Abstract：
:The fluo family of indicators is frequently used in studying Ca(2+) physiology; however, choosing which fluo indicator to use is not obvious. Indicator properties are typically determined in well-defined aqueous solutions. Inside cells, however, the properties can change markedly. We have characterized each of three fluo variants (fluo-2MA, fluo-3 and fluo-4) in two forms-the acetoxymethyl (AM) ester and the K(+) salt. We loaded indicators into rat ventricular myocytes and used confocal microscopy to monitor depolarization-induced fluorescence changes and fractional shortening. Myocytes loaded with the indicator AM esters showed significantly different Ca(2+) transients and fractional shortening kinetics. Loading the K(+) salts via whole-cell patch-pipette eliminated differences between fluo-3 and fluo-4, but not fluo-2MA. Cells loaded with different indicator AM esters showed different staining patterns-suggesting differential loading into organelles. Ca(2+) dissociation constants (K(d,Ca)), measured in protein-rich buffers mimicking the cytosol were significantly higher than values determined in simple buffers. This increase in K(d,Ca) (decrease in Ca(2+) affinity) was greatest for fluo-3 and fluo-4, and least for fluo-2MA. We conclude that the structurally-similar fluo variants differ with respect to cellular loading, subcellular compartmentalization, and intracellular Ca(2+) affinity. Therefore, judicious choice of fluo indicator and loading procedure is advisable when designing experiments.
journal_name
Cell Calciumjournal_title
Cell calciumauthors
Hagen BM,Boyman L,Kao JP,Lederer WJdoi
10.1016/j.ceca.2012.05.010subject
Has Abstractpub_date
2012-08-01 00:00:00pages
170-81issue
2eissn
0143-4160issn
1532-1991pii
S0143-4160(12)00106-6journal_volume
52pub_type
杂志文章相关文献
CELL CALCIUM文献大全abstract：:Ca2+ influx into secretory cells of the intact salivary gland of the blowfly Calliphora erythrocephala elicited by the agonist 5-hydroxytryptamine (5-HT) or the Ca2+ uptake inhibitor thapsigargin was studied by using Fura-2 and digital fluorescence imaging and by recordings of the transepithelial potential. Applicatio...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/s0143-4160(98)90074-4
更新日期：1998-01-01 00:00:00
abstract：:In the brain, classical (canonical) transient receptor potential (TRPC) channels are thought to be involved in different aspects of neuronal development. We investigated the developmental expression profile of TRPC channels in rat cerebellum during the first 6 weeks after birth. TRPC3 expression is significantly up-re...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2006.11.002
更新日期：2007-07-01 00:00:00
abstract：:We have used single cell fluorescence imaging techniques to examine how functional properties of the caffeine-sensitive Ca(2+) store change during differentiation of a sub-population of caffeine-sensitive SH-SY5Y cells. Application of caffeine (30 mM) 1-10.5 min after a 'priming' depolarisation pulse of 55 mM K(+) rev...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2006.06.007
更新日期：2007-03-01 00:00:00
abstract：:Although store-operated calcium entry (SOCE) was identified more that two decades ago, understanding the molecular mechanisms that regulate and mediate this process continue to pose a major challenge to investigators in this field. Thus, there has been major focus on determining which of the models proposed for this m...
journal_title：Cell calcium
pub_type： 杂志文章,评审
doi：10.1016/j.ceca.2007.01.013
更新日期：2007-08-01 00:00:00
abstract：:We report new fluorescent calcium indicators based on fluo-4. Attachment of a carboxamide or methylenecarboxamide moiety to the BAPTA chelator portion of fluo-4 allowed for the attachment of dextrans, protein-reactive moieties, and biotin. In particular, a high affinity fluo-4 dextran conjugate was prepared and shown ...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2004.05.002
更新日期：2004-12-01 00:00:00
abstract：:Photolysis of caged calcium (Nitr5, Calbiochem) can be used to study calcium dependent processes such as excitation-contraction coupling and muscular mechanics. Expensive high energy light sources are routinely used for UV light exposure, but this study describes an alternative low cost xenon flash unit constructed in...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/0143-4160(94)90017-5
更新日期：1994-05-01 00:00:00
abstract：:Apparent calcium stability constants and the purity of Ca-chelating agents were evaluated using Ca(2+)-selective electrodes by the double-log optimization method [Oiki S. Yamamoto T. Okada Y. (1994) Cell Calcium, 15, 199-208]. The method was amended to allow evaluation of the free Ca2+ concentrations contaminating ele...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/0143-4160(94)90060-4
更新日期：1994-03-01 00:00:00
abstract：:The regulation of cytosolic-free calcium concentration of smooth-muscle and endothelial cells was mainly studied on cultured cells where the cross talk between these two coupled cell types is lost. In the present study, the cytosolic-free calcium concentration in the endothelial and the smooth-muscle cells was examine...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1054/ceca.2001.0233
更新日期：2001-10-01 00:00:00
abstract：:The tissue/medium distribution of the nonmetabolized glucose analog [14C]-3-O-methyl-D-glucose was measured in pigeon erythrocytes and related to changes in 45Ca uptake and efflux, total calcium content and ATP levels. Sugar transport was not affected by changes in external Ca2+. However, both sugar and 45Ca influx we...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/0143-4160(82)90004-5
更新日期：1982-08-01 00:00:00
abstract：:In skeletal muscle excitation-contraction (EC) coupling the sarcolemmal L-type Ca(2+) channel or 1,4-dihydropyridine receptor (DHPR) transduces the membrane depolarization signal to the sarcoplasmic Ca(2+) release channel RyR1 via protein-protein interaction. While it is evident that the pore-forming and voltage-sensi...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2010.04.003
更新日期：2010-06-01 00:00:00
abstract：:The non-selective second messenger-gated cation channel TRPC6 (transient receptor potential canonical 6) is activated by diacylglycerols (DAG) in a PKC-independent manner and plays important roles in a variety of physiological processes and diseases. In order to facilitate novel therapies, the development of potent in...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2018.12.009
更新日期：2019-03-01 00:00:00
abstract：:While genetically encoded Ca(2+) indicators (GECIs) allow Ca(2+) imaging in model organisms, the gene expression is often under the control of a single promoter that may drive expression beyond, the cell types of interest. To enable more cell-type specific targeting, GECIs can be brought under the, control of the inte...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2011.10.006
更新日期：2012-01-01 00:00:00
abstract：:Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by selective death of motor neurons. Mutations in Cu/Zn superoxide dismutase-1 (SOD1) cause familial ALS but the molecular mechanisms whereby these mutations induce motor neuron death remain controversial. Here, we show that stable overex...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2011.02.004
更新日期：2011-04-01 00:00:00
abstract：:MiR-204 is expressed in vascular smooth muscle cells (VSMC). However, its role in VSMC contraction is not known. We determined if miR-204 controls VSMC contractility and blood pressure through regulation of sarcoplasmic reticulum (SR) calcium (Ca2+) release. Systolic blood pressure (SBP) and vasoreactivity to VSMC con...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2019.03.006
更新日期：2019-06-01 00:00:00
abstract：:This investigation concentrates on the change in Ca(2+) concentration ([Ca(2+)]) caused by ryanodine in U373 MG cells. This cell type from a human astrocytoma is a unique cellular model because it only expresses the type 3 ryanodine receptor (RyR3), which is generally the least abundant isoform. In the presence of phy...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2004.02.004
更新日期：2004-11-01 00:00:00
abstract：:Calcium dependence of bleb formation and cell death was evaluated in rat hepatocytes following ATP depletion by metabolic inhibition with KCN and iodoacetate ('chemical hypoxia'). Cytosolic free Ca2+ was measured in single cells by ratio imaging of Fura-2 fluorescence using multiparameter digitized video microscopy. C...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/0143-4160(88)90004-8
更新日期：1988-12-01 00:00:00
abstract：:Using whole cell patch clamp recordings on unfertilized eggs of the ascidian Ciona intestinalis, we are able to detect ryanodine receptors within the oocytes. Our approach is based on measurements of the voltage-activated inward calcium currents. Two types of Ca2+ currents have been described on the oocyte membrane of...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/s0143-4160(97)90033-6
更新日期：1997-02-01 00:00:00
abstract：:TRP family of proteins are components of unique cation channels that are activated in response to diverse stimuli ranging from growth factor and neurotransmitter stimulation of plasma membrane receptors to a variety of chemical and sensory signals. This review will focus on members of the TRPC sub-family (TRPC1-TRPC7)...
journal_title：Cell calcium
pub_type： 杂志文章,评审
doi：10.1016/j.ceca.2006.08.011
更新日期：2006-11-01 00:00:00
abstract：:Cyclic AMP regulates the late step of Ca²+-dependent exocytosis in many secretory cells through two major mechanisms: a protein kinase A-dependent and a cAMP-GEF/Epac-dependent pathway. We designed a protocol to characterize the role of these two cAMP-dependent pathways on the Ca²+ sensitivity and kinetics of regulate...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2010.12.005
更新日期：2011-02-01 00:00:00
abstract：:The Ca2+-sensitive dye quin-2 was used to measure the cytosolic free Ca2+ (Cai2+) in suspensions of ventricular myocytes isolated from cat and rat ventricles. Following an isolation procedure that was similar for both species, the cells were loaded with quin-2 AM (25 microM) for 30 min at 37 degrees C. After two washe...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/0143-4160(87)90002-9
更新日期：1987-08-01 00:00:00
abstract：:The measurement of intracellular calcium ion concentrations [( Ca2+]i) in single living cells using quantitative fluorescence microscopy draws from a diverse set of disciplines, including cellular biology, optical physics, statistics and computer science. Over the last few years, we have devised and built a number of ...
journal_title：Cell calcium
pub_type： 杂志文章,评审
doi：10.1016/0143-4160(90)90068-6
更新日期：1990-02-01 00:00:00
abstract：:The effect of furosemide and ouabain on the intracellular concentration of Ca2+ was studies in myocardial cell cultures using Fura-2, a fluorescent agent, as an intracellular Ca2+ indicator. Introduction of 200 microM ouabain to the cultured cells increased the intracellular calcium concentration from an average of 23...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/0143-4160(95)90004-7
更新日期：1995-08-01 00:00:00
abstract：:Endothelin-1 (ET-1), a potent vasoactive peptide with a pathogenic role in vascular diseases, has been shown to induce the activation of ERK1/2, PKB and the expression of a transcriptional regulator, the early growth response 1 (Egr-1), key mediators of hypertrophic and proliferative responses in vascular smooth muscl...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2013.10.002
更新日期：2013-12-01 00:00:00
abstract：:The strength of the heart beat depends on the amplitude and time course of the transient increase in [Ca(2+)] in the myocytes with each cycle. [Na(+)](i) modulates cardiac contraction through its effect on the Ca(2+) flux through the Na/Ca exchanger. Cardiac excitation-contraction coupling has been postulated to occur...
journal_title：Cell calcium
pub_type： 杂志文章,评审
doi：10.1016/j.ceca.2004.01.014
更新日期：2004-06-01 00:00:00
abstract：:Here we report on the feasibility of using replication deficient adenoviruses to modify signal transduction systems in epithelia. We constructed two viruses, one expressing a dominant negative mutant of the alpha-subunit of Gq (Ad-EF1-dnG alpha q) and the other expressing the wild-type alpha-subunit of Gq (Ad-EF1-wtG ...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/s0143-4160(98)90077-x
更新日期：1998-08-01 00:00:00
abstract：:In addition to its role in artery contraction, Rho kinase (ROCK) is reported to be involved in the Ca(2+) response to vasoconstrictor agonist in rat aorta. However the signaling pathway mediated by ROCK had not been investigated so far and it was not known whether ROCK also contributed to Ca(2+) signaling in cultured ...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2012.07.002
更新日期：2012-12-01 00:00:00
abstract：:In non-excitable cells, the inositol 1,4,5-trisphosphate receptor (IP3R) is an intracellular Ca2+ channel which plays a major role in Ca2+ signalling. Three isoforms of IP3R have been identified (IP3R-1, IP3R-2 and IP3R-3) and most cell types express different proportions of each isoform. The differences between the p...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2007.01.002
更新日期：2007-09-01 00:00:00
abstract：:A procedure for calibration of fluorescence signals from adult rat heart cells loaded with the -AM ester of fura-2 is described. Calibration is complicated by dye compartmentation and potentially incomplete dye hydrolysis. These problems were overcome by subtracting from fluorescence transients the non-cytosolic (mito...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/s0143-4160(98)90050-1
更新日期：1998-10-01 00:00:00
abstract：:Ryanodine receptors plays a crucial role in skeletal muscle excitation-contraction coupling by releasing calcium ions required for muscle contraction from the sarcoplasmic reticulum. At least three phenotypes associated with more than 100 RYR1 mutations have been identified; in order to elucidate possible pathophysiol...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/j.ceca.2008.10.001
更新日期：2009-02-01 00:00:00
abstract：:Calbindin D28K (formerly known as vitamin D-dependent calcium binding protein and referred to here as calbindin) is found in a wide variety of tissues, but only in certain cells within those tissues. Apart from its ability to bind calcium, nothing is known about its function in these cells. To investigate its role we ...
journal_title：Cell calcium
pub_type： 杂志文章
doi：10.1016/0143-4160(93)90055-b
更新日期：1993-04-01 00:00:00
