to this biological response.
http://hdl.handle.net/10993/5730

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