Intracellular calcium waves accompany neutrophil polarization, formylmethionylleucylphenylalanine stimulation, and phagocytosis: a high speed microscopy study.
Using high sensitivity fluorescence imaging with shutter speeds approximately 600,000 times faster than those of video frames, we have characterized Ca2+ waves within cells in exquisite detail to reveal Ca2+ signaling routes. Polarized neutrophils exhibited a counterclockwise rotating ryanodine-sensitive juxtamembrane Ca2+ wave during temporal calcium spikes. During stimulation with fMLP, a chemotactic factor, two Ca2+ waves traveling in opposite directions around the perimeter of the cell emanated from sites of stimulation (the clockwise wave is verapamil sensitive). Phagocytosed targets exhibit counterclockwise Ca2+ waves traveling about their periphery originating from the plasma membrane. This study: 1) outlines the technology to observe Ca2+ signaling circuitry within small living cells; 2) shows that extracellular spatial information in the form of a chemotactic factor gradient is transduced into intracellular chemical patterns, which provides fresh insights in signaling; 3) suggests that a line of communication exits between the cell surface and phagosomes; and 4) suggests that spatiotemporal Ca2+ patterns contribute to drug actions.
Kindzelskii
Andrei L
AL
Department of Biological Sciences, Wayne State University, Detroit, MI 48202, USA.
Petty
Howard R
HR
D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000502:physiology*; D020013:Calcium Signaling / Q000276:immunology*; D002413:Cations, Divalent / Q000378:metabolism; D016764:Cell Polarity / Q000276:immunology*; D002634:Chemotaxis, Leukocyte / Q000276:immunology; D004912:Erythrocytes / Q000276:immunology; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007211:Indoles / Q000378:metabolism; D007424:Intracellular Fluid / Q000378:metabolism / Q000502:physiology*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D015591:Microspectrophotometry / Q000295:instrumentation / Q000379:methods; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D018375:Neutrophil Activation / Q000276:immunology*; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D010587:Phagocytosis / Q000276:immunology*; D010588:Phagosomes / Q000378:metabolism / Q000502:physiology; D012756:Sheep
D002413:Cations, Divalent; D005456:Fluorescent Dyes; D007211:Indoles; D009240:N-Formylmethionine Leucyl-Phenylalanine; C048960:indo-1; D002118:Calcium