Arabidopsis abi1-1 and abi2-1 phosphatase mutations reduce abscisic acid-induced cytoplasmic calcium rises in guard cells.
AuthorsAllen GJ,Kuchitsu K,Chu SP,Murata Y,Schroeder JI
JournalThe Plant cell
PubMed ID10488243
Elevations in cytoplasmic calcium ([Ca(2)+](cyt)) are an important component of early abscisic acid (ABA) signal transduction. To determine whether defined mutations in ABA signal transduction affect [Ca(2)+](cyt) signaling, the Ca(2)+-sensitive fluorescent dye fura 2 was loaded into the cytoplasm of Arabidopsis guard cells. Oscillations in [Ca(2)+](cyt) could be induced when ... More
CD5 antibodies increase intracellular ionized calcium concentration in T cells.
AuthorsJune CH,Rabinovitch PS,Ledbetter JA
JournalJournal of immunology (Baltimore, Md. : 1950)
PubMed ID3106489
The binding of a variety of monoclonal antibodies to the CD5 (T, gp67) pan T cell differentiation antigen has been shown to potentiate T cell proliferation. In this paper we show that CD5 monoclonal antibodies cause increased intracellular free calcium concentration ([Ca2+]i) in T cells. An increase in [Ca2+]i occurred ... More
Fluorescent indicators of ion concentrations.
AuthorsTsien RY
JournalMethods Cell Biol
PubMed ID2538708
Detection of La3+ influx in ventricular cells by indo-1 fluorescence.
AuthorsPeeters GA, Kohmoto O, Barry WH
JournalAm J Physiol
PubMed ID2919662
'We exposed indo-1-loaded cultured embryonic chick ventricular cells to 0.03-1.0 mM extracellular lanthanum concentration ([La3+]o) and simultaneously measured cell contractile motion and the 410/480 nm fluorescence intensity ratio. After exposure to La3+, ventricular cells stopped contracting and relaxed within seconds, and the 410/480 fluorescence ratio increased. The increase in the ... More
Mitochondrial control of calcium-channel gating: a mechanism for sustained signaling and transcriptional activation in T lymphocytes.
AuthorsHoth M, Button DC, Lewis RS
JournalProc Natl Acad Sci U S A
PubMed ID10973476
'In addition to their well-known functions in cellular energy transduction, mitochondria play an important role in modulating the amplitude and time course of intracellular Ca(2+) signals. In many cells, mitochondria act as Ca(2+) buffers by taking up and releasing Ca(2+), but this simple buffering action by itself often cannot explain ... More
CD19 monoclonal antibody HD37 inhibits anti-immunoglobulin-induced B cell activation and proliferation.
AuthorsPezzutto A, Dörken B, Rabinovitch PS, Ledbetter JA, Moldenhauer G, Clark EA
JournalJ Immunol
PubMed ID2437199
'The 95 Kd CD19 antigen is the broadest lineage specific surface marker for B cells: it is present on the surface of virtually all B lymphocytes, including early B progenitor cells. In this study we have evaluated the function of the CD19 antigen by using the CD19 mAb HD37. Binding ... More
Intracellular free Ca2+ in the cell cycle in human fibroblasts: transitions between G1 and G0 and progression into S phase.
AuthorsWahl M, Gruenstein E
JournalMol Biol Cell
PubMed ID8485320
'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 [Ca2+]i (Rabinovitch et al., 1986; Vincentini and Villereal, 1986; Hesketh et ... More
Lipid rafts, the sarcoplasmic reticulum and uterine calcium signalling: an integrated approach.
AuthorsNoble K, Zhang J, Wray S
JournalJ Physiol
PubMed ID16239270
'The pathways involved in Ca2+ signalling in the uterus remain incompletely understood, impairing our ability to prevent preterm and difficult labours. In this review we focus on two elements in the pathway of Ca2+ signalling that have recently emerged as playing important roles: membrane lipid rafts and the sarcoplasmic reticulum. ... More
Calcium permeability of the neuronal nuclear envelope: evaluation using confocal volumes and intracellular perfusion.
AuthorsO'Malley DM
JournalJ Neurosci
PubMed ID7931542
'In many calcium-imaging studies, the nuclear envelope appears to maintain a gradient of free calcium between the nucleus and cytosol. This issue was examined by loading amphibian sympathetic neurons with the calcium indicator fluo 3 via whole-cell patch clamping. Confocal optical sectioning allowed acquisition of independent calibration curves for the ... More
Protein and acidosis alter calcium-binding and fluorescence spectra of the calcium indicator indo-1.
AuthorsBaker AJ, Brandes R, Schreur JH, Camacho SA, Weiner MW
JournalBiophys J
PubMed ID7819496
'The fluorescent indicator indo-1 is widely used to monitor intracellular calcium concentration. However, quantitation is limited by uncertain effects of the intracellular environment on indicator properties. The goal of this study was to determine the effects of protein and acidosis on the fluorescence spectra and calcium dissociation constant (Kd) of ... More
Simultaneous recording of Indo-1 fluorescence and Na+/Ca2+ exchange current reveals two components of Ca2(+)-release from sarcoplasmic reticulum of cardiac atrial myocytes.
AuthorsLipp P, Pott L, Callewaert G, Carmeliet E
JournalFEBS Lett
PubMed ID2261988
'Simultaneous measurements of intracellular Ca2(+)-concentration ([Ca2+]i) using Indo-1 and the current generated by electrogenic Na+/Ca2(+)-exchange (INaCa) have been performed on atrial myocytes from hearts of adult guinea-pigs. Whereas the fluorescence-measurements provide information on global [Ca2+]i, InaCa which is a linear function of Ca2(+)-concentration, indicates subsarcolemmal [Ca2+]. Under conditions in which ... More
Calcium signalling in non-excitable cells: notes on oscillations and store refilling.
AuthorsRink TJ, Hallam TJ
JournalCell Calcium
PubMed ID2670242
'Technical advances in studying cellular calcium concentrations, and discoveries about many aspects of signal transduction have transformed this field of biology since this Journal was launched a decade ago. At that time monitoring of the key variable, cytoplasmic free Ca2+ concentration [Ca2+]i, was possible with aequorin or arsenazo ill mainly ... More
Extracellular nucleotides mediate Ca2+ fluxes in J774 macrophages by two distinct mechanisms.
AuthorsGreenberg S, Di Virgilio F, Steinberg TH, Silverstein SC
JournalJ Biol Chem
PubMed ID3392016
'We have studied the effects of extracellular nucleotides on the cytosolic free calcium concentration [( Ca2+]i) in J774 macrophages using quin2 and indo-1 as indicator dyes. Micromolar quantities of ATP induced a biphasic increase in [Ca2+]i: a rapid and transient increase (peak I) which was due to mobilization of Ca2+ ... More
Differences in regulation between nuclear and cytoplasmic Ca2+ in cultured smooth muscle cells.
AuthorsHimpens B, De Smedt H, Droogmans G, Casteels R
JournalAm J Physiol
PubMed ID1386189
'The free Ca2+ concentrations in the nucleus ([Ca2+]n) and cytoplasm ([Ca2+]c) of cultured smooth muscle cells were estimated using the fluorescent dye indo-1 and the ACAS 570 confocal laser microscope. In resting DDT1MF2 smooth muscle cells [Ca2+]n was found to be lower than [Ca2+]c. Both values increased transiently in response ... More
Intracellular [Ca2+] and inositol phosphates in avian nasal gland cells.
AuthorsShuttleworth TJ, Thompson JL
JournalAm J Physiol
PubMed ID2596581
'Isolated cells from the nasal salt gland of ducklings (Anas platyrhynchos) were evaluated as a model system for the study of the muscarinic activation of exocrine ion secretion. Cells loaded with the fluorescent probe indo-1 were used to study changes in intracellular Ca2+ concentration [( Ca2+]i) after stimulation. Changes in ... More
Relation of mitochondrial and cytosolic free calcium to cardiac myocyte recovery after exposure to anoxia.
'Mitochondrial calcium overload has been suggested as a marker for irreversible injury in the ischemic heart. A new technique is used to measure dynamic changes in mitochondrial free calcium concentration ([Ca2+]m) in electrically stimulated (0.2 Hz) adult rat cardiac myocytes during exposure to anoxia and reoxygenation. Cells were incubated with ... More
Modulation of L-type Ca2+ current by fast and slow Ca2+ buffering in guinea pig ventricular cardiomyocytes.
AuthorsYou Y, Pelzer DJ, Pelzer S
JournalBiophys J
PubMed ID8994602
'Free Ca2+ near Ca2+ channel pores is expected to be lower in cardiomyocytes dialyzed with bis-(o-amino-phenoxy)-ethane-N,N,N'',N''-tetraacetic acid (BAPTA) than with ethyleneglycol-bis-(beta-aminoethyl)-N,N,N'',N''-tetraacetic acid (EGTA) because BAPTA chelates incoming Ca2+ more rapidly. The consequences of intracellular Ca2+ buffering by BAPTA (0.2-60 mM) and by EGTA (0.2-67 mM) on whole-cell L-type Ca2+ current ... More
Intracellular Ca2+ transients induced by high external K+ and tetracaine in cultured rat myotubes.
AuthorsJaimovich E, Rojas E
JournalCell Calcium
PubMed ID8033194
'Cultured myotubes from rat neonatal skeletal muscle were used to measure intracellular Ca2+ concentration ([Ca2+]i) and membrane potentials (Vm) using the Indo-1 microfluorimetry method and the nystatin perforated membrane patch technique, respectively. Sudden increases in external [K+]o from 5 mM to either 22, 42 or 84 mM elicited transient elevations ... More
Regulation of anaphase chromosome motion in Tradescantia stamen hair cells by calcium and related signaling agents.
AuthorsZhang DH, Callaham DA, Hepler PK
JournalJ Cell Biol
PubMed ID2114409
'Several lines of evidence support the idea that increases in the intracellular free calcium concentration [( Ca2+]i) regulate chromosome motion. To directly test this we have iontophoretically injected Ca2+ or related signaling agents into Tradescantia stamen hair cells during anaphase and measured their effect on chromosome motion and on the ... More
Multiwavelength method for measuring concentration of free cytosolic calcium using the fluorescent probe indo-1.
AuthorsPopov EG, Gavrilov YIu, Pozin EYa Gabbasov ZA
JournalArch Biochem Biophys
PubMed ID3341781
'It is assumed that the spectra of fluorescent probes indo-1 and fura-2 in the cytoplasm are linear combinations of the spectra of calcium-bound and free probes with weight factors proportional to the concentrations of these forms. When the concentration of calcium is measured by the dual-wavelength method, the above assumption ... More
A new generation of Ca2+ indicators with greatly improved fluorescence properties.
AuthorsGrynkiewicz G, Poenie M, Tsien RY
JournalJ Biol Chem
PubMed ID3838314
'A new family of highly fluorescent indicators has been synthesized for biochemical studies of the physiological role of cytosolic free Ca2+. The compounds combine an 8-coordinate tetracarboxylate chelating site with stilbene chromophores. Incorporation of the ethylenic linkage of the stilbene into a heterocyclic ring enhances the quantum efficiency and photochemical ... More
Protein kinase C interaction with calcium: a phospholipid-dependent process.
AuthorsBazzi MD, Nelsestuen GL
JournalBiochemistry
PubMed ID2271522
'The calcium-binding properties of calcium- and phospholipid-dependent protein kinase C (PKC) were investigated by equilibrium dialysis in the presence and the absence of phospholipids. Calcium binding to PKC displayed striking and unexpected behavior; the free proteins bound virtually no calcium at intracellular calcium concentrations and bound limited calcium (about 1 ... More
Investigation of factors affecting fluorometric quantitation of cytosolic [Ca2+] in perfused hearts.
AuthorsBrandes R, Figueredo VM, Camacho SA, Baker AJ, Weiner MW
JournalBiophys J
PubMed ID8298028
'The goal of these studies was to examine the effects of several factors that may artifactually influence quantitation of cytosolic [Ca2+], [Ca2+]c, while using the fluorescent calcium indicator Indo-1. The following factors were investigated: 1) a possible fluorescence contribution from unhydrolized Indo-1/AM (by Mn2+ quenching), 2) Ca2+ buffering by Indo-1 ... More
Regulation of calcium current by voltage and cytoplasmic calcium in canine gastric smooth muscle.
AuthorsVogalis F, Publicover NG, Sanders KM
JournalAm J Physiol
PubMed ID1312782
'The regulation of Ca2+ current by intracellular Ca2+ was studied in isolated myocytes from the circular layer of canine gastric antrum. Ca2+ current was measured with the whole cell patch-clamp technique, and changes in cytoplasmic Ca2+ ([Ca2+]i) were simultaneously measured with indo-1 fluorescence. Ca2+ currents were activated by depolarization and ... More
On the possibility of calcium imaging using Indo-1 with three-photon excitation.
AuthorsGryczynski I, Szmacinski H, Lakowicz JR
JournalPhotochem Photobiol
PubMed ID7480157
'We show that the calcium fluorophore Indo-1 can be excited by simultaneous absorption of three-photons at 885 nm, a wavelength readily available from Ti:sapphire lasers. Three-photon excitation was demonstrated by the emission intensity of Indo-1 which depended on the cube of the laser power, and by a higher anisotropy than ... More
Flow cytometric analysis of murine splenic B lymphocyte cytosolic free calcium response to anti-IgM and anti-IgD.
AuthorsChused TM, Wilson HA, Greenblatt D, Ishida Y, Edison LJ, Tsien RY, Finkelman FD
JournalCytometry
PubMed ID3497786
'The accurate measurement of ionized intracellular calcium [Ca++]i in single cells by flow cytometry with the use of a new fluorescent calcium chelator, indo-1, is described. We have developed a dependable in situ calibration technique that indicates a resting [Ca++]i in lymphocytes of 100 nM. The enhanced fluorescence of this ... More
Characterization of indo-1 and quin-2 as spectroscopic probes for Zn2(+)-protein interactions.
AuthorsJefferson JR, Hunt JB, Ginsburg A
JournalAnal Biochem
PubMed ID2116741
'1-[2-Amino-5-(6-carboxyindol-2-yl)phenoxyl]-2-(2''- amino-5''-methylphenoxy)ethane-N,N,N'',N''-tetraacetic acid (indo-1) and 2-[2-(bis(carboxymethyl)amino-5-methylphenoxy) methyl]-6- methyl-8-[bis-(carboxymethyl)amino]quinoline (quin-2) are sensitive, spectral indicators for Zn2+. Additions of subsaturating Zn2+ to 10-80 microM indo-1 or quin-2 at pH 7.0 produce uv difference spectra with isosbestic wavelengths at 342 and 282 nm or at 342, 317, and 252 nm, respectively. Formation of ... More
Decreased efficacy of inositol 1,4,5-trisphosphate to elicit calcium mobilization from cerebrocortical microsomes of aged rats.
AuthorsBurnett DM, Daniell LC, Zahniser NR
JournalMol Pharmacol
PubMed ID2157962
'The effects of aging on the ability of brain microsomes to sequester calcium in response to ATP stimulation and to release calcium in response to inositol 1,4,5-trisphosphate (IP3) stimulation were examined. Calcium uptake and release were compared in microsomal preparations from the cerebral cortex, hippocampus, thalamus, and cerebellum of 3-, ... More
Mitogen-induced changes in Ca2+ permeability are not mediated by voltage-gated K+ channels.
AuthorsGelfand EW, Cheung RK, Grinstein S
JournalJ Biol Chem
PubMed ID2427511
'Binding of mitogenic lectins to T lymphocytes results in elevated cytoplasmic Ca2+ concentrations ([Ca2+]i). This change in [Ca2+]i is thought to be essential for cellular proliferation. In addition, the lectins increase the conductance to K+ through voltage-sensitive channels. Based on the inhibitory effect of K+ channel blockers on lectin-induced mitogenesis, ... More
Intracellular calcium signals measured with indo-1 in isolated skeletal muscle fibres from control and mdx mice.
AuthorsCollet C, Allard B, Tourneur Y, Jacquemond V
JournalJ Physiol
PubMed ID10523411
'1. Intracellular free calcium concentration ([Ca2+]i) was measured with the fluorescent indicator indo-1 in single skeletal fibres enzymatically isolated from the flexor digitorum brevis and interosseus muscles of control and dystrophic mdx C57BL/10 mice. Measurements were taken from a portion of fibre that was voltage clamped to allow detection of ... More
Measurement of cytoplasmic calcium in aleurone protoplasts using indo-1 and fura-2.
AuthorsBush DS, Jones RL
JournalCell Calcium
PubMed ID3435914
'Previous attempts to measure cytoplasmic Ca2+ in plant cells using the new generation of fluorescent probes, indo-1 and fura-2, have been unsuccessful. We investigated the use of indo-1 and fura-2 to measure cytoplasmic Ca2+ in barley aleurone protoplasts and found that indo-1 could be successfully used when it was loaded ... More
ATP activates non-selective cation channels and calcium release in inner hair cells of the guinea-pig cochlea.
AuthorsSugasawa M, Erostegui C, Blanchet C, Dulon D
JournalJ Physiol
PubMed ID8815205
'1. ATP-evoked currents and Ca2+ signals were simultaneously recorded in isolated inner hair cells (IHC) of guinea-pig cochlea by combining conventional whole-cell or perforated patch clamp recording with indo-1 dual emission microfluorometry. 2. In most IHCs, voltage clamped near resting membrane potential (-40 mV), extracellular ATP evoked a rapid inward ... More
Angiotensin II inhibits calcium and M current channels in rat sympathetic neurons via G proteins.
AuthorsShapiro MS, Wollmuth LP, Hille B
JournalNeuron
PubMed ID7516687
'We characterized inhibition of N-type Ca2+ and M current K+ channels in rat superior cervical ganglion neurons by angiotensin II (angioII) using the patch clamp. Of 120 neurons, 97 showed inhibition of ICa (mean 32%), which was slow in onset and very slow to reverse under whole-cell recording conditions. This ... More
Calcium-induced calcium release in neurones.
AuthorsVerkhratsky A, Shmigol A
JournalCell Calcium
PubMed ID8653752
'Neurones express several subtypes of intracellular Ca2+ channels, which are regulated by cytoplasmic calcium concentration ([Ca2+]c) and provide the pathway for Ca(2+)-induced Ca2+ release (CICR) from endoplasmic reticulum Ca2+ stores. The initial studies of CICR which employed several pharmacological tools (and in particular caffeine and ryanodine) demonstrated that: (i) caffeine ... More
Induction of functionally active platelet-activating factor receptors in rat alveolar macrophages.
'Freshly isolated alveolar macrophages from control rats express low numbers of platelet-activating factor (PAF) receptors (4.6 x 10(3) binding sites/cell, Kd = 1.3 nM). These receptors appeared to be functionally inert in that PAF failed to induce intracellular calcium mobilization in the cells. Following brief exposure of rats to ozone ... More
Conversion of esterified fura-2 and indo-1 to Ca2+-sensitive forms by mitochondria.
AuthorsGunter TE, Restrepo D, Gunter KK
JournalAm J Physiol
PubMed ID3421312
'Rat liver mitochondria are shown to convert the acetoxymethyl ester forms of fura-2 and indo-1 into Ca2+-dependent forms of these indicators. The excitation spectrum of the Ca2+-dependent conversion product of fura-2 acetoxymethyl ester is shown to be similar to that of the pentacarboxylic acid form of fura-2. A systematic investigation ... More
Inositol trisphosphate and cyclic ADP-ribose-mediated release of Ca2+ from single isolated pancreatic zymogen granules.
'In pancreatic acinar cells low (physiological) agonist concentrations evoke cytosolic Ca2+ spikes specifically in the apical secretory pole that contains a high density of secretory (zymogen) granules (ZGs). Inositol 1,4,5-trisphosphate (IP3) is believed to release Ca2+ from the endoplasmic reticulum, but we have now tested whether the Ca(2+)-releasing messengers IP3 ... More
Measurement of CA2+ transients using simultaneous dual-emission microspectrofluorimetry and electrophysiology in individual pituitary cells.
AuthorsMollard P, Guerineau N, Audin J, Dufy B
JournalBiochem Biophys Res Commun
PubMed ID2590186
'Cytosolic free calcium concentration, [Ca2+]i, was monitored in individual pituitary GH3B6 cells, loaded with the fluorescent Ca2+ indicator indo 1 either by internal perfusion through a patch clamp pipette or by exposure to indo 1 acetoxymethyl ester, with the use of a dual-emission apparatus for microspectrofluorimetry. This system was sensitive ... More
Inositol 1,4,5-trisphosphate induces aggregation and release of 5-hydroxytryptamine from saponin-permeabilized human platelets.
AuthorsWatson SP, Ruggiero M, Abrahams SL, Lapetina EG
JournalJ Biol Chem
PubMed ID3082884
'Inositol 1,4,5-trisphosphate induces aggregation and the release of [3H]5-hydroxytryptamine from human platelets rendered permeable with saponin. This action of inositol 1,4,5-trisphosphate is associated with a significant formation of thromboxane B2, activation of phospholipase C, and phosphorylation of 20,000- and 40,000-dalton proteins, which are the substrates for myosin light chain kinase ... More
High-performance optical filters for fluorescence analysis.
AuthorsMarcus DA
JournalCell Motil Cytoskeleton
PubMed ID3180249
'Recent advances in thin film optical coating technology significantly improve the filters available for fluorescence spectroscopy. Bandpass and long- and shortpass filters with very sharply defined edges can provide from 10(-5) to 10(-6) blocking within 10-15 nm of the transmission region and are ideal for use as excitation and emission ... More
Microspectrofluorometry as a tool for investigation of non-calcium interactions of Indo-1.
AuthorsBancel F, Salmon JM, Vigo J, Viallet P
JournalCell Calcium
PubMed ID1540988
'Indo-1 is a fluorescent calcium probe used to measure intracellular free calcium concentrations. These measurements are often performed by comparing the fluorescence intensities of Indo-1-treated cells at two selected wavelengths corresponding to the maxima of the fluorescence spectra of the calcium-bound and calcium-free forms. In this study, we used an ... More
Three-photon induced fluorescence of the calcium probe Indo-1.
AuthorsSzmacinski H, Gryczynski I, Lakowicz JR
JournalBiophys J
PubMed ID8770232
'We report the calcium-dependent emission spectral properties of the calcium probe Indo-1 for three-photon excitation. We found that Indo-1 could be readily excited with the femtosecond pulses from a mode-locked Ti:sapphire laser at 885 nm. This wavelength is too long for two-photon excitation, which is expected to occur for wavelengths ... More
An oxidative stress-mediated death pathway in irradiated human leukemia cells mapped using multilaser flow cytometry.
AuthorsSheng-Tanner X, Bump EA, Hedley DW
JournalRadiat Res
PubMed ID9840183
'OCI/AML-2 acute myeloid leukemia cells were found to undergo apoptosis after treatment with y rays from a 137Cs source. Multilaser flow cytometry techniques using probes for live cell function were used to monitor the biochemical changes that occurred prior to the loss of surface membrane integrity. These showed increases in ... More
Intracellular calibration of the calcium indicator indo-1 in isolated fibers of Xenopus muscle.
AuthorsWesterblad H, Allen DG
JournalBiophys J
PubMed ID8842230
'Estimates of the free myoplasmic [Ca2+] ([Ca2+]i) with fluorescent dyes are complicated by the fact that some properties of these dyes are altered in the intracellular environment. In the present study indo-1 was used to measure [Ca2+]i in isolated muscle fibers from Xenopus frogs. Fluorescent ratio signals obtained from indo-1 ... More
Regulation of Ca2+ influx in myeloid cells. Role of plasma membrane potential, inositol phosphates, cytosolic free [Ca2+], and filling state of intracellular Ca2+ stores.
'To study the mediation of Ca2+ influx by second messengers in myeloid cells, we have combined the whole-cell patch clamp technique with microfluorimetric measurements of [Ca2+]i. Me2SO-differentiated HL-60 cells were loaded with the fluorescent Ca2+ indicator Indo-1, allowed to adhere to glass slides, and patch-clamped. Receptor agonists and Ca(2+)-ATPase inhibitors ... More
Functional characterization and potential applications for enhanced green fluorescent protein- and epitope-fused human M1 muscarinic receptors.
AuthorsWeill C, Galzi JL, Chasserot-Golaz S, Goeldner M, Ilien B
JournalJ Neurochem
PubMed ID10428077
'Four recombinant human M1 (hM1) muscarinic acetylcholine receptors (mAChRs) combining several modifications were designed and overexpressed in HEK293 cells. Three different fluorescent chimera were obtained through fusion of the receptor N terminus with enhanced green fluorescent protein (EGFP), potential glycosylation sites and a large part of the third intracellular (i3) ... More
Multiple spectral parameter imaging.
AuthorsWaggoner A, DeBiasio R, Conrad P, Bright GR, Ernst L, Ryan K, Nederlof M, Taylor D
JournalMethods Cell Biol
PubMed ID2648118
Quantitation of cytosolic [Ca2+] in whole perfused rat hearts using Indo-1 fluorometry.
AuthorsBrandes R, Figueredo VM, Camacho SA, Baker AJ, Weiner MW
JournalBiophys J
PubMed ID8298027
'Fluorometric determination of cytosolic calcium, [Ca2+]c, using Indo-1 in intact tissue, is limited by problems in obtaining calibration parameters for Indo-1 in vivo. Therefore, the goal of this study was to calibrate Indo-1 using in vitro constants, obtained from protein-containing reference solutions designed to produce similar Indo-1 spectral properties to ... More
The glucagon-induced activation of pyruvate dehydrogenase in hepatocytes is diminished by 4 beta-phorbol 12-myristate 13-acetate. A role for cytoplasmic Ca2+ in dehydrogenase regulation.
AuthorsStaddon JM, Hansford RG
JournalBiochem J
PubMed ID3593219
'Phenylephrine, vasopressin and glucagon each increased the amount of active (dephospho) pyruvate dehydrogenase (PDHa) in isolated rat hepatocytes. Treatment with 4 beta-phorbol 12-myristate 13-acetate (PMA) opposed the increase in PDHa caused by both phenylephrine and glucagon, but had no effect on the response to vasopressin: PMA alone had no effect ... More
Carbon monoxide inhibits depolarization-induced Ca rise and increases cyclic GMP in visceral smooth muscle cells.
AuthorsTrischmann U, Klöckner U, Isenberg G, Utz J, Ullrich V
JournalBiochem Pharmacol
PubMed ID1846546
'Monocytes were isolated from the urinary bladder of the guinea-pig. By means of the voltage clamp technique, whole cells were depolarized from -65 to +10 mV in order to increase the intracellular calcium concentration [Ca2+]i and to monitor this increase by means of the calcium activated potassium current IK.Ca. Superfusion ... More
Cytoplasmic free Ca2+ in Arabidopsis roots changes in response to touch but not gravity.
AuthorsLegué V, Blancaflor E, Wymer C, Perbal G, Fantin D, Gilroy S
JournalPlant Physiol
PubMed ID9232870
'Changes in cytoplasmic Ca2+ concentration ([Ca2+]i) have been proposed to be involved in signal transduction pathways in response to a number of stimuli, including gravity and touch. The current hypothesis proposes that the development of gravitropic bending is correlated with a redistribution of [Ca2+]i in gravistimulated roots. However, no study ... More
'Our understanding of the underlying mechanisms of Ca2+ signaling as well as our appreciation for its ubiquitous role in cellular processes has been rapidly advanced, in large part, due to the development of fluorescent Ca2+ indicators. In this chapter, we discuss some of the most common chemical Ca2+ indicators that ... More
The sarcolemmal mechanisms involved in the control of diastolic intracellular calcium in isolated rat cardiac trabeculae.
AuthorsLamont C, Eisner DA
JournalPflugers Arch
PubMed ID8781189
'We performed experiments using the calcium indicator Indo-1 to determine the relative roles of the sarcolemmal mechanisms involved in the regulation of diastolic intracellular calcium concentration ([Ca2+]i) in trabeculae from the rat heart. Ryanodine was used to eliminate sarcoplasmic reticulum (SR) function. In the functional absence of the SR, 76.8 ... More
Extracellular nucleotides elevate [Ca2+]i in rat osteoblastic cells by interaction with two receptor subtypes.
AuthorsReimer WJ, Dixon SJ
JournalAm J Physiol
PubMed ID1332491
'Extracellular nucleotides, through interaction with specific cell-surface receptors, mediate a variety of biological responses, including elevation of cytosolic free Ca2+ concentration ([Ca2+]i) in a number of cell types. The effects of nucleotides on [Ca2+]i in the rat osteoblastic cell line UMR-106 were studied by fluorescence spectrophotometry of indo-1-loaded cells. In ... More
Measurement of free Zn2+ ion concentration with the fluorescent probe mag-fura-2 (furaptra).
AuthorsSimons TJ
JournalJ Biochem Biophys Methods
PubMed ID8409208
'The fluorescent probe mag-fura-2, previously used to measure [Mg2+], can also be used to measure [Zn2+]. The peak in the excitation spectrum occurs at 323 nm for Zn2+, compared with 335 nm for Ca2+ and Mg2+. This allows simultaneous measurements of [Zn2+] and either [Ca2+] or [Mg2+], by using 3 ... More
The calcium feedback signal in the phototransduction cascade of vertebrate rods.
AuthorsGray-Keller MP, Detwiler PB
JournalNeuron
PubMed ID7524559
'Intracellular free Ca (Cai) was measured in functionally intact rod outer segments in darkness and during light responses using the fluorescent Ca indicator Indo-dextran. In darkness, Cai was 554 +/- 25 nM (n = 28) for -85 +/- 2 pA of circulating dark current (Id) and declined in saturating light ... More
Regulated Ca2+ signalling through leukocyte CD11b/CD18 integrin.
AuthorsAltieri DC, Stamnes SJ, Gahmberg CG
JournalBiochem J
PubMed ID1361124
'General mechanisms of adhesion in the immune response are coordinated by the leukocyte integrins CD11/CD18. The possible participation of these differentiation molecules in early events of transmembrane signalling was investigated. Monoclonal antibody (mAb) cross-linking of CD18, the integrin beta subunit ubiquitously expressed by all leukocytes, increased the cytosolic free Ca2+ ... More
Sodium-calcium exchange.
Authors
JournalScience
PubMed ID1848371
Methods for calibration of fluorescent calcium indicators in skeletal muscle fibers.
AuthorsWesterblad H, Allen DG
JournalBiophys J
PubMed ID8011925
Fura-2: a powerful new tool for measuring and imaging [Ca2+]i in single cells.
AuthorsPoenie M, Tsien R
JournalProg Clin Biol Res
PubMed ID3754335
Flow cytometric analysis of intracellular calcium mobilization.
AuthorsRansom JT, DiGiusto DL, Cambier J
JournalMethods Enzymol
PubMed ID3496517
Fluorescence measurement and photochemical manipulation of cytosolic free calcium.
AuthorsTsien RY
JournalTrends Neurosci
PubMed ID2469158
Fluorescent probes of cell signaling.
AuthorsTsien RY
JournalAnnu Rev Neurosci
PubMed ID2648950
Calcium transients during mitosis: observations in flux.
AuthorsHepler PK
JournalJ Cell Biol
PubMed ID2687283
Rapid simultaneous estimation of intracellular calcium and pH.
Determination of intracellular calcium in vital cells by flow-cytometry.
AuthorsValet G, Raffael A, Rüssmann L
JournalNaturwissenschaften
PubMed ID4079984
Intracellular ionized calcium.
AuthorsJune CH, Rabinovitch PS
JournalMethods Cell Biol
PubMed ID7861962
Practical aspects of measuring [Ca2+] with fluorescent indicators.
AuthorsKao JP
JournalMethods Cell Biol
PubMed ID8201975
Excitation-induced Ca2+ dynamics in sympathetic neurons measured with conventional epifluorescence and confocal UV laser-scanning microscopes.
AuthorsHua SY, Kuba K, Nohmi M
JournalJpn J Physiol
PubMed ID8271488
A depolarization-induced rise in intracellular Ca2+ in cultured bullfrog sympathetic ganglion cells depended non-linearly on Ca2+ influx and membrane depolarization, spread inwardly faster than that without Ca(2+)-induced Ca2+ release (CICR), and was blocked by dantrolene and ryanodine. Ca2+ entry and a graded activation of CICR thus induce the "Ca2+ transient." ... More
Microinjection of a conserved peptide sequence of p34cdc2 induces a Ca2+ transient in oocytes.
AuthorsPicard A, Cavadore JC, Lory P, Bernengo JC, Ojeda C, Dorée M
JournalScience
PubMed ID2153316
The product of the yeast cell cycle control gene cdc2, and its homologs in higher eukaryotes (p34cdc2), all contain a perfectly conserved sequence of 16 amino acids that has not been found in any other protein sequence. Microinjection of this peptide triggers a specific increase in the concentration of intracellular ... More
The kinetics of calcium binding to fura-2 and indo-1.
AuthorsJackson AP, Timmerman MP, Bagshaw CR, Ashley CC
JournalFEBS Lett
PubMed ID3108033
The kinetics of Ca2+ dissociation from fura-2 and indo-1 were measured using a stopped-flow spectrofluorimeter. The dissociation rate constants were 84 s-1 and 130 s-1, respectively, in 0.1 M KC1 at 20 degrees C. The rate constants were insensitive to pH over the range 7.0 to 8.0. The second order ... More
Amyloid beta-peptide(25-35) changes [Ca2+] in hippocampal neurons.
AuthorsMogensen HS, Beatty DM, Morris SJ, Jorgensen OS
JournalNeuroreport
PubMed ID9631466
Insoluble aggregates of the amyloid beta-peptide (A beta) is a major constituent of senile plaques found in brains of Alzheimer disease (AD) patients. The detrimental effects of aggregated A beta is associated with an increased intracellular Ca2+ concentration ([Ca2+]i). We examined the effects of A beta(25-35) on [Ca2+]i and intracellular ... More
The effects of pH and temperature on fluorescent calcium indicators as determined with Chelex-100 and EDTA buffer systems.
AuthorsLattanzio FA
JournalBiochem Biophys Res Commun
PubMed ID2118341
A novel method of determining the apparent dissociation constants of fluorescent calcium indicators is described which utilizes Chelex-100 ion exchange resin and 45Ca. The affinity for calcium of indicators fluo-3, fura-2 and indo-1 measured at either 22 degrees or 37 degrees C decreases as pH is decreased from 7.4 to ... More
Continuous monitoring of transport by fluorescence on cells and vesicles.
AuthorsEidelman O, Cabantchik ZI
JournalBiochim Biophys Acta
PubMed ID2686755
Fluorescence techniques are gaining wider applicability in the field of membrane transport due to their high temporal resolution, modest demand for biological material and the kinetic information which is made available by fluorescence tracings. The development of novel fluorescent substrates for particular transport systems and of novel fluorescent indicators for ... 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
Molecular excitation by the simultaneous absorption of two photons provides intrinsic three-dimensional resolution in laser scanning fluorescence microscopy. The excitation of fluorophores having single-photon absorption in the ultraviolet with a stream of strongly focused subpicosecond pulses of red laser light has made possible fluorescence images of living cells and other ... 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
Calcium mobilization by dual receptors during fertilization of sea urchin eggs.
AuthorsLee HC, Aarhus R, Walseth TF
JournalScience
PubMed ID8392749
Fertilization is accompanied by a transient increase in the concentration of intracellular Ca2+, which serves as a signal for initiating development. Some of the Ca2+ appears to be released from intracellular stores by the binding of inositol trisphosphate (IP3) to its receptor. However, in sea urchin eggs, other mechanisms appear ... More
Dye loading with patch pipettes.
AuthorsEilers J, Konnerth A,
JournalCSH Protoc
PubMed ID20147143
This protocol describes the loading of individual cells with fluorescent probes via patch pipettes. The patch-clamp methodology has been successfully used for single-cell dye labeling in cultured neurons, brain slices, and in vivo preparations. A broad range of dyes can be used with this loading technique. Markers for morphological reconstruction ... More
Calcium influx via TRP channels is required to maintain PIP2 levels in Drosophila photoreceptors.
AuthorsHardie RC, Raghu P, Moore S, Juusola M, Baines RA, Sweeney ST
JournalNeuron
PubMed ID11343651
The trp (transient receptor potential) gene encodes a Ca2+ channel responsible for the major component of the phospholipase C (PLC) mediated light response in Drosophila. In trp mutants, maintained light leads to response decay and temporary total loss of sensitivity (inactivation). Using genetically targeted PIP2-sensitive inward rectifier channels (Kir2.1) as ... More
Cross-linking of surface immunoglobulin on B lymphocytes induces both intracellular Ca2+ release and Ca2+ influx: analysis with indo-1.
AuthorsBijsterbosch MK, Rigley KP, Klaus GG
JournalBiochem Biophys Res Commun
PubMed ID3487320
The new Ca2+-probe indo-1 has a high fluorescence intensity, which allows low intracellular dye loadings. Stimulation of indo-1-loaded mouse B cells with anti-Ig antibodies provoked rapid rise of free cytoplasmic Ca2+ from 100 nM to greater than 1 microM, followed by a decline to a plateau at 300-400 nM. The ... More
Rhythmic exocytosis stimulated by GnRH-induced calcium oscillations in rat gonadotropes.
AuthorsTse A, Tse FW, Almers W, Hille B
JournalScience
PubMed ID8385366
In pituitary gonadotropes, gonadotropin-releasing hormone (GnRH) induces the rhythmic release of Ca2+ from an inositol 1,4,5-trisphosphate (IP3)-sensitive store. Simultaneous measurement of the concentration of cytosolic free Ca2+ ([Ca2+]i) and exocytosis in single identified gonadotropes showed that each elevation of [Ca2+]i induced a burst of exocytosis. These phenomena were largely suppressed ... More
Calcium sequestration by isolated sarcoplasmic reticulum: real-time monitoring using ratiometric dual-emission spectrofluorometry and the fluorescent calcium-binding dye indo-1.
AuthorsO'Brien PJ
JournalMol Cell Biochem
PubMed ID2374546
This study demonstrates a simple, rapid, and reproducible microassay for real-time monitoring of Ca2(+)-sequestration by isolated sarcoplasmic reticulum (SR) using ratiometric dual-emission spectrofluorometry and the fluorescent calcium-binding dye indo-1. The SR membranes were isolated by differential centrifugation and suspended in a medium including Ca2+, indo-1, ATP and oxalate. As Ca2+ ... More
Electrophysiological responses to bradykinin and microinjected inositol polyphosphates in neuroblastoma cells. Possible role of inositol 1,3,4-trisphosphate in altering membrane potential.
Addition of bradykinin to mouse N1E-115 neuroblastoma cells evokes a rapid but transient rise in cytoplasmic free Ca2+ concentration ([Ca2+]i). The [Ca2+]i rise is accompanied by a transient membrane hyperpolarization, due to a several-fold increase in K+ conductance, followed by a prolonged depolarizing phase. Pretreatment of the cells with a ... More
Calcium-dependent fluorescence lifetimes of Indo-1 for one- and two-photon excitation of fluorescence.
AuthorsSzmacinski H, Gryczynski I, Lakowicz JR
JournalPhotochem Photobiol
PubMed ID8234466
We characterized the fluorescence intensity decays of Indo-1, which is commonly used as an emission wavelength-ratiometric calcium probe. The apparent lifetime of the long-wavelength side of the emission of Indo-1 is dependent on Ca2+. This long-wavelength emission displays the characteristics of an excited-state reaction, that is, a negative preexponential component ... 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
Does the use of DM-nitrophen, nitr-5, or diazo-2 interfere with the measurement of indo-1 fluorescence?
AuthorsHadley RW, Kirby MS, Lederer WJ, Kao JP
JournalBiophys J
PubMed ID8312490
Emission spectra of the photolabile Ca2+ chelators DM-nitrophen, nitr-5, and diazo-2 were studied alone, and in the presence of indo-1, to investigate potential interactions that would make the simultaneous manipulation and ratiometric measurement of the intracellular Ca2+ concentration difficult. Neither diazo-2 nor its photoproduct were found to be significantly fluorescent, ... More
Optimum spectral windows to minimize quantum noise of ratiometric intracellular fluorescent probes.
AuthorsStern MD, Spurgeon HA, Hansford R, Lakatta EG, Capogrossi MC
JournalCell Calcium
PubMed ID2620339
When fluorescent indicators are used to measure intracellular ligands in single cells, the quality of the data is usually limited by quantum (shot) noise. For indicators which shift excitation or emission wavelengths upon ligand binding, a ratiometric method is usually employed. In choosing the spectral windows for excitation or collection ... More
Activation of G-proteins induces Ca2+ oscillations with hyperpolarizing K+ currents in pancreatic beta-cells.
AuthorsLund PE, Hellman B
JournalSecond Messengers Phosphoproteins
PubMed ID1345342
Activation of G-proteins by internal perfusion with GTP-gamma-S or external application of carbachol resulted in oscillations of cytoplasmic Ca2+ in isolated mouse pancreatic beta-cells. The Ca2+ transients were associated with the generation of K+ currents sufficiently pronounced to induce marked pulses of hyperpolarization. The oscillatory G-protein response remained largely unaffected ... More
Adapting a compact confocal microscope system to a two-photon excitation fluorescence imaging architecture.
AuthorsDiaspro A, Corosu M, Ramoino P, Robello M
JournalMicrosc Res Tech
PubMed ID10544334
Within the framework of a national National Institute of Physics of Matter (INFM) project, we have realised a two-photon excitation (TPE) fluorescence microscope based on a new generation commercial confocal scanning head. The core of the architecture is a mode-locked Ti:Sapphire laser (Tsunami 3960, Spectra Physics Inc., Mountain View, CA) ... More
Sodium current-induced release of calcium from cardiac sarcoplasmic reticulum.
AuthorsLeblanc N, Hume JR
JournalScience
PubMed ID2158146
The role of sodium-calcium exchange at the sarcolemma in the release of calcium from cardiac sarcoplasmic reticulum was investigated in voltage-clamped, isolated cardiac myocytes. In the absence of calcium entry through voltage-dependent calcium channels, membrane depolarization elicited release of calcium from ryanodine-sensitive internal stores. This process was dependent on sodium ... More
Parallel changes in nuclear and cytosolic calcium in mouse pancreatic beta-cells.
AuthorsBrown GR, Köhler M, Berggren PO
JournalBiochem J
PubMed ID9271099
In the neuroendocrine pancreatic beta-cell, elevations in intracellular Ca2+ lead to insulin secretion and the initiation of gene transcription. However, the relationship between cytosolic and nuclear Ca2+ in these cells is unknown. The Ca2+ permeability of the nuclear membrane would therefore determine if Ca2+ could play a direct role in ... More
Ion channel activity and tip growth: tip-localized stretch-activated channels generate an essential Ca2+ gradient in the oomycete Saprolegnia ferax.
AuthorsGarrill A, Jackson SL, Lew RR, Heath IB
JournalEur J Cell Biol
PubMed ID7687216
The plasma membrane of tip-growing hyphae of the oomycete Saprolegnia ferax contains stretch-activated (SA) Ca(2+)-permeable and Ca(2+)-activated K+ ion channels. Patch clamp measurements on protoplasts derived from specific regions of hyphae demonstrated that SA channels were most abundant in the tip. Gadolinium (Gd3+) inhibited SA channel activity and stopped tip ... More
The use of fura-2 for estimating Ca buffers and Ca fluxes.
AuthorsNeher E
JournalNeuropharmacology
PubMed ID8606791
The compound fura-2 (Grynkiewicz et al., J. Biol. Chem. 260, 3440-3450, 1985) is generally known as an indicator dye for measuring the concentration of free calcium ([Ca2+]) inside living cells. It should be appreciated, however, that this is not what it actually is. More accurately, it is a divalent metal ... More
Modulation of the intracellular Ca2+ and inositol trisphosphate concentrations in murine T lymphocytes by the glycosylphosphatidylinositol-anchored protein sgp-60.
AuthorsMaschek BJ, Zhang W, Rosoff PM, Reiser H
JournalJ Immunol
PubMed ID8468465
Stimulation of T lymphocytes via the TCR/CD3 complex initiates a series of intracellular biochemical events. Among the earliest cellular responses are the stimulation of the turnover of phosphatidylinositol and an increase in the intracellular Ca2+ concentration. The significance of these second messengers for distal cellular responses such as lymphokine production ... More
Gibberellic acid and abscisic acid coordinately regulate cytoplasmic calcium and secretory activity in barley aleurone protoplasts.
AuthorsGilroy S, Jones RL
JournalProc Natl Acad Sci U S A
PubMed ID1533046
The effects of gibberellic acid (GA3) and abscisic acid (ABA) on the temporal and spatial dynamics of cytosolic calcium concentration ([Ca2+]i) in aleurone protoplasts of barley (Hordeum vulgare L. cv. Himalaya) were measured by using fluorescence ratio analysis and confocal microscopy. After 4-6 h of treatment, GA3 induced a sustained ... More
Two-photon thermal bleaching of single fluorescent molecules.
AuthorsChirico G, Cannone F, Baldini G, Diaspro A
JournalBiophys J
PubMed ID12524312
We have studied the fluorescence emission by two-photon excitation of four dyes widely used for bioimaging studies, rhodamine 6G, fluorescein, pyrene and indo-1 at the single molecule level. The single dye molecules, spread on a glass substrate by spin coating, show a constant fluorescence output until a sudden transition to ... More
Interactions between inositol trisphosphate receptors and fluorescent Ca2+ indicators.
AuthorsMorris SA, Correa V, Cardy TJ, O'Beirne G, Taylor CW
JournalCell Calcium
PubMed ID10326680
Fura-2 and BAPTA were previously shown to be competitive antagonists of inositol trisphosphate (InsP3) receptors, but for practical reasons the analyses were performed at pH 8.3. We recently developed a scintillation proximity assay (SPA) for pure cerebellar InsP3 receptors which allows low affinity interactions to be characterized and is readily ... More
Attachment of A172 human glioblastoma cells affects calcium signalling: a comparison of image cytometry, flow cytometry, and spectrofluorometry.
AuthorsSzöllösi J, Feuerstein BG, Hyun WC, Das MK, Marton LJ
JournalCytometry
PubMed ID1794252
The intracellular free calcium concentration ([Ca2+]i) of indo-1 loaded A172 human glioblastoma cells stimulated by platelet-derived growth factor (PDGF) was studied in cell suspensions by flow cytometry and spectrofluorometry and in confluent monolayers by laser image cytometry and spectrofluorometry. With all three techniques, the percentage of responsive cells, peak [Ca2+]i, ... More
Phorbol ester (12-O-tetradecanoylphorbol 13-acetate) partially inhibits rapid intracellular free calcium transients triggered by anti-immunoglobulin in murine lymphocytes.
AuthorsOwen CS
JournalJ Biol Chem
PubMed ID3257754
The highly fluorescent probes Indo-1 and Fura-2 were employed to detect intracellular calcium responses in murine splenic lymphocytes following cross-linking of cell surface Ig. Inhibition by phorbol ester (12-O-tetradecanoylphorbol 13-acetate) was rapid and showed a strong preference for the very transient phase of the response which has been identified as ... More