Rescue of vasopressin V2 receptor mutants by chemical chaperones: specificity and mechanism.
AuthorsRobben JH,Sze M,Knoers NV,Deen PM
JournalMolecular biology of the cell
PubMed ID16267275
Because missense mutations in genetic diseases of membrane proteins often result in endoplasmic reticulum (ER) retention of functional proteins, drug-induced rescue of their cell surface expression and understanding the underlying mechanism are of clinical value. To study this, we tested chemical chaperones and sarco(endo)plasmic reticulum Ca(2+) ATPase pump inhibitors on ... More
Indo-1 binding to protein in permeabilized ventricular myocytes alters its spectral and Ca binding properties.
AuthorsHove-Madsen L, Bers DM
JournalBiophys J
PubMed ID1420876
We have examined the binding of the fluorescent Ca indicator indo-1 to cellular protein in permeabilized ventricular myocytes and also to soluble and particulate myocyte protein. Using either a filtration technique or equilibrium dialysis, and conditions similar to those in a cardiac myocyte patch clamped with 100 microM indo-1 in ... More
Long-term regulation of neuronal calcium currents by prolonged changes of membrane potential.
AuthorsFranklin JL, Fickbohm DJ, Willard AL
JournalJ Neurosci
PubMed ID1315850
'Although rapid-onset, short-term regulation of neuronal Ca currents by neurotransmitters and second messengers is well documented, little is known about conditions that can cause longer-lasting changes in Ca channel function. We report here that persistent depolarization is accompanied by slowly developing long-term reduction of neuronal Ca currents. Rat myenteric neurons ... More
Regulation of cytosolic free calcium concentration by intrasynaptic mitochondria.
AuthorsMartínez-Serrano A, Satrústegui J
JournalMol Biol Cell
PubMed ID1550964
'By the use of digitonin permeabilized presynaptic nerve terminals (synaptosomes), we have found that intrasynaptic mitochondria, when studied "in situ," i.e., surrounded by their cytosolic environment, are able to buffer calcium in a range of calcium concentrations close to those usually present in the cytosol of resting synaptosomes. Adenine nucleotides ... More
Intracellular astrocyte calcium waves in situ increase the frequency of spontaneous AMPA receptor currents in CA1 pyramidal neurons.
AuthorsFiacco TA, McCarthy KD
JournalJ Neurosci
PubMed ID14736858
'Spontaneous neurotransmitter release and activation of group I metabotropic glutamate receptors (mGluRs) each play a role in the plasticity of neuronal synapses. Astrocytes may contribute to short- and long-term synaptic changes by signaling to neurons via these processes. Spontaneous whole-cell AMPA receptor (AMPAR) currents were recorded in CA1 pyramidal cells ... More
L-type Ca2+ channel desensitization by F- reduces PhE-induced increase in [Ca2+]i but not stress.
AuthorsRatz PH, Lattanzio FA
JournalAm J Physiol
PubMed ID1372479
'F- (10 mM sodium fluoride plus deferoxamine to chelate contaminating aluminum) causes arterial contractions primarily by activating L-type Ca2+ channels. Results from the present study indicate that, although F(-)-induced contractions could be completely relaxed by washing out the F- with fresh buffer, a long-lasting effect of F- pretreatment was to ... More
Ca2+ oscillations induced by hormonal stimulation of individual fura-2-loaded hepatocytes.
AuthorsKawanishi T, Blank LM, Harootunian AT, Smith MT, Tsien RY
JournalJ Biol Chem
PubMed ID2473983
'Isolated rat hepatocytes were loaded with the Ca2+ indicator fura-2 to measure cytosolic free Ca2+ concentrations ([Ca2+]i) in individual cells by digital ratio imaging microscopy. Stimulation with 0.1 nM vasopressin, 0.5 microM phenylephrine, or 0.5 microM ATP caused repetitive spikes of high [Ca2+]i in a high percentage of cells, in ... More
Effects of activation on distribution of Ca2+ in single arterial smooth muscle cells. Determination with fura-2 digital imaging microscopy.
AuthorsGoldman WF, Wier WG, Blaustein MP
JournalCirc Res
PubMed ID2706759
'A rise in cytosolic free Ca2+ is the immediate trigger for contraction in mammalian vascular smooth muscle. We used the fluorescent calcium indicator fura-2 and digital imaging microscopy to study the spatial distribution of intracellular Ca2+ in arterial myocytes and the changes elicited by activation with norepinephrine (NE). Viable arterial ... More
Further characterization of the mechanisms mediating the rise in cytosolic free Na+ in thrombin-stimulated platelets. Evidence for inhibition of the Na+,K(+)-ATPase and for Na+ entry via a Ca2+ influx pathway.
AuthorsBorin M, Siffert W
JournalJ Biol Chem
PubMed ID1649180
'Human platelets were loaded with the fluorescent Na(+)-sensitive dye sodium-binding benzofuran isophtalate (SBFI), and changes in the fluorescence excited at 345 and 385 nm were analyzed after manipulations that evoked predictable changes in the cytosolic Na+ concentration ([Na+]i). Raising [Na+]i by either gramicidin D or monensin specifically increased the fluorescence ... More
Comparison of spectrum-shifting intracellular pH probes 5'(and 6')-carboxy-10-dimethylamino-3-hydroxyspiro[7H-benzo[c]xanthene-7, 1'(3'H)-isobenzofuran]-3'-one and 2',7'-biscarboxyethyl-5(and 6)-carboxyfluorescein.
AuthorsOwen CS
JournalAnal Biochem
PubMed ID1514696
'The dyes carboxy-SNARF-1 and BCECF are fluorescent probes of intracellular pH that exhibit changes in spectral shape upon proton binding which allow one to use measurements of fluorescence at two or more wavelengths in order to measure pH without artifacts associated with variability in dye loading, etc. In evaluating these ... More
Thapsigargin activates a calcium influx pathway in the unfertilized mouse egg and suppresses repetitive calcium transients in the fertilized egg.
AuthorsKline D, Kline JT
JournalJ Biol Chem
PubMed ID1387638
'At fertilization, the sperm initiates development of the mouse egg by inducing a large transient increase in the intracellular Ca2+ concentration ([Ca2+]i), which is followed by repetitive transient increases in [Ca2+]i. To determine how the repetitive Ca2+ transients are produced, thapsigargin, an inhibitor of the endoplasmic reticulum Ca-ATPase, was used ... More
Suppression by staurosporine of Ca(2+)-mobilization triggered by ligation of antigen-specific receptors on t and B lymphocytes. An essential role of protein tyrosine kinase in the signal transduction.
AuthorsYamashita Y, Hasegawa-Sasaki H, Sasaki T
JournalFEBS Lett
PubMed ID1879563
'It is known that the receptor for platelet-derived growth factor (PDGF) activates phospholipase C (PLC) by phosphorylating the gamma 1 isoform of PLC with the receptor protein-tyrosine kinase (PTK), whereas a guanine nucleotide-binding protein participates as a transducer in the PLC activation through the receptors for vasopressin, bombesin and prostaglandin ... More
Effects of auxin and abscisic acid on cytosolic calcium and pH in plant cells.
AuthorsGehring CA, Irving HR, Parish RW
JournalProc Natl Acad Sci U S A
PubMed ID11607135
'Dark-grown corn coleoptiles and parsley hypocotyls and their roots were loaded with acetoxymethyl esterified forms of the Ca2+ indicator fluo-3, and the pH indicator 2'',7''-bis (2-carboxyethyl)-5(and-6)-carboxyfluorescein. These tissues were treated with the plant growth regulator 2,4-dichlorophenoxyacetic acid (2,4-D), an auxin analogue, or abscisic acid (ABA), and the cytosolic pH (pHcyt) ... More
Optical detection of ion-channel-induced proton transport in supported phospholipid bilayers.
AuthorsYang TH, Yee CK, Amweg ML, Singh S, Kendall EL, Dattelbaum AM, Shreve AP, Brinker CJ, Parikh AN,
JournalNano Lett
PubMed ID17629349
'The integration of ion-channel transport functions with responses derived from nanostructured and nanoporous silica mesophase materials is demonstrated. Patterned thin-film mesophases consisting of alternating hydrophilic nanoporous regions and hydrophobic nanostructured regions allow for spatially localized proton transport via selective dimerization of gramicidin in lipid bilayers formed on the hydrophilic regions. ... More
Active involvement of Ca2+ in mitotic progression of Swiss 3T3 fibroblasts.
AuthorsKao JP, Alderton JM, Tsien RY, Steinhardt RA
JournalJ Cell Biol
PubMed ID2114410
'Global Ca2+ transients have been observed to precede nuclear envelope breakdown and the onset of anaphase in Swiss 3T3 fibroblasts in 8% (vol/vol) FBS. The occurrence of these Ca2+ transients was dependent on intracellular stores. These Ca2+ transients could be (a) abolished by serum removal without halting mitosis, and (b) ... More
Adenosine A1 receptor-induced inhibition of Ca2+ transients linked to action potentials in clonal pituitary cells.
AuthorsMollard P, Guérineau N, Chiavaroli C, Schlegel W, Cooper DM
JournalEur J Pharmacol
PubMed ID1680718
'Adenosine is a potent paracrine/autocrine feedback inhibitor of cell activation in a variety of tissues. Adenosine action was studied in pituitary cells, in which spontaneous electrical activity causes characteristic oscillations of the cytosolic free Ca2+ concentration, [Ca2+]i. Cells of the GH3B6 rat pituitary tumor line were studied by microspectrofluorimetry using ... More
Association of cytoplasmic free Ca2+ gradients with subcellular organelles.
AuthorsWahl M, Sleight RG, Gruenstein E
JournalJ Cell Physiol
PubMed ID1537888
'Previous investigations have identified gradients of intracellular free (Ca2+)i (Ca2+i) in the cytoplasm of human fibroblasts. In this study we have compared the spatial distribution of these gradients with the subcellular distribution of cytoplasmic organelles. Using the Ca(2+)-sensitive dye fura-2 and organelle-specific fluorescent dyes, we have found that the highest ... More
Spontaneous Ca2+ spiking in a vascular smooth muscle cell line is independent of the release of intracellular Ca2+ stores.
AuthorsByron KL, Taylor CW
JournalJ Biol Chem
PubMed ID8463226
'Monolayers of fura-2-loaded A7r5 cells, a cell line derived from rat embryonic aorta, generated spontaneous Ca2+ spikes that were synchronized within the cell population. These Ca2+ spikes were abolished by removal of extracellular Ca2+ or addition of nimodipine (50 nM), and their frequency was increased by depolarization with high K+ ... More
Stimulation by thrombin increases the cytosolic free Na+ concentration in human platelets. Studies with the novel fluorescent cytosolic Na+ indicator sodium-binding benzofuran isophthalate.
AuthorsBorin M, Siffert W
JournalJ Biol Chem
PubMed ID2174043
'The new fluorescent Na+ indicator sodium-binding benzofuran isophthalate (SBFI) was used for determination of the cytosolic free Na+ concentration, [Na+]i, in human platelets. The dye could be loaded into platelets in the form of its acetoxymethyl ester (SBFI-AM). Calibration of the fluorescence in terms of [Na+]i was done by measuring ... More
Fluorescence measurements of cytosolic free Na concentration, influx and efflux in gastric cells.
AuthorsNegulescu PA, Harootunian A, Tsien RY, Machen TE
JournalCell Regul
PubMed ID1712635
'Regulation of cytosolic free Na (Nai) was measured in isolated rabbit gastric glands with the use of a recently developed fluorescent indicator for sodium, SBFI. Intracellular loading of the indicator was achieved by incubation with an acetoxymethyl ester of the dye. Digital imaging of fluorescence was used to monitor Nai ... More
A confocal scanning laser microscope for quantitative ratiometric 3D measurements of [Ca2+] and Ca2+ diffusions in living cells stained with Fura-2.
AuthorsHelm PJ, Franksson O, Carlsson K
JournalPflugers Arch
PubMed ID7792144
'A confocal scanning laser microscope (CSLM) for observation and quantitative ratiometric measurements of the intracellular dynamics of Ca2+ ions in living neurons has been developed. The instrument consists of a UV-enhanced CSLM, an optical arrangement providing simultaneous excitation at two wavelengths, an electronic arrangement for processing the simultaneous fluorescence response, ... More
Bradykinin-induced Ca2+ entry, release, and refilling of intracellular Ca2+ stores. Relationships revealed by image analysis of individual human fibroblasts.
AuthorsByron KL, Babnigg G, Villereal ML
JournalJ Biol Chem
PubMed ID1730576
'Lys-Bradykinin (BK), a mitogen for human foreskin fibroblasts (HSWP cells) (Owen, N. E., and Villereal, M. L. (1983) Cell 32, 979-985), elicits a rapid, transient elevation of intracellular free Ca2+ concentration ([Ca2+]i) in these cells. We have used image analysis of fura-2-loaded HSWP cells to examine the BK-induced [Ca2+]i changes ... More
Dynamics of Ca2+ transients in norepinephrine-stimulated individual H-35 hepatoma cells: fura-2 digital imaging microscopy and high time-resolution microspectrofluorometry.
AuthorsYodozawa S, Tsunoda Y, Funai T, Tashiro Y
JournalJ Histochem Cytochem
PubMed ID1940304
'We investigated spatiotemporal changes in cytoplasmic free Ca2+ concentration ([Ca2+]i) in norepinephrine (NE)-stimulated and fura-2-loaded individual H-35 rat hepatoma cells, using digital imaging microscopy and high time-resolution microspectrofluorometry. Application of NE (5 x 10(-6) M) resulted in an initial transient increase in [Ca2+]i, followed by a small sustained [Ca2+]i plateau ... More
Pertussis toxin effects on chemoattractant-induced response heterogeneity in human PMNs utilizing Fluo-3 and flow cytometry.
AuthorsOmann GM, Harter JM
JournalCytometry
PubMed ID2036919
'Flow cytometric methods were utilized to determine N-formylpeptide-induced cytosolic calcium levels in human polymorphonuclear leukocytes (PMNs) detected with the calcium indicator Fluo-3. Fluo-3 was readily loaded into PMNs as the acetoxymethyl ester. At room temperature Fluo-3 extrusion was minimal (less than 10%) over a 2 h time period. Flow cytometric ... More
Changes in phospholipid composition and calcium flux in LLC-PK cells cultured at low magnesium concentrations.
AuthorsMahfouz MM, Smith TL, Kummerow FA
JournalBiochim Biophys Acta
PubMed ID2804073
'Monolayers of porcine kidney cells (LLC-PK) were grown in a series of Nu-Serum-supplemented media containing different Mg(2+) concentrations (480, 250, 25, 6.3 or 2.6 microM) to study the effect of Mg(2+) depletion on cellular phospholipid changes and the consequent effect on the membrane permeability to Ca(2+). Cells grown on 6.3 ... More
Total internal reflection fluorescence flow cytometry.
AuthorsWang J, Bao N, Paris LL, Geahlen RL, Lu C,
JournalAnal Chem
PubMed ID19007249
'Total internal reflection fluorescence microscopy (TIRFM) has been widely used to explore biological events that are close to the cell membrane by illuminating fluorescent molecules using the evanescent wave. However, TIRFM is typically limited to the examination of a low number of cells, and the results do not reveal potential ... More
Nicotinic receptors that bind alpha-bungarotoxin on neurons raise intracellular free Ca2+.
AuthorsVijayaraghavan S, Pugh PC, Zhang ZW, Rathouz MM, Berg DK
JournalNeuron
PubMed ID1310863
'Many populations of vertebrate neurons have a membrane component that binds alpha-bungarotoxin and cholinergic ligands. Despite the abundance of this component and its similarities to nicotinic receptors, its function has remained controversial. Using a fluorescence assay, we show here that activation of the component elevates the intracellular concentration of free ... More
Phase-dependent contributions from Ca2+ entry and Ca2+ release to caffeine-induced [Ca2+]i oscillations in bullfrog sympathetic neurons.
AuthorsFriel DD, Tsien RW
JournalNeuron
PubMed ID1610566
'Sympathetic neurons display robust [Ca2+]i oscillations in response to caffeine and mild depolarization. Oscillations occur at constant membrane potential, ruling out voltage-dependent changes in plasma membrane conductance. They are terminated by ryanodine, implicating Ca(2+)-induced Ca2+ release. Ca2+ entry is necessary for sustained oscillatory activity, but its importance varies within the ... More
Effects of tissue absorbance on NAD(P)H and Indo-1 fluorescence from perfused rabbit hearts.
AuthorsFralix TA, Heineman FW, Balaban RS
JournalFEBS Lett
PubMed ID2335209
'The effects of tissue optical absorbance on intracellular NAD(P)H and Indo-1 fluorescence emission have been evaluated in the perfused rabbit heart. These results demonstrate that the tissue optical absorbance significantly modifies the emission characteristics of these fluorophores. This tissue ''inner filter'' effect, observed with both probes, changed as a function ... More
The role of [Ca2+]i and [Ca2+] sensitization in the caffeine contracture of rat myocytes: measurement of [Ca2+]i and [caffeine]i.
AuthorsO'Neill SC, Donoso P, Eisner DA
JournalJ Physiol
PubMed ID2213589
'1. Fluorescence measurements have been made in single, isolated rat ventricular myocytes using the Ca2(+)-sensitive indicators Fura-2 and Indo-1. In Fura-2-loaded cells, the application of caffeine (2-20 mM) produced a change of fluorescence indicating an increase of [Ca2+]i which then spontaneously decayed to control levels. These changes of [Ca2+]i were ... More
'Cytosolic calcium ([Ca2+]i) is an important mediator of neuronal signal transduction, participating in diverse biochemical reactions that elicit changes in synaptic efficacy, metabolic rate, and gene transcription. Excessive [Ca2+]i also has been implicated as a cause of acute neuronal injury, although measurement of [Ca2+]i in living neurons by fluorescent calcium ... More
Improved fluorescent (calcium indicator) dye uptake in brain slices by blocking multidrug resistance transporters.
AuthorsManzini I, Schweer TS, Schild D,
JournalJ Neurosci Methods
PubMed ID17767961
'ATP-binding cassette (ABC) transporters are a family of transmembrane proteins that, also known as multidrug resistance proteins, transport a wide variety of substrates across biological membranes in an energy-dependent manner. Recently it has been shown that members of this protein family interfere with fluorescent (calcium indicator) dye uptake in taste ... More
Spectral evidence for non-calcium interactions of intracellular Indo-1.
AuthorsOwen CS, Shuler RL
JournalBiochem Biophys Res Commun
PubMed ID2775270
'Indo-1 is widely used to measure intracellular free calcium, [Ca2+]i, by comparing the fluorescence emission at 2 or more wavelengths with the emissions, which are assumed to be known, of Indo-1 when it is fully calcium-bound and when it is fully calcium-free. Accurate quantitation requires that these "reference" values be ... More
Intracellular diffusion, binding, and compartmentalization of the fluorescent calcium indicators indo-1 and fura-2.
AuthorsBlatter LA, Wier WG
JournalBiophys J
PubMed ID2275965
'We studied intracellular binding and possible compartmentalization of the fluorescent Ca2+ indicators, indo-1 and fura-2, in single mammalian cardiac ventricular cells that had been loaded with indo-1 and fura-2 by exposure to the acetoxymethylester form of the indicators (indo-1/AM and fura-2/AM). Techniques similar to those used in experiments on fluorescence ... More
Extracellular ATP as a trigger for apoptosis or programmed cell death.
AuthorsZheng LM, Zychlinsky A, Liu CC, Ojcius DM, Young JD
JournalJ Cell Biol
PubMed ID1988462
'Extracellular ATP is shown here to induce programmed cell death (or apoptosis) in thymocytes and certain tumor cell lines. EM studies indicate that the ATP-induced death of thymocytes and susceptible tumor cells follows morphological changes usually associated with glucocorticoid-induced apoptosis of thymocytes. These changes include condensation of chromatin, blebbing of ... More
Adsorption of pluronic F-127 on surfaces with different hydrophobicities probed by quartz crystal microbalance with dissipation.
AuthorsNejadnik MR, Olsson AL, Sharma PK, van der Mei HC, Norde W, Busscher HJ,
JournalLangmuir
PubMed ID19374344
'Triblock copolymers of polyethylene oxide (PEO) and polypropylene oxide (PPO), that is, PEOn-PPOm-PEOn, better known as Pluronic can adsorb to surfaces in either a pancake or a brushlike configuration. The brushlike configuration is advantageous in numerous applications, since it constitutes a surface repellent to proteins and microorganisms. The conformation of ... More
Multiple types of calcium channels in acutely isolated rat neostriatal neurons.
AuthorsHoehn K, Watson TW, MacVicar BA
JournalJ Neurosci
PubMed ID8382736
'Voltage-activated high- and low-threshold Ca2+ currents were studied using whole-cell voltage-clamp techniques and fura-2 fluorescence measurements of intracellular Ca2+ in neurons acutely isolated from rat neostriatum. High-threshold Ca2+ currents activated around -40 mV and were present in at least 95% of neostriatal neurons. The maximum current, 736 +/- 44 pA ... More
Thrombin-induced calcium oscillation in human platelets and MEG-01, a megakaryoblastic leukemia cell line.
AuthorsOzaki Y, Yatomi Y, Wakasugi S, Shirasawa Y, Saito H, Kume S
JournalBiochem Biophys Res Commun
PubMed ID1550592
'Digital imaging microscopy revealed that human platelets show periodic intracellular Ca++ elevation in response to 0.01 U/ml thrombin. MEG-01, a megakaryoblastic leukemia cell line, also responded with oscillatory intracellular Ca++ elevation (0.7-1 times/min) to thrombin (0.001-0.003U/ml). Ca++ transients appears to be fused with higher thrombin doses. With extracellular Ca++ concentrations ... More
Heat shock protein synthesis and cytoskeletal rearrangements occur independently of intracellular free calcium increases in Drosophila cells and tissues.
AuthorsDrummond IA, Livingstone D, Steinhardt RA
JournalRadiat Res
PubMed ID3126536
'Heat shock causes significant changes in intracellular free calcium ([Ca2+]i) which occur rapidly following temperature elevation. The resting level of free calcium in single Drosophila melanogaster larval salivary gland cells measured with the fluorescent indicator fura-2 is 198 +/- 31 nM (n = 4). It increases approximately 10-fold to 1870 ... More
Bradykinin stimulates Ca2+ mobilization in NCB-20 cells leading to direct inhibition of adenylylcyclase. A novel mechanism for inhibition of cAMP production.
AuthorsBoyajian CL, Garritsen A, Cooper DM
JournalJ Biol Chem
PubMed ID1848232
'The modulation of neuronal adenylylcyclase by Ca2+, acting via calmodulin, is a long-established example of a positive interaction between the Ca2(+)-mobilizing and cAMP-generating systems. In the present study, concentrations of Ca2+ that stimulate brain adenylylcyclase inhibit the adenylylcyclase of NCB-20 plasma membranes. These inhibitory effects of Ca2+ have been characterized ... More
Cytosolic free magnesium in cardiac myocytes: identification of a Mg2+ influx pathway.
AuthorsQuamme GA, Rabkin SW
JournalBiochem Biophys Res Commun
PubMed ID2322282
'Regulation of intracellular Mg2+ activity in the heart is not well characterized. Cardiac myocytes were prepared as primary cultures from 7 day old chick embryo hearts and intracellular Mg2+ concentration [( Mg2+]i) was determined in single ventricular cells with mag-fura-2. Basal [Mg2+]i was 0.48 +/- 0.03 mM in normal culture ... More
Galanin evokes a cytosolic calcium bursting mode and hormone release in GH3/B6 pituitary cells.
AuthorsGuérineau N, Drouhault R, Corcuff JB, Vacher AM, Vilayleck N, Mollard P
JournalFEBS Lett
PubMed ID1702391
'The effects of galanin on secretion and cytosolic free Ca2+ concentration ([Ca2+]i) have been studied in GH3/B6 pituitary cells. Prolactin (PRL) and growth hormone (GH) release was measured in column perifusion experiments; [Ca2+]i was monitored in single cells by dual emission microspectrofluorimetry using indo-1 as intracellular Ca2+ probe. Galanin (0.1-1 ... More
Dihydropyridine-sensitive L-type Ca2+ channels in human foreskin fibroblast cells. Characterization of activation with the growth factor Lys-bradykinin.
AuthorsBaumgarten LB, Toscas K, Villereal ML
JournalJ Biol Chem
PubMed ID1375223
'We have previously characterized the calcium response of cultured human fibroblasts (HSWP cells) to stimulation by the mitogen Lys-bradykinin (BK). We have reported a biphasic response which includes a rapid rise to a peak that appears to result from mobilization of internal calcium, and a plateau phase, which is due ... More
Inhibition of calcium mobilization is an early event in opiate-induced immunosuppression.
AuthorsSei Y, McIntyre T, Fride E, Yoshimoto K, Skolnick P, Arora PK
JournalFASEB J
PubMed ID2022315
'Morphine administered as a subcutaneous implant inhibits the initial increase in cytoplasmic free-calcium [Ca2+]i induced by mitogens in mouse splenocytes. This effect was not reproduced by incubation of splenocytes with morphine (10(-8)-10(-4) M). Analysis of splenocyte subpopulations demonstrates that this effect was manifest in both B and T cells. However, ... More
A delayed and sustained rise of cytosolic calcium is elicited by oxidized LDL in cultured bovine aortic endothelial cells.
AuthorsNègre-Salvayre A, Fitoussi G, Réaud V, Pieraggi MT, Thiers JC, Salvayre R
JournalFEBS Lett
PubMed ID1544476
'Bovine aortic endothelial cells (BAEC) pulsed for 5 h with mildly oxidized low density lipoproteins (LDL), exhibited a broad, sustained and high peak of [Ca2+]i occurring several hours after the end of the pulse and reaching very high [Ca2+]i values (around 2500-3000 nmol/l) and a concomitant drop of cytosolic pH ... More
Flow-induced calcium transients in single endothelial cells: spatial and temporal analysis.
AuthorsGeiger RV, Berk BC, Alexander RW, Nerem RM
JournalAm J Physiol
PubMed ID1616008
'Endothelial cells (EC) are uniquely situated to respond to hemodynamic forces. Because flow-mediated release of endothelial-derived relaxing factors is associated with increased EC intracellular calcium ([Ca2+]i), we sought to determine the effects of fluid shear stress on EC [Ca2+]i. Cells were subjected to flow in parallel-plate flow chambers and glass ... More
Imaging of cytosolic Ca2+ transients arising from Ca2+ stores and Ca2+ channels in sympathetic neurons.
AuthorsLipscombe D, Madison DV, Poenie M, Reuter H, Tsien RW, Tsien RY
JournalNeuron
PubMed ID2856095
'Changes in cytosolic free Ca2+ concentration [( Ca2+]i) due to Ca2+ entry or Ca2+ release from internal stores were spatially resolved by digital imaging with the Ca2+ indicator fura-2 in frog sympathetic neurons. Electrical stimulation evoked a rise in [Ca2+]i spreading radially from the periphery to the center of the ... More
Quantitative analysis of calcium (Ca2+) mobilization after stimulation with mitogens or anti-CD3 antibodies. Simultaneous fluo-3 and immunofluorescence flow cytometry.
AuthorsSei Y, Arora PK
JournalJ Immunol Methods
PubMed ID1672879
'A method is described for flow microfluorometric analysis of calcium mobilization in immune cells. This method is based on dual-color analysis of heterogeneous cell populations using fluo-3 and phycoerythrin (PE)-conjugated monoclonal antibody (mAb). This technique allows the detection of fluo-3 fluorescence as a measure of an increase in cytoplasmic free ... More
Influence of 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8) on cell cycle progression and proliferation of cultured arterial smooth muscle cells.
AuthorsDesgranges C, Campan M, Gadeau AP, Guerineau N, Mollard P, Razaka G
JournalBiochem Pharmacol
PubMed ID2009073
'8-(N,N-Diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8), a putative inhibitor of intracellular calcium mobilization, causes a dose-dependent inhibition of serum-induced proliferation of arterial smooth muscle cells in culture. Neither early rise in cytosolic calcium concentration nor induction of early induced cell cycle dependent genes (c-fos, ornithine decarboxylase) are inhibited after serum stimulation in presence of ... More
Free and bound sodium in pancreatic beta-cells exposed to glucose and tolbutamide.
AuthorsAli L, Grapengiesser E, Gylfe E, Hellman B, Lund PE
JournalBiochem Biophys Res Commun
PubMed ID2679551
'The effects of glucose and tolbutamide on the sodium handling of the pancreatic beta-cells were evaluated by measuring the total sodium content in intact islets from ob/ob-mice by integrating flame photometry and the free ion in individual beta-cells by dual wavelength fluorometry. Whereas increasing the glucose concentration from 3 to ... More
Visualization of cytosolic free calcium distribution and mobilization in guinea pig gastric chief cells.
AuthorsTsunoda Y, Yodozawa S, Tashiro Y
JournalCell Struct Funct
PubMed ID3224381
'Distribution and temporal change of free calcium concentration [( Ca2+]i) in single guinea pig gastric chief cells were visualized by a digital imaging microscope equipped with a microspectrofluorometer. The distribution was not homogeneous; a higher [Ca2+]i area was often localized in some restricted regions of the endoplasm and also at ... More
Changes in axon fluorescence during activity: molecular probes of membrane potential.
AuthorsCohen LB, Salzberg BM, Davila HV, Ross WN, Landowne D, Waggoner AS, Wang CH
JournalJ Membr Biol
PubMed ID4431037
In vitro release characteristics of diclofenac and hydrocortisone from Pluronic F-127 gels.
AuthorsTomida H, Shinohara M, Kuwada N, Kiryu S
JournalActa Pharm Suec
PubMed ID3450192
Intracellular ion activities and membrane transport in parietal cells measured with fluorescent dyes.
AuthorsNegulescu PA, Machen TE
JournalMethods Enzymol
PubMed ID2074799
The purpose of this chapter is to summarize the methods we have usedfor measuring intracellular pH with BCECF, intracellular calcium with fura-2 and intracellular sodium with SBFI in single, identified cells of gastricglands isolated from rabbits. ... More
Norepinephrine and serotonin increase the vasoconstrictor response of the perfused rat tail artery to changes in cytosolic Ca2+.
AuthorsThorin-Trescases N, Oster L, Atkinson J, Capdeville C
JournalEur J Pharmacol
PubMed ID2364996
Vasoconstriction and changes in cytosolic free Ca2+ concentrations (fura 2 with dual wavelength excitation) were measured simultaneously in the perfused tail artery of the rat. Vasoconstrictors (norepinephrine and 5-HT) appear to modulate contraction not only by changing the cytosolic free Ca2+ concentration but also by modifying the sensitivity of the ... More
Rapid kinetics of depolarization-induced changes in intrasynaptosomal calcium concentrations.
AuthorsTareilus E, Breer H
JournalNeurochem Int
PubMed ID1304325
The rapid kinetics of depolarization-evoked calcium influxes in isolated nerve terminals from rat cortex were monitored by stopped-flow spectrofluorimetry using specific indicators (Fluo-3, Indo-1). A very rapid increase in the intrasynaptosomal Ca(2+)-level was detected within the subsecond time range after depolarizing synaptosomes by mixing with physiological saline containing elevated K(+)-concentrations. ... More
Limited accumulation of cyclic AMP underlies a modest vasoactive-intestinal-peptide-mediated increase in cytosolic [Ca2+] transients in GH3 pituitary cells.
AuthorsMollard P, Zhang Y, Rodman D, Cooper DM
JournalBiochem J
PubMed ID1377907
The 4-chlorophenylthio analogue of cyclic AMP evoked profound and long-lasting changes in cytosolic [Ca2+] ([Ca2+]i) in pituitary-derived GH3 cells. However, vasoactive intestinal peptide (VIP), a hormone considered to act via cyclic AMP, was ineffective in modulating [Ca2+]i. The ability of VIP to modulate [Ca2+]i was enhanced by treatments that increased ... More
Synchronous free Ca2+ changes in individual neutrophils stimulated by leukotriene B4.
AuthorsDavies EV, Campbell AK, Hallett MB
JournalFEBS Lett
PubMed ID1657634
Calcium (Ca2+) signals were monitored in individual neutrophils using ratio imaging of fura-2. In contrast to N-formyl-L-leucyl-L-phenylalanine (f-met-leu-phe), which produced grossly asynchronous Ca2+ signals with delays in response (up to 60 s), leukotriene B4 (LTB4) provoked synchronous and immediate elevations in cytosolic free Ca2+. Some individual neutrophils which responded immediately ... More
Depletion of intracellular calcium stores by calcium ionophore A23187 induces the genes for glucose-regulated proteins in hamster fibroblasts.
AuthorsDrummond IA, Lee AS, Resendez E, Steinhardt RA
JournalJ Biol Chem
PubMed ID3114264
Animal cells respond to calcium ionophore (A23187) treatment with the coordinate induction of a set of genes encoding proteins identical to the glucose-regulated proteins (GRPs). By monitoring the intracellular free calcium with the fluorescent indicator fura-2 while employing both intracellular and extracellular calcium buffers, we demonstrated that A23187 can induce ... More
Intracellular ATP mimics GTP-gamma-S in generating Ca2+ oscillations in pancreatic beta-cells.
AuthorsLund PE, Grapengiesser E, Gylfe E, Hellman B
JournalBiochem Biophys Res Commun
PubMed ID2049100
Intracellular free calcium ([Ca2+]i) was measured in individual pancreatic beta-cells from mice using dual emission microfluorometry and the indicator Indo-1 applied by a patch clamp pipette. GTP-gamma-S (100 microM) injected together with 0.3 or 3 mM ATP evoked repetitive [Ca2+]i transients with a frequency of about 1 per min in ... More
Glutamate-induced increases in intracellular Ca2+ in cultured frog tectal cells mediated by direct activation of NMDA receptor channels.
AuthorsCline HT, Tsien RW
JournalNeuron
PubMed ID1704244
Influx of Ca2+ through NMDA channels may initiate the stabilization of coactive synapses during development of the retinotectal projection in frogs. Ca2+ imaging techniques were applied to cultured tectal cells to investigate whether excitatory amino acids cause a rise in [Ca2+]i. High [K+], NMDA, and glutamate increase [Ca2+]i in about ... More
Calcium rises abruptly and briefly throughout the cell at the onset of anaphase.
AuthorsPoenie M, Alderton J, Steinhardt R, Tsien R
JournalScience
PubMed ID3755550
Continuous measurement and imaging of the intracellular free calcium ion concentration ([Ca2+]i) of mitotic and interphase PtK1 cells was accomplished with the new fluorescent Ca2+ indicator fura-2. No statistically significant difference between basal [Ca2+]i of interphase and mitotic cells was detected. However, mitotic cells showed a rapid elevation of [Ca2+]i ... More
Rapid desensitization of substance P- but not carbachol-induced increases in inositol trisphosphate and intracellular Ca++ in rat parotid acinar cells.
AuthorsMcMillian MK, Soltoff SP, Talamo BR
JournalBiochem Biophys Res Commun
PubMed ID2446604
The effects of supramaximal concentrations of substance P and the cholinergic agonist carbachol on the accumulation of inositol trisphosphate and the elevation of the intracellular free calcium concentration were compared in rat parotid acinar cells. Substance P was fully as effective as carbachol at initial times, but there was a ... More
Quin2 protects against neuronal cell death due to Ca2+ overload.
AuthorsKudo Y, Takeda K, Yamazaki K
JournalBrain Res
PubMed ID1978795
Quin2-acetoxymethylester (quin2/AM) (50 microM), administered directly to the motoneuronal pool of the frog spinal cord, could be loaded into the motoneuron as well as the other cells in the lumbar region. Depolarizing responses of the ventral root to L-glutamate in the quin2-loaded side persisted even after prolonged exposure to A23187 ... More
Carbachol and KCl-induced changes in intracellular free calcium concentration in isolated, fura-2 loaded smooth-muscle cells from the anterior byssus retractor muscle of Mytilus edulis.
AuthorsIshii N, Simpson AW, Ashley CC
JournalBiochem Biophys Res Commun
PubMed ID3382397
Intracellular free calcium concentration [( Ca2+]1) was measured in suspensions of fura-2 loaded smooth-muscle cells isolated from the anterior byssus retractor muscle of Mytilus edulis. Successive application of 5mM carbachol (CCh) and 100mM KCl to the cells transiently elevated [Ca2+]1 from the resting value of 124 +/- 4.5nM (mean +/- ... More
Thimerosal causes calcium oscillations and sensitizes calcium-induced calcium release in unfertilized hamster eggs.
AuthorsSwann K
JournalFEBS Lett
PubMed ID1991508
Calcium-induced-calcium-release (CICR) was assayed in unfertilized golden hamster eggs by injecting Ca2+ and monitoring Ca2(+)-dependent hyperpolarizing responses (HRs) and Ca2(+)-sensitive fluo-3 fluorescence. Incubating eggs in the sulfhydryl reagent thimerosal caused [Ca2+]i oscillations as monitored by Ca2(+)-dependent HRs and decreased approximately 10-fold the Ca2+ injection current required to generate an HR ... More
Quantitation of lymphocyte intracellular free calcium signals using indo-1.
AuthorsOwen CS
JournalCell Calcium
PubMed ID3416353
The calcium-responsive fluorescent dye indo-1 has been used in lymphocyte suspensions to measure changes in internal free calcium concentration, [Ca2+]i, in response to crosslinking of cell surface immunoglobulin. The quantitation of [Ca2+]i requires that indo-am ester used to load the cells be completely hydrolyzed to the indo-1 form inside the ... More
Detection of zinc translocation into apical dendrite of CA1 pyramidal neuron after electrical stimulation.
AuthorsSuh SW,
JournalJ Neurosci Methods
PubMed ID18929598
Translocation of the endogenous cation zinc from presynaptic terminals to postsynaptic neurons after brain insult has been implicated as a potential neurotoxic event. Several studies have previously demonstrated that a brief electrical stimulation is sufficient to induce the translocation of zinc from presynaptic vesicles into the cytoplasm (soma) of postsynaptic ... More
Pluronic additives: a solution to sticky problems in digital microfluidics.
AuthorsLuk VN, Mo GCh, Wheeler AR,
JournalLangmuir
PubMed ID18481875
Digital microfluidics (DMF) is a promising technique for carrying out miniaturized, automated biochemical assays in which discrete droplets of reagents are actuated on the surface of an array of electrodes. A limitation for DMF is nonspecific protein adsorption to device surfaces, which interferes with assay fidelity and can cause droplets ... More
Pluronic F-127 affects the regulation of cytoplasmic Ca2+ in neuronal cells.
AuthorsSutachan JJ, Montoya G JV, Xu F, Chen D, Blanck TJ, Recio-Pinto E,
JournalBrain Res
PubMed ID16387285
Fura-2 is one of the most widely used cytoplasmic Ca2+ ([Ca2+]cyt) sensors. In studies using isolated dorsal root ganglion (DRG) neurons, the loading of Fura-2 AM is often facilitated by the use of pluronic F-127. In preliminary studies, we detected that the use of pluronic F-127 appeared to be affecting ... More
Effects of endothelin on intracellular Ca2+ and contractility in single ventricular myocytes from the ferret and human.
AuthorsQiu Z, Wang J, Perreault CL, Meuse AJ, Grossman W, Morgan JP
JournalEur J Pharmacol
PubMed ID1516644
We investigated the role of endothelin-1 on peak intracellular Ca2+ ([Ca2+]i) and peak shortening of ventricular myocytes (loaded with indo-1/AM) from failing human hearts. 10 nM of ET-1 significantly increased the cell peak shortening (84 +/- 29%, P less than 0.05) without significantly increasing the peak [Ca2+]i (15 +/- 7%, ... More
Ethanol inhibits mitogen-induced calcium mobilization in mouse splenocytes.
AuthorsSei Y, McIntyre T, Skolnick P, Arora PK
JournalLife Sci
PubMed ID1346466
Ethanol inhibited the mitogen-induced initial increase in cytoplasmic free-calcium [Ca2+]i in mouse splenocytes. This effect was concentration-dependent, reversible, and observed at pharmacologically relevant concentrations (24-166mM). Other short-chain alcohols such as propanol, butanol, and pentanol also inhibited this mitogen-induced increase in [Ca2+]i. The potencies of these alcohols to produce this effect ... More
Non-calcium environmental sensitivity of intracellular Indo-1.
AuthorsOwen CS, Sykes NL, Shuler RL, Ost D
JournalAnal Biochem
PubMed ID2048716
Several mechanisms might account for the difference between spectra of Indo-1 measured on intracellular and on extracellular dye. Experiments to discriminate between various possibilities used quantitative analyses of intracellular and extracellular Indo-1 spectra. Intracellular calcium-free dye showed a 20-nm blue shift, when compared with the extracellular case. However, the calcium-bound ... More
Acidic amino acids evoke a smaller [Ca2+]i rise in GABAergic than non-GABAergic hippocampal neurons.
AuthorsSegal M, Greenberger V
JournalNeurosci Lett
PubMed ID1380146
Changes in intracellular calcium concentration ([Ca2+]i) in response to topical application of the glutamate agonists N-methyl-D-aspartate (NMDA), or amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) were measured in cultured rat hippocampal neurons loaded with Fluo-3 and visualized in a confocal laser scanning microscope. These neurons were subsequently stained for the presence of the enzyme ... More
Mitogen-induced early increase in cytosolic free Mg2+ concentration in single Swiss 3T3 fibroblasts.
AuthorsIshijima S, Sonoda T, Tatibana M
JournalAm J Physiol
PubMed ID1767812
Events related to the early mobilization of Mg2+ in mammalian cells in response to external stimuli are not well characterized. We examined changes in cytoplasmic free Mg2+ concentrations ([Mg2+]i) after mitogenic stimulation in single mouse Swiss 3T3 fibroblasts, using digital ratio imaging microscopy of the fluorescent probe mag-fura-2. Stimulation with ... More
Transmitter release increases intracellular calcium in perisynaptic Schwann cells in situ.
AuthorsJahromi BS, Robitaille R, Charlton MP
JournalNeuron
PubMed ID1351731
Glial cells isolated from the nervous system are sensitive to neurotransmitters and may therefore be involved in synaptic transmission. The sensitivity of individual perisynaptic Schwann cells to activity of a single synapse was investigated, in situ, at the frog neuromuscular junction by monitoring changes in intracellular Ca2+ in the Schwann ... More
Identification of orphan G protein-coupled receptor ligands using FLIPR assays.
AuthorsRobas NM, Fidock MD,
JournalMethods Mol Biol
PubMed ID15867462
G protein-coupled receptors (GPCRs) make up the largest and most diverse family of transmembrane proteins and respond to a wide variety of stimuli including biogenic amines, peptides, bioactive lipids, hormones, and light. Agonist binding to these receptors activates intracellular signalling events mediated by G proteins, such as modulation of intracellular ... More
Measurement of cytoplasmic calcium in single microvessels with increased permeability.
AuthorsHe P, Pagakis SN, Curry FE
JournalAm J Physiol
PubMed ID2337172
We investigated the hypothesis that an increase in cytoplasmic calcium ion concentration, [Ca2+]i, is one of the mechanisms responsible for increased microvessel permeability. We loaded the cells forming the walls of individually perfused microvessels in frog mesentery with fura-2 and measured [Ca2+]i in the control state and after adding the ... More
Continuous fluorimetric assessment of the changes in cytoplasmic calcium concentration during exposure of rat isolated myocardium to conditions of simulated ischaemia.
AuthorsNorthover BJ
JournalBr J Pharmacol
PubMed ID2390672
1. The cytoplasmic calcium concentration ([Ca]c) of rat isolated atrial myocardium was assessed with the dye indo-1. Dye-loaded atria were superfused with physiological salt solution and excited with radiation at 360 nm, while epifluorescence emissions were collected simultaneously at 400 nm and 500 nm. The ratio of these emissions was ... More
Biphasic increase in intracellular calcium induced by platelet-activating factor in macrophages.
AuthorsRandriamampita C, Trautmann A
JournalFEBS Lett
PubMed ID2544447
In single mouse macrophages stimulated by platelet-activating factor (PAF), the intracellular calcium concentration (Cai) monitored with fura-2 at room temperature presents a biphasic increase, including a transient and a more sustained component. After pulse administration of PAF, the first phase lasts for a few seconds and reaches a peak value ... More
Interleukin 1 beta increases the cytosolic free sodium concentration in isolated rat islets of Langerhans.
Interleukin 1 (IL-1) exerts both stimulatory and inhibitory (cytotoxic) effects on insulin-producing beta cells in isolated pancreatic islets. Since alteration in ion fluxes is crucial for endocrine cell activation and is a denominator of cell death, and since IL-1 was recently shown to increase the total sodium content in a ... More
Regulation of intracellular free calcium in normal and dystrophic mouse cerebellar neurons.
AuthorsHopf FW, Steinhardt RA
JournalBrain Res
PubMed ID1511288
We measured free intracellular calcium ([Ca2+]i) in cultured cerebellar granule cells from normal and mdx mice. Resting levels of ([Ca2+]i) were 24% higher in the dystrophic neurons (normal: 61.2 +/- 1.5 nM calcium, n = 104; dystrophic: 76.1 +/- 2.4 nM calcium, n = 136, P less than 0.01). Dystrophic ... More
Two-photon excitation imaging of pancreatic islets with various fluorescent probes.
AuthorsTakahashi N, Nemoto T, Kimura R, Tachikawa A, Miwa A, Okado H, Miyashita Y, Iino M, Kadowaki T, Kasai H
JournalDiabetes
PubMed ID11815453
Various fluorescent probes were assessed for investigating intact islets of Langerhans using two-photon excitation imaging. Polar fluorescent tracers applied on the outside rapidly (within 3 min) penetrated deep into the islets via microvessels. Likewise, an adenovirus carrying a Ca(2+)-sensitive green fluorescent protein mutant gene, yellow cameleon 2.1, was successfully transfected ... More
Transmembrane gradients of free Na+ in isolated heart mitochondria estimated using a fluorescent probe.
AuthorsJung DW, Apel LM, Brierley GP
JournalAm J Physiol
PubMed ID1566810
The concentration of free Na+ in the matrix of isolated pig heart mitochondria has been monitored using the fluorescent probe sodium-binding benzofuran isophthalate (SBFI) developed by Minta and Tsien (J. Biol. Chem. 264: 19449-19457, 1989). SBFI was sequestered in the matrix by hydrolysis of the permeant acetoxymethyl ester. The sequestered ... More
Mechanisms of vasopressin-induced increase in intracellular Na+ in vascular smooth muscle cells.
AuthorsOkada K, Ishikawa S, Saito T
JournalAm J Physiol
PubMed ID1661077
The present study was undertaken to examine the interrelationship between the arginine vasopressin (AVP)-induced dynamic changes in intracellular Ca2+ and Na+ concentrations in cultured rat aortic vascular smooth muscle cells (VSMC) by the direct measurements of intracellular Na+ concentration [( Na+]i), cytosolic free calcium [( Ca2+]i), and intracellular pH (pHi) ... More
Desensitization to norepinephrine includes refractoriness of calcium release in myocardial cells.
AuthorsErne P, Hermsmeyer K
JournalBiochem Biophys Res Commun
PubMed ID3348780
Localization of myoplasmic free calcium was measured in fura2-loaded single rat myocardial cells to determine whether the mechanism of norepinephrine desensitization includes redistribution of calcium. Fluorescence intensities at each pixel were quantitated by use of a photon-counting, microchannel plate camera. From these images, values of calcium-dependent fluorescence intensity averages in ... More
Fura-2 fluorescence is localized to mitochondria in endothelial cells.
AuthorsSteinberg SF, Bilezikian JP, Al-Awqati Q
JournalAm J Physiol
PubMed ID3688220
The new, highly fluorescent, calcium-sensitive dye, fura-2, can be loaded nondisruptively into intact cells by means of its permeant ester and used to measure the free calcium ion concentration in individual cells. For fura-2 to signal cytosolic calcium, it must be distributed homogeneously and exclusively throughout the cytoplasmic space. However, ... More
Histamine receptors in human fibroblasts: inositol phosphates, Ca2+, and cell growth.
AuthorsJohnson CL, Johnson CG, Bazan E, Garver D, Gruenstein E, Ahluwalia M
JournalAm J Physiol
PubMed ID2316639
Histamine stimulated inositol phosphate formation by human skin fibroblasts. The effect of histamine was reduced but still readily apparent in the absence of extracellular Ca2+. Histamine caused a transient increase in intracellular free Ca2+ as detected by indo-1 and fura-2 fluorescence studies on cell populations and on individual cells. Similar ... More
Intracellular free calcium concentration measured with 19F NMR spectroscopy in intact ferret hearts.
AuthorsMarban E, Kitakaze M, Kusuoka H, Porterfield JK, Yue DT, Chacko VP
JournalProc Natl Acad Sci U S A
PubMed ID3112778
Changes in the intracellular free Ca2+ concentration, [Ca2+]i, mediate excitation-contraction coupling in the heart and contribute to cellular injury during ischemia and reperfusion. To study these processes directly, we measured [Ca2+]i in perfused ferret (Mustela putorius furo) hearts using 19F NMR spectroscopy to detect the 5,5'-difluoro derivative of the Ca2+ ... More
Intracellular endocardial calcium and myocardial function in rat hearts.
AuthorsWikman-Coffelt J, Wu ST, Parmley WW
JournalCell Calcium
PubMed ID1849795
Analyses of [Ca2+]i in the isolated beating rat heart (using Indo-1 dye) demonstrated a direct relationship between developed pressure and each of the following: (a) systolic [Ca2+]i; (b) amplitude of [Ca2+]i transients; and (c) diastolic level of [Ca2+]i. Agents which increased cAMP levels, augmented amplitude of [Ca2+]i transients, and lowered ... More
Nitroglycerin relaxes canine coronary arterial smooth muscle without reducing intracellular Ca2+ concentrations measured with fura-2.
AuthorsYanagisawa T, Kawada M, Taira N
JournalBr J Pharmacol
PubMed ID2510898
1. Changes in cytoplasmic Ca2+ concentration ([Ca2+]1) were measured simultaneously with force by a microfluorometric method using a calcium indicator, fura-2, in canine coronary arterial smooth muscle cells. 2. Depolarization by high (30-90 mM) KCl-physiological salt solution (PSS) produced concentration-dependent increases in force and [Ca2+]i. 3. The KCl-induced increase in ... More
In situ calibration of fura-2 and BCECF fluorescence in adult rat ventricular myocytes.
AuthorsBorzak S, Kelly RA, Krämer BK, Matoba Y, Marsh JD, Reers M
JournalAm J Physiol
PubMed ID2396701
Quantitation of Ca+ and H+ activities within cells using presently available fluorescent probes is optimal when the fluorescence signal is calibrated in situ after each experiment. Fura-2 and 2',7'-bis(2-carboxy-ethyl)-5,6-carboxyfluoroscein (BCECF) are difficult to calibrate in freshly dissociated adult cardiac myocytes because calibration procedures produce cellular hypercontracture. In situ calibration was ... More
Relationship between force and Ca2+ in anococcygeal and vas deferens smooth muscle cells of the mouse.
AuthorsBoland B, Himpens B, Gillis JM, Casteels R
JournalPflugers Arch
PubMed ID1630884
We compared the changes of the cytoplasmic Ca2+ concentration ([Ca2+]i), as measured with the fluorescent Ca2+ indicator fura-2, and the force development in intact smooth muscle of the tonic anococcygeus (AC) and the phasic vas deferens (VD) of the mouse, during activation by K+ depolarization and by agonists. Resting [Ca2+]i ... More
Failure of thapsigargin to alter ion transport in human sweat gland epithelia while intracellular Ca2+ concentration is raised.
AuthorsPickles RJ, Cuthbert AW
JournalJ Biol Chem
PubMed ID1321817
Cai in cultured human sweat gland epithelial monolayers was measured using Fura-2 fluorescence. Thapsigargin (Tg) caused a sustained increase in Cai, the rate of rise being slower but the magnitude greater than with the agonists lysylbradykinin and ATP. Tg caused an irreversible change such that even after it was removed ... More
Repetitive calcium transients and the role of calcium in exocytosis and cell cycle activation in the mouse egg.
AuthorsKline D, Kline JT
JournalDev Biol
PubMed ID1728596
The role of calcium in cortical granule exocytosis and activation of the cell cycle at fertilization was examined in the mouse egg using the calcium chelator BAPTA (1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid) and the fluorescent calcium indicator fluo-3. BAPTA and fluo-3 were introduced into zona-free mouse eggs by a 30-min incubation with 0.01-50 ... More
Flow cytometric measurement of cytoplasmic free calcium in human peripheral blood T lymphocytes with fluo-3, a new fluorescent calcium indicator.
AuthorsVandenberghe PA, Ceuppens JL
JournalJ Immunol Methods
PubMed ID2107260
Cytoplasmic free calcium [( Ca2+]i) is a key intracellular messenger in many cell types. We have used fluo-3, a recently developed calcium probe, to study [Ca2+]i in resting and stimulated human peripheral blood T lymphocytes. The spectral properties of fluo-3 permit analysis of [Ca2+]i in flow cytometers with a 488 ... More
Inhibition of tyrosine phosphorylation prevents thrombin-induced mitogenesis, but not intracellular free calcium release, in vascular smooth muscle cells.
AuthorsWeiss RH, Nuccitelli R
JournalJ Biol Chem
PubMed ID1544934
alpha-Thrombin, a G-protein-coupled receptor agonist, is mitogenic for neonatal vascular smooth muscle (VSM) cells, but it also causes secretion of the tyrosine kinase-coupled receptor agonist platelet-derived growth factor (PDGF). In order to determine the role of growth factors with tyrosine kinase-coupled receptors in thrombin's mitogenic signal transduction cascade, the synergistic ... More
The effect of endotoxemia on concanavalin A induced alterations in cytoplasmic free calcium in rat spleen cells as determined with Fluo-3.
AuthorsHagar AF, Spitzer JA
JournalCell Calcium
PubMed ID1633610
The effects of acute (3 h) and chronic (30 h) in vivo infusions of Escherichia coli endotoxin on the Ca2+ homeostasis of rat spleen cells was investigated. Conditions were established for obtaining reliable estimates of [Ca2+]i in these cells using the newly-developed Ca2+ indicator Fluo-3. The resting [Ca2+]i of splenocytes ... More
Calcium oscillations in freshly dispersed and cultured interstitial cells from canine colon.
AuthorsPublicover NG, Horowitz NN, Sanders KM
JournalAm J Physiol
PubMed ID1550205
Spontaneous oscillations in intracellular Ca2+ concentration ([Ca2+]i) and membrane potential were used to monitor rhythmicity in freshly dispersed and cultured interstitial cells (IC) from the canine colon. The frequency of oscillations and responses to a number of channel blockers, agonists, ionic substitutions, and temperature were similar in freshly dispersed and ... More
Uptake of sodium in protoplasts of salt-sensitive and salt-tolerant cultivars of rice, Oryza sativa L. determined by the fluorescent dye SBFI.
AuthorsKader MA, Lindberg S
JournalJ Exp Bot
PubMed ID16275670
In this study, the uptake of Na+ into the cytosol of rice (Oryza sativa L. cvs Pokkali and BRRI Dhan29) protoplasts was measured using the acetoxy methyl ester of the fluorescent sodium-binding benzofuran isopthalate, SBFI-AM, and fluorescence microscopy. By means of inhibitor analyses the mechanisms for uptake and sequestration of ... More