Disease relevance of FURA-2

• Gerbil cerebral cortical synaptosomes loaded with the fluorescent calcium probe FURA-2 were used to study depolarization-induced presynaptic cytosolic free calcium concentration, as an in vitro model of cerebral ischemia [1].
• To characterize the changes in free cytosolic calcium evoked by noxious heat stimuli (< or =51 degrees C, 10s), we performed microfluorometric measurements in acutely dissociated small dorsal root ganglion neurons (< or =32.5 microm) of adult rats using the dye FURA-2 [2].
• METHODS: We studied the effect of gambierol in human neuroblastoma cells by using bis-oxonol to measure membrane potential and FURA-2 to monitor intracellular calcium [3].
• Isolated rat parenchymal, endothelial, and Kupffer cells were cultured and changes in intracellular calcium measured in vitro after acute hypothermia (5-8 degrees C) by fluorescence imaging using FURA-2 [4].

High impact information on FURA-2

• Intracellular calcium ([Ca2+]i) was measured in FURA 2-loaded endothelial cells plated on fibronectin or vitronectin [5].
• These supernatants were added to FURA-2-loaded eosinophils that had been preincubated with thioperamide and/or the PAF antagonist WEB-2086 [6].
• Co-cross-linking of CD4 and Fc gamma R by an anti-CD4 mAb (MAX.16H5) and by excess of goat anti-mouse Ig induced a biphasic increase of the free cytosolic Ca(2+)-concentration ([Ca2+]i) in the human monocytoid cell line U937 as measured by FURA-2 fluorescence [7].
• Gap junction-mediated cell coupling in different connexin-transfected HeLa cells was standardized by measuring the spreading of microinjected Mn(2+) that led to local quenching of FURA-2 fluorescence [8].
• None of the phosphatase inhibitors changed calcium signals from FURA-2-loaded synaptosomes either before or after depolarization [9].

Chemical compound and disease context of FURA-2

• 3. The authors could not detect any effect of proline on glutamate-mediated [CA+2]i levels using FURA-2 in synaptosomes or neuroblastoma cells [10].

Biological context of FURA-2

• Here, we have measured IL-8-induced Ca2+ signals in single human neutrophil granulocytes using the Ca2+ indicator dye FURA-2 AM and we have investigated the signal transduction that leads to these Ca2+ signals with various pharmacological tools [11].
• The FURA-2 fluorescence kinetics were recorded at 340-380 nm [12].
• As determined by FURA-2 measurement of [Ca2+]I, the cytotoxicity of palmitate on cardiomyocytes did not appear to be mediated through acute increases in [Ca2+]l. In contrast, the unsaturated fatty acid, arachidonic acid increased [Ca2+]l. The calcium ionophore ionomycin significantly (P < 0.05) increased palmitate-induced cardiomyocyte cell death [13].
• QUIN 2/AM, BAPTA/AM, EGTA/AM, and FURA 2/AM, at 5 microM, decreased LDH release (at 6 hr) from 41% to 4%, 21%, 26%, and 33%, and decreased lipid peroxidation (at 1 hr) from 1.0 to 0.1, 0.4, 0.6, and 0.8 nmol MDA/mg protein, respectively, after TBHP exposure [14].

Anatomical context of FURA-2

• As keratinocyte maturation is regulated by calcium levels, we measured hyperosmotic-stimulus-induced changes of intracellular calcium ([Ca2+]i) by single-cell image analysis employing FURA-2/AM [15].
• Pericytes in suspension were loaded with 2 micromol/l of the Ca2+-sensitive dye FURA-2/AM [12].
• In single smooth muscle cells loaded with FURA-2, TRIM reduced the increase in fluorescence ratio produced by Tg but had no effect on that produced by high K [16].
• To investigate the underlying membrane mechanisms, current recordings together with simultaneous calcium measurements using FURA-2 were performed in neurons isolated from rat dorsal root ganglia [17].
• 1. This study investigates the mechanism of CGRP-induced relaxation in intramural coronary arteries by determining the effect of CGRP on cytosolic Ca(2+) concentration ([Ca(2+)](i)) using FURA-2 technique [18].

Associations of FURA-2 with other chemical compounds

• Basal intracellular calcium concentration (FURA 2 assay) was concentration-dependently increased (maximum effect +77%) 30 min, but not 24h after a 10 min glutamate (10-100 microM) treatment, while the net increase induced by electrical stimulation (10 Hz, 10s) was consistently reduced (maximum effect -64%) [19].
• WIN 55,212-2 was also shown to inhibit the NMDA-induced increase in intracellular calcium concentration ([Ca(2+)](i)) indicated by FURA-2 fluorescence imaging in the same cultured neurons [20].
• By using the dye FURA-2 AM, the variations in cytosolic Ca(2+) were studied in peripheral human T lymphocytes isolated from extracted blood from healthy donors [21].
• Control groups included 6 hours incubation in 4 degrees C Krebs Henseleit solution (n = 5); 6 hours incubation in 26 degrees C FURA-2/AM (n = 5); and 6 hours incubation in epinephrine 10(-7) mol/L (n = 5) [22].
• Extracellular acidification reduces the Mn(2+) entry, measured by the slope of the quenching of FURA 2 fluorescence at the isoemissive wavelength of 360 nm [23].

Gene context of FURA-2

• The intracellular distribution of NF aggregates, SOD1 and nNOS was examined by confocal microscopy and NMDA-induced alterations in intracellular calcium levels using either Oregon green fluorescence or FURA-2 photometric imaging [24].
• The SABP also induces an increase in intracellular Ca2+ in spermatozoa, as seen by FURA-2 AM studies [25].
• Using FURA-2 microspectrofluorimetry, this study examines the mobilisation of calcium in CHO cells which had been stably transfected with the long isoform of the human NK1 receptor [26].
• The effects of different isoforms of platelet-derived growth factor (PDGF) on cellular growth and on cytoplasmic Ca2+ levels in mesangial cells using a microfluorometric system and the calcium indicator FURA-2 were investigated [27].

Analytical, diagnostic and therapeutic context of FURA-2

• Single cell analysis of calcium fluxes was performed by video-microscopy and ratio-imaging, after cell staining with the fluorescent calcium chelator FURA-2 [28].
• To determine the cellular mechanisms by which the endothelium-dependent relaxation occurs, we used fura-2 fluorometry (F350 and F390; excitation wavelengths, 350 and 390 nm, respectively) and estimated the intracellular Ba2+ concentration in endothelial and vascular smooth muscle cells [29].
• Further, we summarize aspects of the signal transduction cascades possibly linked to different receptor types of the SCO; these studies included the use of calcium imaging (FURA-2 technique), ELISA technique, and immunocytochemistry [30].
• Isometric tension development and intracellular free calcium concentration ([Ca(2+)](i)) were measured simultaneously in isolated vessel segments using wire myography and the FURA-2 fluorescence technique [31].

References