Intracellular Ca2+ measurement with Indo-1 in substrate-attached cells: Advantages and special considerations

Wahl, Michael J.; Lucherini, M.J.; Gruenstein, Eric

doi:10.1016/0143-4160(90)90081-5

Cited by 86 publications

(50 citation statements)

References 14 publications

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“…Therefore, photobleaching is not a significant problem in these studies. This result is not surprising because previous workers have noted that photobleaching is not a significant problem for indo-1-labeled adherent cells for up to 30 min of observation (35).…”

Section: Figuresupporting

confidence: 48%

Intracellular Calcium Waves Accompany Neutrophil Polarization, Formylmethionylleucylphenylalanine Stimulation, and Phagocytosis: A High Speed Microscopy Study

Kindzelskii

Petty

2003

The Journal of Immunology

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Using high sensitivity fluorescence imaging with shutter speeds ∼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.

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Section: Figuresupporting

confidence: 48%

Intracellular Calcium Waves Accompany Neutrophil Polarization, Formylmethionylleucylphenylalanine Stimulation, and Phagocytosis: A High Speed Microscopy Study

Kindzelskii

Petty

2003

The Journal of Immunology

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“…Similarly the work of Zagari et al (1989) Grynkiewicz et al (1985), and the ratios of standards containing known amounts of Ca2' and fura-2 (free acid) were measured under the same conditions used in experiments on the microscope. In situ calibration using fura-2 gives similar fluorescence ratios, as has been shown in previous work (Williams et al, 1985;Wahl et al, 1990;Wahl et al, 1992). Indo-1.…”

Section: Introductionsupporting

confidence: 65%

“…Indo-1. Unlike the situation for fura-2, the in situ fluorescence ratio as a function of Ca2+ curve is significantly different from the curve obtained in vitro (Popov et al, 1988;Wahl et al, 1990). For (1985) and Williams et al (1985).…”

Section: Introductionmentioning

confidence: 55%

“…Although a number of studies have shown that stimulation of quiescent cells with growth factors causes an immediate rise in [Ca21]i (Rabinovitch et al, 1986;Vincentini and Villereal, 1986;Hesketh et al, 1988;Tucker et al, 1989;Wahl et al, 1990) This study is the first to show a direct relationship between early serum stimulated Cai2+ increase and subsequent DNA synthesis in human cells. It also goes beyond recent studies on BALB/3T3 cells by providing dose response data and demonstrating reversibility, which are strong indications of a cause and effect relationship.…”

mentioning

confidence: 88%

“…For (1985) and Williams et al (1985). Image analysis was performed as previously described (Johnson et al, 1990;Wahl et al, 1990 …”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Intracellular free Ca2+ in the cell cycle in human fibroblasts: transitions between G1 and G0 and progression into S phase.

Wahl

Gruenstein

1993

MBoC

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Intracellular free calcium ([Ca2+]i) has been proposed to play an important part in the regulation of the cell cycle. Although a number of studies have shown that stimulation of quiescent cells with growth factors causes an immediate rise in [Ca21]i (Rabinovitch et al., 1986;Vincentini and Villereal, 1986;Hesketh et al., 1988;Tucker et al., 1989;Wahl et al., 1990) This study is the first to show a direct relationship between early serum stimulated Cai2+ increase and subsequent DNA synthesis in human cells. It also goes beyond recent studies on BALB/3T3 cells by providing dose response data and demonstrating reversibility, which are strong indications of a cause and effect relationship.

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Separate analysis of nuclear and cytosolic Ca2+ concentrations in human umbilical vein endothelial cells

et al. 1996

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Ca2+ concentration inside human umbilical vein endothelial cells was studied separately in cytosol and nucleus by a confocal laser scanning microscopy using fluo-3. The in vivo calibration curve for cytosol and nucleus showed good linearity between fluorescence intensity and Ca2+ concentration in cytosol ([Ca2+]i) and nuclei ([Ca2+]n). After calibration, [Ca2+]n was constantly higher than [Ca2+]i before and after the chelation of extracellular Ca2+ suggesting an active Ca2+ accumulation system on nuclear membrane. [Ca2+]n was also constantly higher than [Ca2+]i after the stimulation of thrombin (0.05 U/ml), FCS (10%), and thapsigargin (Tsg, 1 microM). The temporal change of [Ca2+]n and [Ca2+]i was identical, and [Ca2+]i gradient towards the nucleus and peripheral or central [Ca2+]n rise was observed after these stimulations. From these results, [Ca2+]n is not only regulated by the active Ca2+ accumulation system on nuclear membrane at rest but also the generation of inositol-triphosphate. FCS caused heterogeneous [Ca2+]n or [Ca2+]i rise from cell to cell; single spike or oscillatory change of [Ca2+]n and [Ca2+]i was observed in about 56% of cells, which were completely abolished by the chelation of extracellular Ca2+, suggesting that FCS stimulated [Ca2+]n and [Ca2+]i rise solely depending on Ca2+ influx from extracellular medium. The higher concentration of [Ca2+]n and heterogeneous [Ca2+]n rise may have important roles in nuclear-specific cellular responses.

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Intracellular Ca2+ measurement with Indo-1 in substrate-attached cells: Advantages and special considerations

Cited by 86 publications

References 14 publications

Intracellular Calcium Waves Accompany Neutrophil Polarization, Formylmethionylleucylphenylalanine Stimulation, and Phagocytosis: A High Speed Microscopy Study

Intracellular Calcium Waves Accompany Neutrophil Polarization, Formylmethionylleucylphenylalanine Stimulation, and Phagocytosis: A High Speed Microscopy Study

Intracellular free Ca2+ in the cell cycle in human fibroblasts: transitions between G1 and G0 and progression into S phase.

Separate analysis of nuclear and cytosolic Ca2+ concentrations in human umbilical vein endothelial cells

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