Detection of plasma membrane Ca(2+)-ATPase activity in mouse T lymphocytes by flow cytometry using fluo-3-loaded vesicles.
AuthorsTelford WG, Miller RA
JournalCytometry
PubMed ID8800557
The plasma membrane Ca(2+)-ATPase (PMCA) is the primary means by which many cell types pump calcium out of the cytosol following release of calcium from internal stores, returning intracellular calcium concentrations to normal levels. Traditional methods for measuring PMCA activity utilizing isotopic calcium uptake into inside-out (IO) membrane vesicles have ... More
Extracellular Mg2+ induces an intracellular Ca2+ wave during oocyte activation in the marine shrimp Sicyonia ingentis.
AuthorsLindsay LL, Hertzler PL, Clark WH
JournalDev Biol
PubMed ID1628759
'In contrast to most systems in which oocyte activation is triggered by the fertilizing sperm, Sicyonia ingentis oocytes are activated by seawater Mg2+ during spawning. S. ingentis oocytes were spawned into Mg(2+)-free seawater and microinjected with the fluorescent Ca2+ indicator Fluo-3 to study the effects of added Mg2+ on intracellular ... More
Caged nicotinic acid adenine dinucleotide phosphate. Synthesis and use.
AuthorsLee HC, Aarhus R, Gee KR, Kestner T
JournalJ Biol Chem
PubMed ID9020130
'Nicotinic acid adenine dinucleotide phosphate (NAADP) is a metabolite of NADP with Ca2+ mobilizing activity. The Ca2+ release mechanism activated by NAADP as well as the Ca2+ stores that it acts on are different from those activated by either cyclic ADP-ribose or inositol 1,4,5-trisphosphate (IP3) (Lee, H. C., and Aarhus, ... More
Mechanisms of subcellular cytosolic Ca2+ signaling evoked by stimulation of the vasopressin V1a receptor.
'Receptor activation may result in distinct subcellular patterns of Ca2+ release. To define the subcellular distribution of Ca2+i signals induced by stimulation of the vasopressin V1a receptor, we expressed the cloned receptor in Xenopus oocytes. Oocytes were then loaded with fluo-3 and observed using confocal microscopy. Vasopressin induced a single ... More
Patch-clamp detection of macromolecular translocation along nuclear pores.
AuthorsBustamante JO, Varanda WA
JournalBraz J Med Biol Res
PubMed ID9698781
'The present paper reviews the application of patch-clamp principles to the detection and measurement of macromolecular translocation along the nuclear pores. We demonstrate that the tight-seal ''gigaseal'' between the pipette tip and the nuclear membrane is possible in the presence of fully operational nuclear pores. We show that the ability ... More
Core mutations that promote the calcium-induced allosteric transition of bovine recoverin.
AuthorsBaldwin AN, Ames JB
JournalBiochemistry
PubMed ID9860856
'Recoverin is a small calcium binding protein involved in regulation of the phototransduction cascade in retinal rod cells. It functions as a calcium sensor by undergoing a cooperative, ligand-dependent conformational change, resulting in the extrusion of the N-terminal myristoyl group from a hydrophobic pocket. To test the role of certain ... More
Synaptic-type acetylcholine receptors raise intracellular calcium levels in neurons by two mechanisms.
AuthorsRathouz MM, Berg DK
JournalJ Neurosci
PubMed ID7965089
'Nicotinic acetylcholine receptors (AChRs) located in the postsynaptic membrane on neurons are responsible for mediating fast, excitatory synaptic transmission. If synaptic AChRs are also highly permeable to calcium as reported recently for several kinds of neuronal AChRs, the synaptic receptors could regulate calcium-dependent events in the neurons in concert with ... 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
Coexpression of Drosophila TRP and TRP-like proteins in Xenopus oocytes reconstitutes capacitative Ca2+ entry.
AuthorsGillo B, Chorna I, Cohen H, Cook B, Manistersky I, Chorev M, Arnon A, Pollock JA, Selinger Z, Minke B
JournalProc Natl Acad Sci U S A
PubMed ID8943075
'Capacitative Ca2+ entry is a component of the inositol-lipid signaling in which depletion of inositol 1,4,5-trisphosphate (InsP3)-sensitive Ca2+ stores activates Ca2+ influx by a mechanism that is still unknown. This pathway plays a central role in cellular signaling, which is mediated by many hormones, neurotransmitters, and growth factors. Studies of ... More
Dopamine modulates GABAc receptors mediating inhibition of calcium entry into and transmitter release from bipolar cell terminals in tiger salamander retina.
AuthorsWellis DP, Werblin FS
JournalJ Neurosci
PubMed ID7623108
'Using optical recording techniques, we directly monitored pre- and postsynaptic calcium dynamics at bipolar cell terminals while inhibiting synaptic release with applied GABA and modulating inhibition with dopamine. To monitor pre-synaptic activity, individual bipolar cells in the retinal slice were filled with either fura-2 or fluo-3 through a patch electrode. ... More
Ca2+ regulation of gelsolin by its C-terminal tail.
AuthorsLin KM, Mejillano M, Yin HL
JournalJ Biol Chem
PubMed ID10862770
'Gelsolin is activated by Ca(2+) to sever actin filaments. Ca(2+) regulation is conferred on the N-terminal half by the C-terminal half. This paper seeks to understand how Ca(2+) regulates gelsolin by testing the "tail helix latch hypothesis," which is based on the structural data showing that gelsolin has a C-terminal ... More
Hydrogen peroxide attenuates store-operated calcium entry and enhances calcium extrusion in thyroid FRTL-5 cells.
AuthorsTörnquist K, Vainio PJ, Björklund S, Titievsky A, Dugué B, Tuominen RK
JournalBiochem J
PubMed ID10998346
'Redox modulation participates in the regulation of intracellular free calcium concentration ([Ca(2+)](i)) in several cell types. In thyroid cells, including FRTL-5 cells, changes in [Ca(2+)](i) regulate several important functions, including the production of H(2)O(2) (hydrogen peroxide). As H(2)O(2) is of crucial importance for the production of thyroid hormones, we investigated ... More
Imaging of calcium transients in skeletal muscle fibers.
AuthorsVergara J, DiFranco M, Compagnon D, Suarez-Isla BA
JournalBiophys J
PubMed ID2015378
'Epifluorescence images of Ca2+ transients elicited by electrical stimulation of single skeletal muscle fibers were studied with fast imaging techniques that take advantage of the large fluorescence signals emitted at relatively long wavelengths by the dyes fluo-3 and rhod-2 in response to binding of Ca2+ ions, and of the suitable ... More
The control of calcium release in heart muscle.
AuthorsCannell MB, Cheng H, Lederer WJ
JournalScience
PubMed ID7754384
'The control of calcium release from intracellular stores (the sarcoplasmic reticulum) in cardiac muscle was examined with the use of a confocal microscope and voltage clamp techniques. Depolarization evoked graded calcium release by altering the extent of spatial and temporal summation of elementary calcium release events called "calcium sparks." These ... More
Localization in the II-III loop of the dihydropyridine receptor of a sequence critical for excitation-contraction coupling.
AuthorsNakai J, Tanabe T, Konno T, Adams B, Beam KG
JournalJ Biol Chem
PubMed ID9737952
'Skeletal and cardiac muscles express distinct isoforms of the dihydropyridine receptor (DHPR), a type of voltage-gated Ca2+ channel that is important for excitation-contraction (EC) coupling. However, entry of Ca2+ through the channel is not required for skeletal muscle-type EC coupling. Previous work (Tanabe, T., Beam, K. G., Adams, B. A., ... More
Chemical depolarization-induced SR calcium release in triads isolated from rabbit skeletal muscle.
AuthorsIkemoto N, Yano M, el-Hayek R, Antoniu B, Morii M
JournalBiochemistry
PubMed ID8086413
'Excitation-Ca2+ release coupling properties in the heavy microsomal fraction of the rabbit skeletal muscle enriched in triads were investigated by following the same type of approach used for the studies of excitation-contraction coupling in the skinned fiber system. Incubation of the triads with Mg-ATP in a solution containing 150 mM ... More
A comparison of the effects of ATP and tetracaine on spontaneous Ca(2+) release from rat permeabilised cardiac myocytes.
AuthorsSmith GL, O'Neill SC
JournalJ Physiol
PubMed ID11432990
'1. Fluo-3 fluorescence measurements were made in isolated beta-escin permeablised rat cardiac myocytes using confocal microscopy. Perfusion of a mock intracellular solution containing 0.22-0.23 microM Ca(2+) and 5 mM ATP elicited regular waves of Ca(2+) (approximately every 5 s) due to spontaneous release of Ca(2+) from the sarcoplasmic reticulum (SR). ... More
Determining zinc with commonly used calcium and zinc fluorescent indicators, a question on calcium signals.
AuthorsMartin JL, Stork CJ, Li YV
JournalCell Calcium
PubMed ID16764924
'Investigations into the roles of Ca(2+) and Zn(2+) in cell biology have been facilitated by the development of sensitive fluorometric probes that have enabled the measurement of Ca(2+) or Zn(2+) in both extracellular and intracellular environments. It is critical to be aware of the specificity and relative selectivity of a ... More
cGMP mobilizes intracellular Ca2+ in sea urchin eggs by stimulating cyclic ADP-ribose synthesis.
AuthorsGalione A, White A, Willmott N, Turner M, Potter BV, Watson SP
JournalNature
PubMed ID7692303
'Many hormones or neurotransmitters act at cell surface receptors to increase the intracellular free calcium concentration, triggering a wide range of cellular responses. As the source of this Ca2+ is often internal stores, additional messengers are required to convey the hormonal message from the plasma membrane. Cyclic ADP-ribose (cADPR) has ... More
Type III InsP3 receptor channel stays open in the presence of increased calcium.
AuthorsHagar RE, Burgstahler AD, Nathanson MH, Ehrlich BE
JournalNature
PubMed ID9817204
'The inositol 1,4,5-trisphosphate receptor (InsP3R) is the main calcium(Ca2+) release channel in most tissues. Three isoforms have been identified, but only types I and II InsP3R have been characterized. Here we examine the functional properties of the type III InsP3R because this receptor is restricted to the trigger zone from ... More
A preferred amplitude of calcium sparks in skeletal muscle.
AuthorsRíos E, Shirokova N, Kirsch WG, Pizarro G, Stern MD, Cheng H, González A
JournalBiophys J
PubMed ID11159393
'In skeletal and cardiac muscle, calcium release from the sarcoplasmic reticulum, leading to contraction, often results in calcium sparks. Because sparks are recorded by confocal microscopy in line-scanning mode, their measured amplitude depends on their true amplitude and the position of the spark relative to the scanned line. We present ... More
Calcium-induced inactivation of NMDA receptor-channels evolves independently of run-down in cultured rat brain neurones.
AuthorsMedina I, Filippova N, Bakhramov A, Bregestovski P
JournalJ Physiol
PubMed ID8887753
'1. Calcium-induced transient inactivation of NMDA receptor (NMDAR) channels was studied in cultured rat hippocampal and cerebellar granule neurones using patch-clamp techniques and confocal scanning microscopy. 2. During whole-cell recordings, in the presence of 2 mM external Ca2+, conditioning (2-20 s) pulses of NMDA (20-100 microM) caused a transient decrease ... More
Carbachol induces oscillations in membrane potential and intracellular calcium in a colonic tumor cell line, HT-29.
AuthorsSand P, Svenberg T, Rydqvist B
JournalAm J Physiol
PubMed ID9357762
'The patch-clamp technique was used to study the effects of carbachol (CCh) on HT-29 cells. During CCh exposure, the cells (n = 23) depolarized close to the equilibrium potential for Cl- (E(Cl-); -48 mV) and the membrane potential then started to oscillate (16/23 cells). In voltage-clamp experiments, similar oscillations in ... More
Functional heterogeneity of calcium release by inositol trisphosphate in single Purkinje neurones, cultured cerebellar astrocytes, and peripheral tissues.
AuthorsKhodakhah K, Ogden D
JournalProc Natl Acad Sci U S A
PubMed ID8506344
'Purkinje neurones of the cerebellar cortex are rich in receptors for the Ca-mobilizing second messenger inositol trisphosphate (InsP3) in association with intracellular Ca stores. Cytosolic Ca ions are important in regulating neuronal excitability but it has proved difficult to demonstrate InsP3-evoked release of Ca in mammalian central neurones directly. Intracellular ... More
Intercellular Ca2+ waves in rat heart muscle.
AuthorsLamont C, Luther PW, Balke CW, Wier WG
JournalJ Physiol
PubMed ID9769412
'1. Confocal laser scanning microscopy was used to visualize intercellular transmission of Ca2+ waves in intact rat ventricular trabeculae micro-injected with the calcium indicator fluo-3. 2. Ca2+ waves usually failed to be transmitted from cell to cell. At identified individual end-to-end cell contacts, successful transmission interspersed with failure, which sometimes ... More
Thrombin-induced phosphorylation and activation of Ca(2+)-sensitive cytosolic phospholipase A2 in human platelets.
AuthorsKramer RM, Roberts EF, Manetta JV, Hyslop PA, Jakubowski JA
JournalJ Biol Chem
PubMed ID8253817
'Receptor-mediated activation of human platelets by thrombin initiates a series of rapid biochemical events that include activation of phospholipase A2 to liberate arachidonic acid for further conversion to thromboxane A2. The identity of the phospholipase A2 involved has not been clear. Here we show by immunochemical analysis that human platelets ... More
Cyclic ADP-ribose in insulin secretion from pancreatic beta cells.
AuthorsTakasawa S, Nata K, Yonekura H, Okamoto H
JournalScience
PubMed ID8420005
'Inositol 1,4,5-trisphosphate (IP3) is thought to be a second messenger for intracellular calcium mobilization. However, in a cell-free system of islet microsomes, cyclic adenosine diphosphate-ribose (cADP-ribose), a nicotinamide adenine dinucleotide (NAD+) metabolite, but not IP3, induced calcium release. In digitonin-permeabilized islets, cADP-ribose and calcium, but not IP3, induced insulin secretion. ... More
Nitric oxide-induced mobilization of intracellular calcium via the cyclic ADP-ribose signaling pathway.
AuthorsWillmott N, Sethi JK, Walseth TF, Lee HC, White AM, Galione A
JournalJ Biol Chem
PubMed ID8631983
'Cyclic adenosine diphosphate ribose (cADPR) is a potent endogenous calcium-mobilizing agent synthesized from beta-NAD+ by ADP-ribosyl cyclases in sea urchin eggs and in several mammalian cells (Galione, A., and White, A. (1994) Trends Cell Biol. 4, 431 436). Pharmacological studies suggest that cADPR is an endogenous modulator of Ca2+-induced Ca2+ ... More
Dendritic calcium transients in the leech giant glial cell in situ.
AuthorsLohr C, Deitmer JW
JournalGlia
PubMed ID10384876
'Glial cells have been shown to respond to neuronal activity with changes in the membrane potential and the intracellular Ca2+ concentration. In order to get closer to glial structures associated with neuronal synapses, we have now looked at Ca2+ signalling in the glial processes ("glial dendrites") in response to neurotransmitters ... More
Agonist-specific behaviour of the intracellular Ca2+ response in rat hepatocytes.
AuthorsChatton JY, Cao Y, Stucki JW
JournalBiochem J
PubMed ID9371717
'A variety of agonists stimulate in hepatocytes a response that takes the shape of repetitive cytosolic free Ca2+ transients called Ca2+ oscillations. The shape of spikes and the pattern of oscillations in a given cell differ depending on the agonist of the phosphoinositide pathway that is applied. In this study, ... More
The path of calcium in cytosolic calcium oscillations: a unifying hypothesis.
AuthorsJaffe LF
JournalProc Natl Acad Sci U S A
PubMed ID1946414
'Data from 42 systems have been assembled in which the overall spatial course of relatively natural, intracellular calcium pulses has been or can be determined. These include 21 cases of solitary pulses in activating eggs and 21 cases of periodic (as well as solitary) pulses in various fully active cells. ... More
Polarized expression of Ca2+ pumps in pancreatic and salivary gland cells. Role in initiation and propagation of [Ca2+]i waves.
AuthorsLee MG, Xu X, Zeng W, Diaz J, Kuo TH, Wuytack F, Racymaekers L, Muallem S
JournalJ Biol Chem
PubMed ID9188473
'The present study was aimed at localization of plasma membrane (PMCA) and intracellular (SERCA) Ca2+ pumps and characterizing their role in initiation and propagation of Ca2+ waves. Specific and polarized expression of Ca2+ pumps was observed in all epithelial cells examined. Immunolocalization revealed expression of PMCA in both the basolateral ... More
Calmodulin-dependent regulation of inducible and neuronal nitric-oxide synthase.
AuthorsLee SJ, Stull JT
JournalJ Biol Chem
PubMed ID9765272
'Neuronal and endothelial nitric-oxide synthases depend upon Ca2+/calmodulin for activation, whereas the activity of the inducible nitric-oxide synthase is Ca2+-independent, presumably due to tightly bound calmodulin. To study these different mechanisms, a series of chimeras derived from neuronal and inducible nitric- oxide synthases were analyzed. Chimeras containing only the oxygenase ... More
Real-time imaging of neurons retrogradely and anterogradely labelled with calcium-sensitive dyes.
AuthorsO'Donovan MJ, Ho S, Sholomenko G, Yee W
JournalJ Neurosci Methods
PubMed ID8474261
'Membrane-impermeant calcium indicator dyes were used to retrogradely label dorsal root ganglia, spinal motoneurons and interneurons in the spinal cord of the chick embryo. The dyes were also used to label anterogradely primary afferent axons in the spinal cord and synaptic endings in the ciliary ganglion. Labelled neurons were imaged ... More
A primitive ATP receptor from the little skate Raja erinacea.
AuthorsDranoff JA, O'Neill AF, Franco AM, Cai SY, Connolly GC, Ballatori N, Boyer JL, Nathanson MH
JournalJ Biol Chem
PubMed ID10900200
'P2Y ATP receptors are widely expressed in mammalian tissues and regulate a broad range of activities. Multiple subtypes of P2Y receptors have been identified and are distinguished both on a molecular basis and by pharmacologic substrate preference. Functional evidence suggests that hepatocytes from the little skate Raja erinacea express a ... More
Fluorescent indicators for cytosolic calcium based on rhodamine and fluorescein chromophores.
AuthorsMinta A, Kao JP, Tsien RY
JournalJ Biol Chem
PubMed ID2498308
'A new group of fluorescent indicators with visible excitation and emission wavelengths has been synthesized for measurements of cytosolic free Ca2+. The five compounds, "rhod-1," "rhod-2," "fluo-1," "fluo-2," and "fluo-3" (Figs. 2 and 3), combine the 8-coordinate tetracarboxylate chelating site of 1,2-bis(2-amino-phenoxyethane-N,N,N'',N''-tetraacetic acid with a xanthene chromophore to give a ... 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
Electric field-directed fibroblast locomotion involves cell surface molecular reorganization and is calcium independent.
AuthorsBrown MJ, Loew LM
JournalJ Cell Biol
PubMed ID7929557
'Directional cellular locomotion is thought to involve localized intracellular calcium changes and the lateral transport of cell surface molecules. We have examined the roles of both calcium and cell surface glycoprotein redistribution in the directional migration of two murine fibroblastic cell lines, NIH 3T3 and SV101. These cell types exhibit ... More
'Information storage in the nervous system requires transcription triggered by synaptically evoked calcium signals. It has been suggested that translocation of calmodulin into the nucleus, initiated by submembranous calcium transients, relays synaptic signals to CREB. Here we show that in hippocampal neurons, signaling to CREB can be activated by nuclear ... 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
Novel fluorescence sensing methods for high throughput screening.
AuthorsLakowicz JR, Gryczynski I, Gryczynski Z
JournalJ Biomol Screen
PubMed ID10894755
'We describe two new methods of fluorescence sensing for use in high throughput screening (HTS). Modulation sensing transforms analyte-dependent intensity changes into a change in the low-frequency modulation signal. Polarization sensing transforms an intensity change into a change in polarization. Both methods are internally calibrated by using a reference film ... More
Biphasic effects of cytosolic Ca2+ on Ins(1,4,5)P3-stimulated Ca2+ mobilization in hepatocytes.
AuthorsMarshall IC, Taylor CW
JournalJ Biol Chem
PubMed ID8514760
'The increase in cytosolic free Ca2+ concentration that follows mobilization of intracellular Ca2+ stores by inositol 1,4,5-trisphosphate Ins(1,4,5)P3 has been reported to modulate the sensitivity of Ins(1,4,5)P3 receptors. We have examined the effects of cytosolic Ca2+ on Ins(1,4,5)P3-stimulated Ca2+ mobilization in permeabilized hepatocytes. Increasing the free Ca2+ concentration in the ... More
Ca2+-dependent exocytotic pathways in Chinese hamster ovary fibroblasts revealed by a caged-Ca2+ compound.
AuthorsNinomiya Y, Kishimoto T, Miyashita Y, Kasai H
JournalJ Biol Chem
PubMed ID8663485
'Ca2+-dependent exocytosis and endocytosis of Chinese hamster ovary (CHO) fibroblasts were investigated using capacitance measurement and rapid photolysis of a caged-Ca2+ compound, dimethoxynitrophenamine tetrasodium salt. CHO cells exhibited large and fast increases in membrane capacitance (1.9 +/- 1 picofarads, or 13 +/- 7% of total membrane area, mean +/- S.D., ... More
Effects of imperatoxin A on local sarcoplasmic reticulum Ca(2+) release in frog skeletal muscle.
AuthorsShtifman A, Ward CW, Wang J, Valdivia HH, Schneider MF
JournalBiophys J
PubMed ID10920014
'We have investigated the effects of imperatoxin A (IpTx(a)) on local calcium release events in permeabilized frog skeletal muscle fibers, using laser scanning confocal microscopy in linescan mode. IpTx(a) induced the appearance of Ca(2+) release events from the sarcoplasmic reticulum that are approximately 2 s and have a smaller amplitude ... More
Two Ca2+ entry pathways mediate InsP3-sensitive store refilling in guinea-pig colonic smooth muscle.
AuthorsMcCarron JG, Flynn ER, Bradley KN, Muir TC
JournalJ Physiol
PubMed ID10811730
'Sarcolemma Ca2+ influx, necessary for store refilling, was well maintained, over a wide range (-70 to + 40 mV) of membrane voltages, in guinea-pig single circular colonic smooth muscle cells, as indicated by the magnitude of InsP3-evoked Ca2+ transients. This apparent voltage independence of store refilling was achieved by the ... More
Flash photolysis studies of excitation-contraction coupling, regulation, and contraction in smooth muscle.
AuthorsSomlyo AP, Somlyo AV
JournalAnnu Rev Physiol
PubMed ID2184779
'1. Flash photolysis of caged compounds of phenylephrine, inositol 1, 4, 5 trisphosphate (InsP3), GTP gamma S, ATP, and CTP has been successfully used to study excitation-contraction coupling, contractile regulation, and contraction in smooth muscle. Major processes explored with this method were (a) the delay between agonist-receptor interaction and contraction ... More
Time courses of mammalian cell electropermeabilization observed by millisecond imaging of membrane property changes during the pulse.
AuthorsGabriel B, Teissié J
JournalBiophys J
PubMed ID10096909
'Time courses of electropermeabilization were analyzed during the electric field application using a rapid fluorescent imaging system. Exchanges of calcium ions through electropermeabilized membrane of Chinese hamster ovary cells were found to be asymmetrical. Entry of calcium ions during a millisecond pulse occurred on the anode-facing cell hemisphere. Entry through ... More
Subcellular calcium transients visualized by confocal microscopy in a voltage-clamped vertebrate neuron.
AuthorsHernández-Cruz A, Sala F, Adams PR
JournalScience
PubMed ID2154851
'Confocal laser-scanned microscopy and long-wavelength calcium (Ca2+) indicators were combined to monitor both sustained and rapidly dissipating Ca2+ gradients in voltage-clamped sympathetic neurons isolated from the bullfrog. After a brief activation of voltage-dependent Ca2+ channels, Ca2+ spreads inwardly, and reaches the center of these spherical cells in about 300 milliseconds. ... More
Cyclic ADP-ribose-gated Ca2+ release in sea urchin eggs requires an elevated.
AuthorsGuo X, Becker PL
JournalJ Biol Chem
PubMed ID9202011
'Cyclic ADP-ribose (cADPr) has been shown to release intracellular Ca2+ from sea urchin eggs and a variety of vertebrate cell types, although its mechanism of action remains elusive. We employed the caged version of cADPr to study the [Ca2+] transient kinetics in intact sea urchin eggs for insights into how ... More
Fluorescent calcium indicators: subcellular behavior and use in confocal imaging.
AuthorsO'Malley DM, Burbach BJ, Adams PR
JournalMethods Mol Biol
PubMed ID10231797
Inositol trisphosphate is required for the propagation of calcium waves in Xenopus oocytes.
AuthorsDeLisle S, Welsh MJ
JournalJ Biol Chem
PubMed ID1569053
'Stimuli which act through the second messenger inositol 1,4,5-trisphosphate (InsP3) often increase free intracellular Ca2+ concentration ([Ca2+]i) in a localized subcellular area. Actively propagated Ca2+ waves then extend this focal Ca2+ signal to other parts of the cell. To understand how cells may control the spatial distribution of Ca2+, we ... More
Confocal imaging of dendritic Ca2+ transients in hippocampal brain slices during simultaneous current- and voltage-clamp recording.
AuthorsJaffe DB, Brown TH
JournalMicrosc Res Tech
PubMed ID7841500
'Changes in the intracellular Ca2+ concentration ([Ca2+]i) within CA1 hippocampal pyramidal neurons were imaged using confocal laser scanning microscopy in conjunction with Ca(2+)-sensitive fluorescent indicators. The imaging was performed in thick hippocampal brain slices while simultaneously measuring or controlling electrical activity with sharp microelectrodes or whole-cell patch-clamp electrodes. The combination ... More
Submicroscopic calcium signals as fundamental events of excitation--contraction coupling in guinea-pig cardiac myocytes.
AuthorsLipp P, Niggli E
JournalJ Physiol
PubMed ID8730580
'1. Subcellularly localized Ca2+ signals have been proposed to represent elementary events of cardiac Ca2+ signalling (Ca2+ sparks), whereby an individual sarcolemmal L-type Ca2+ channel locally controls opening of a single (or a few) Ca2+ release channels in the sarcoplasmic reticulum (SR). 2. To investigate directly the elementary nature of ... More
Kinetic properties of DM-nitrophen and calcium indicators: rapid transient response to flash photolysis.
AuthorsEscobar AL, Velez P, Kim AM, Cifuentes F, Fill M, Vergara JL
JournalPflugers Arch
PubMed ID9242727
'We describe a high temporal resolution confocal spot microfluorimetry setup which makes possible the detection of fluorescence transients elicited by Ca2+ indicators in response to large (50-200 microM), short duration (< 100 ns), free [Ca2+] transients generated by laser flash photolysis of DM-nitrophen (DM-n; caged Ca2+). The equilibrium and kinetic ... More
Characterization of Ca2+ transients induced by intracellular photorelease of InsP3 in mouse ovarian oocytes.
AuthorsPeres A, Bertollini L, Racca C
JournalCell Calcium
PubMed ID1934036
'Ca2+ transients (measured with Fluo-3) were induced in single mouse ovarian oocytes by photolytic liberation of InsP3. The time course of cytosolic Ca2+ changes induced in this way is composed of distinct phases: upstroke, fast decline, slow declining plateau and fast decline to rest level. All the phases reflect mainly ... More
Compartmentalized and binary behavior of terminal dendrites in hippocampal pyramidal neurons.
AuthorsWei DS, Mei YA, Bagal A, Kao JP, Thompson SM, Tang CM
JournalScience
PubMed ID11567143
'The dendritic arbor of pyramidal neurons is not a monolithic structure. We show here that the excitability of terminal apical dendrites differs from that of the apical trunk. In response to fluorescence-guided focal photolysis of caged glutamate, individual terminal apical dendrites generated cadmium-sensitive all-or-none responses that were subthreshold for somatic ... More
Patch-clamp recording of charge movement, Ca(2+) current, and Ca(2+) transients in adult skeletal muscle fibers
AuthorsWang ZM, Messi ML, Delbono O
JournalBiophys J
PubMed ID10545370
'Intramembrane charge movement (Q), Ca(2+) conductance (G(m)) through the dihydropyridine-sensitive L-type Ca(2+) channel (DHPR) and intracellular Ca(2+) fluorescence (F) have been recorded simultaneously in flexor digitorum brevis muscle fibers of adult mice, using the whole-cell configuration of the patch-clamp technique. The voltage distribution of Q was fitted to a Boltzmann ... More
Polarized expression of Ca2+ channels in pancreatic and salivary gland cells. Correlation with initiation and propagation of [Ca2+]i waves.
AuthorsLee MG, Xu X, Zeng W, Diaz J, Wojcikiewicz RJ, Kuo TH, Wuytack F, Racymaekers L, Muallem S
JournalJ Biol Chem
PubMed ID9188472
'In polarized epithelial cells [Ca2+]i waves are initiated in discrete regions and propagate through the cytosol. The structural basis for these compartmentalized and coordinated events are not well understood. In the present study we used a combination of [Ca2+]i imaging at high temporal resolution, recording of Ca2+-activated Cl- current, and ... 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
Inositol 1,4,5-trisphosphate induced Ca2+ release from chloroquine-sensitive and -insensitive intracellular stores in the intraerythrocytic stage of the malaria parasite P. chabaudi.
AuthorsPassos AP, Garcia CR
JournalBiochem Biophys Res Commun
PubMed ID9535800
'Isolated P. chabaudi parasites were permeabilized with digitonin and the function of intracellular Ca2+ stores was studied using the Ca2+ indicators arsenazo III or Fluo 3-acid in the medium. Addition of the second messenger InsP3 (5 microM) to permeabilized parasites leads to Ca2+ release into the medium, with the mean ... More
Two-photon and UV-laser flash photolysis of the Ca2+ cage, dimethoxynitrophenyl-EGTA-4.
AuthorsDelPrincipe F, Egger M, Ellis-Davies GC, Niggli E
JournalCell Calcium
PubMed ID10191963
'We report efficient two-photon and UV-laser flash photolysis of dimethoxynitrophenyl-EGTA-4 (DMNPE-4), a newly-developed photolabile Ca(2+)-specific chelator. This compound exhibits good two-photon absorption at 705 nm, has a low Mg2+ affinity (approximately 7 mM), a Kd for Ca2+ of 19 nM, a quantum yield of 0.20 and changes its Ca2+ affinity ... More
ATP-induced cytoplasmic calcium mobilization in Bergmann glial cells.
AuthorsKirischuk S, Möller T, Voitenko N, Kettenmann H, Verkhratsky A
JournalJ Neurosci
PubMed ID8613725
'ATP receptor mediated Ca2+ signaling was recorded from Bergmann glial cells in cerebellar slices obtained from mice of different ages (postnatal days 6 to 45). To measure the cytoplasmic concentration of Ca2+ ([Ca2+]in), either individual cells were loaded with the Ca(2+)-sensitive probes using the whole cell patch clamp technique or ... More
The light response of Drosophila photoreceptors is accompanied by an increase in cellular calcium: effects of specific mutations.
AuthorsPeretz A, Suss-Toby E, Rom-Glas A, Arnon A, Payne R, Minke B
JournalNeuron
PubMed ID8011336
'Photoreceptors of dissociated Drosophila retinae were loaded with the fluorescent Ca2+ indicators, fluo-3 and Calcium Green-5N. In fluo-3-loaded, wild-type photoreceptors, a rapid increase in fluorescence (Ca2+ signal) accompanied the light-evoked inward current. Removal of extracellular Ca2+ greatly reduced the Ca2+ signal, indicating Ca2+ influx as its major cause. In Calcium ... More
Confocal laser scanning microscopy used to monitor intracellular Ca2+ changes in hippocampal CA 1 neurons during energy deprivation.
AuthorsGrøndahl T, Langmoen IA
JournalBrain Res
PubMed ID9526045
'An increase in intracellular calcium during cerebral ischemia has been proposed as a common final pathway underlying the events leading to neuronal death. Intracellular calcium has been measured with ion selective electrodes during energy deprivation (ED) in hippocampal slices and with fluorescent techniques in neuronal cultures. In the present study, ... More
Ca2+ sparks and Ca2+ waves in saponin-permeabilized rat ventricular myocytes.
AuthorsLukyanenko V, Gyorke S
JournalJ Physiol
PubMed ID10601490
'1. We carried out confocal Ca2+ imaging in myocytes permeabilized with saponin in ''internal'' solutions containing: MgATP, EGTA and fluo-3 potassium salt. 2. Permeabilized myocytes exhibited spontaneous Ca2+ sparks and waves similar to those observed in intact myocytes loaded with fluo-3 AM. 3. In the presence of ''low'' [EGTA] (0.05 ... More
Fluorometric determination of aminopolycarboxylates using Fluo-3.
AuthorsKawasaki N, Lee YC, Hashimoto O, Yamamoto M, Kawanishi T, Hayakawa T
JournalAnal Biochem
PubMed ID10334851
Methods for calibration of fluorescent calcium indicators in skeletal muscle fibers.
AuthorsWesterblad H, Allen DG
JournalBiophys J
PubMed ID8011925
The many ways to cross the plasma membrane.
AuthorsStephens DJ, Pepperkok R
JournalProc Natl Acad Sci U S A
PubMed ID11274366
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
Intracellular ionized calcium.
AuthorsJune CH, Rabinovitch PS
JournalMethods Cell Biol
PubMed ID7861962
Intercellular calcium signaling and flash photolysis of caged compounds. A sensitive method to evaluate gap junctional coupling.
AuthorsLeybaert L, Sanderson MJ
JournalMethods Mol Biol
PubMed ID11218662
Practical aspects of measuring [Ca2+] with fluorescent indicators.
AuthorsKao JP
JournalMethods Cell Biol
PubMed ID8201975
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
Phosphorylation-independent modulation of L-type calcium channels by magnesium-nucleotide complexes.
AuthorsO'Rourke B, Backx PH, Marban E
JournalScience
PubMed ID1321495
Free magnesium ions and magnesium-nucleotide complexes can exert opposite effects on many fundamental cellular processes. Although increases in the intracellular concentration of magnesium ions inhibit the L-type calcium current in heart cells, magnesium-adenosine triphosphate complexes (MgATP) would be expected to increase the current by promoting channel phosphorylation. Rapid increases in ... More
Ca2+ flux through promiscuous cardiac Na+ channels: slip-mode conductance.
AuthorsSantana LF, Gómez AM, Lederer WJ
JournalScience
PubMed ID9461434
The tetrodotoxin-sensitive sodium ion (Na+) channel is opened by cellular depolarization and favors the passage of Na+ over other ions. Activation of the beta-adrenergic receptor or protein kinase A in rat heart cells transformed this Na+ channel into one that is promiscuous with respect to ion selectivity, permitting calcium ions ... More
Calcium buffering capacity of neuronal cell cytosol measured by flash photolysis of calcium buffer NP-EGTA.
AuthorsFleet A, Ellis-Davies G, Bolsover S
JournalBiochem Biophys Res Commun
PubMed ID9784424
N1E-115 mouse neuroblastoma cells were injected with a calcium buffer/indicator solution to allow both ratiometric measurement of free calcium concentration and the release of calcium ions upon UV flash. The solution contained sulforhodamine, a marker dye used to estimate the volume injected; fluo-3, a calcium indicator, and NP-EGTA, a high ... More
Control of mitochondrial matrix calcium: studies using fluo-3 as a fluorescent calcium indicator.
AuthorsSaavedra-Molina A, Uribe S, Devlin TM
JournalBiochem Biophys Res Commun
PubMed ID2310386
Fluo-3, a fluorescent Ca2+ indicator, is sequestered by isolated rat liver mitochondria and is an effective probe for evaluating the concentration and kinetics of change of mitochondrial matrix ionized calcium ([Ca2+]m) under a variety of conditions. At the wavelengths employed, there is no significant interference by auto-fluorescence. There is an ... More
Activity-induced potentiation of developing neuromuscular synapses.
AuthorsWan J, Poo M
JournalScience
PubMed ID10481007
Electrical activity plays a critical role in shaping the structure and function of synaptic connections in the nervous system. In Xenopus nerve-muscle cultures, a brief burst of action potentials in the presynaptic neuron induced a persistent potentiation of neuromuscular synapses that exhibit immature synaptic functions. Induction of potentiation required an ... More
The effect of pH on rate constants, ion selectivity and thermodynamic properties of fluorescent calcium and magnesium indicators.
AuthorsLattanzio FA, Bartschat DK
JournalBiochem Biophys Res Commun
PubMed ID2043105
Fluorescent calcium indicators fluo-3, fura-2 and indo-1, and fluorescent magnesium indicators mag-fura-2 (FURAPTRA) and mag-indo-1 were evaluated for the effects of pH on their association and dissociation rates, ion selectivity and thermodynamic properties. Calcium indicator affinities for Ca and Mg were reduced and the discrimination between Ca and Mg decreased ... More
Cytosolic calcium responses induced by photolytic release of 1,4,5-inositol trisphosphate in single human fibroblasts.
AuthorsPeres A, Racca C, Bertollini L, Sturani E
JournalBiochim Biophys Acta
PubMed ID2009312
We have used the whole cell technique to microinject human fibroblasts with either 1,4,5-inositol trisphosphate (InsP3) or 'caged' InsP3, in order to study the mechanisms of transmembrane signalling related to mitogenic stimulations. Cytosolic Ca2+ elevations in response to 1,4,5 InsP3 diffusing from the patch pipette were difficult to detect, while ... More
Confocal microscopy of fertilization-induced calcium dynamics in sea urchin eggs.
AuthorsStricker SA, Centonze VE, Paddock SW, Schatten G
JournalDev Biol
PubMed ID1730391
Although confocal microscopy has typically been utilized in studies of fixed specimens, its potential for exploring dynamic processes in living cells is rapidly being realized. In this report, confocal laser scanning microscopy is used to analyze the calcium wave that occurs following fertilization in living sea urchin eggs microinjected with ... More
Chemical transformations in individual ultrasmall biomimetic containers.
AuthorsChiu DT, Wilson CF, Ryttsén F, Strömberg A, Farre C, Karlsson A, Nordholm S, Gaggar A, Modi BP, Moscho A, Garza-López RA, Orwar O, Zare RN
JournalScience
PubMed ID10082457
Individual phospholipid vesicles, 1 to 5 micrometers in diameter, containing a single reagent or a complete reaction system, were immobilized with an infrared laser optical trap or by adhesion to modified borosilicate glass surfaces. Chemical transformations were initiated either by electroporation or by electrofusion, in each case through application of ... More
Low-frequency modulation sensors using nanosecond fluorophores.
AuthorsLakowicz JR, Castellano FN, Dattelbaum JD, Tolosa L, Rao G, Gryczynski I
JournalAnal Chem
PubMed ID9868909
We describe a new approach to fluorescence sensing based on a mixture of fluorophores, one of which is sensitive to the desired analyte. If a long-lifetime analyte-insensitive fluorophore is mixed with a short-lifetime analyte-sensitive fluorophore, the modulation of the emission at conveniently low frequencies becomes equal to the fractional fluorescence ... More
Local calcium transients triggered by single L-type calcium channel currents in cardiac cells.
AuthorsLópez-López JR, Shacklock PS, Balke CW, Wier WG
JournalScience
PubMed ID7754383
Excitation-contraction coupling was studied in mammalian cardiac cells in which the opening probability of L-type calcium (Ca2+) channels was reduced. Confocal microscopy during voltage-clamp depolarization revealed distinct local transients in the concentration of intracellular calcium ions ([Ca2+]i). When voltage was varied, the latency to occurrence and the relative probability of ... 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
Nicotinic cholinergic signaling in hippocampal astrocytes involves calcium-induced calcium release from intracellular stores.
AuthorsSharma G, Vijayaraghavan S,
JournalProc Natl Acad Sci U S A
PubMed ID11259680
In this report we provide evidence that neuronal nicotinic acetylcholine receptors (nAChRs) are present on hippocampal astrocytes and their activation produces rapid currents and calcium transients. Our data indicate that these responses obtained from astrocytes are primarily mediated by an AChR subtype that is functionally blocked by alpha-bungarotoxin (alpha Bgt) ... More
IP3-dependent nuclear Ca2+ signalling in the mammalian heart.
AuthorsZima AV, Bare DJ, Mignery GA, Blatter LA,
JournalJ Physiol
PubMed ID17761776
In cardiac myocytes the type-2 inositol 1,4,5-trisphosphate receptor (IP(3)R2) is the predominant isoform expressed. The IP(3)R2 channel is localized to the SR and to the nuclear envelope. We studied IP(3)-dependent nuclear Ca(2+) signals ([Ca(2+)](Nuc)) in permeabilized atrial myocytes and in isolated cardiac nuclei. In permeabilized myocytes IP(3) (20 microm) and ... More
Range of messenger action of calcium ion and inositol 1,4,5-trisphosphate.
AuthorsAllbritton NL, Meyer T, Stryer L
JournalScience
PubMed ID1465619
The range of messenger action of a point source of Ca2+ or inositol 1,4,5-trisphosphate (IP3) was determined from measurements of their diffusion coefficients in a cytosolic extract from Xenopus laevis oocytes. The diffusion coefficient (D) of [3H]IP3 injected into an extract was 283 microns 2/s. D for Ca2+ increased from ... More
Dendritic spines for neuroprotection: a hypothesis.
AuthorsSegal M
JournalTrends Neurosci
PubMed ID8592749
Ever since their first description in neurons, dendritic spines could be visualized only in fixed tissue, using high-power light and electron microscopy. Recent studies have been able to measure the free intracellular Ca2+ concentration ([Ca2+]i) in dendritic spines of live neurons, and the results suggest that the spine is an ... More
Inhibition by Ca2+ of inositol trisphosphate-mediated Ca2+ liberation: a possible mechanism for oscillatory release of Ca2+.
AuthorsParker I, Ivorra I
JournalProc Natl Acad Sci U S A
PubMed ID2296584
Light-flash photolysis of caged inositol 1,4,5-trisphosphate (InsP3) was used to generate reproducible transients of free InsP3 in Xenopus oocytes, and the resulting liberation of Ca2+ from intracellular stores was monitored by recording Ca2+-activated membrane currents and by use of the fluorescent Ca2+ indicator fluo-3. InsP3-mediated Ca2+ release was inhibited by ... More
Kinetics of calcium binding to fluo-3 determined by stopped-flow fluorescence.
AuthorsEberhard M, Erne P
JournalBiochem Biophys Res Commun
PubMed ID2775268
The kinetics of Ca2+ dissociation from fluo-3 was measured using stopped flow fluorimetry. Analysis of dissociation revealed, in contrast to other commonly used fluorescent Ca2+ indicators, a biexponential behaviour with two distinct dissociation rates of 550 s-1 and 200 s-1 at physiological pH and room temperature. The dissociation rate constant ... More
Localized all-or-none calcium liberation by inositol trisphosphate.
AuthorsParker I, Ivorra I
JournalScience
PubMed ID2237441
Laser confocal microscopy was used to monitor calcium ion (Ca2+) liberation from highly localized (micrometer) regions of intact Xenopus oocytes in response to photo-released inositol 1,4,5-trisphosphate (InsP3). Local Ca2+ release varied in an all-or-none manner with increasing amount of InsP3, in contrast to signals recorded from larger areas, which grew ... More
Demonstration of calcium uptake and release by sea urchin egg cortical endoplasmic reticulum.
AuthorsTerasaki M, Sardet C
JournalJ Cell Biol
PubMed ID1955454
The calcium indicator dye fluo-3/AM was loaded into the ER of isolated cortices of unfertilized eggs of the sea urchin Arbacia punctulata. Development of the fluorescent signal took from 8 to 40 min and usually required 1 mM ATP. The signal decreased to a minimum level within 30 s after ... 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
Detection of Ca(2+)-transients elicited by flash photolysis of DM-nitrophen with a fast calcium indicator.
AuthorsEscobar AL, Cifuentes F, Vergara JL
JournalFEBS Lett
PubMed ID7758592
A confocal spot detection optical setup was used to record fluorescence signals in response to calcium pulses, elicited by flash photolysis of DM-nitrophen, with the calcium indicators CaOrange-5N and Fluo-3. Our results yield the following conclusions: [Ca2+] changes are almost perfect spikes at pCa 9 and broader transients followed by ... More
Photo-released intracellular Ca2+ evokes reversible mechanical responses in supporting cells of the guinea-pig organ of Corti.
AuthorsDulon D, Blanchet C, Laffon E
JournalBiochem Biophys Res Commun
PubMed ID8024570
Loaded under whole cell patch-clamp configuration, the caged Ca2+ molecule DM-nitrophen was used to increase [Ca2+]i rapidly and reversibly in isolated Deiters cells of the organ of Corti. Photolysis of DM-nitrophen increased [Ca2+]i from resting concentrations of 20-50 nM to values above microM, as measured with the fluorescent indicator Fluo-3. ... More
Unique inactivation properties of NAADP-sensitive Ca2+ release.
AuthorsGenazzani AA, Empson RM, Galione A
JournalJ Biol Chem
PubMed ID8662773
Ca2+ mobilization from intracellular stores constitutes an important mechanism for generating cytoplasmic Ca2+ signals. Inositol trisphosphate (InsP3) and ryanodine receptors are the two families of intracellular Ca2+ release channels that have been identified, which may be regulated by separate intracellular messengers, InsP3, and cyclic adenosine 5'-diphosphate ribose, respectively. A third ... More
Effects of oxidants on properties of fluorescent calcium indicators.
AuthorsSarvazyan N, Swift L, Martinez-Zaguilan R
JournalArch Biochem Biophys
PubMed ID9466830
An increasing number of studies use calcium-sensitive fluorescent dyes to address the relationship between elevated levels of intracellular calcium and free-radical-mediated damage in a variety of pathophysiological phenomena. The present study evaluates the effects of reactive oxygen species on the spectral properties of widely used calcium probes such as Fura-2 ... More