Simultaneous measurement of intracellular pH and Ca2+ in insulin-secreting cells by spectral imaging microscopy.
AuthorsMartínez-Zaguilán R,Gurulé MW,Lynch RM
JournalThe American journal of physiology
PubMed ID8967445
Measurement of intracellular pH.
AuthorsBoyer MJ, Hedley DW
JournalMethods Cell Biol
PubMed ID7532259
Fluorescent indicators of ion concentrations.
AuthorsTsien RY
JournalMethods Cell Biol
PubMed ID2538708
Comparison of spectrum-shifting intracellular pH probes 5'(and 6')-carboxy-10-dimethylamino-3-hydroxyspiro[7H-benzo[c]xanthene-7, 1'(3'H)-isobenzofuran]-3'-one and 2',7'-biscarboxyethyl-5(and 6)-carboxyfluorescein.
AuthorsOwen CS
JournalAnal Biochem
PubMed ID1514696
'The dyes carboxy-SNARF-1 and BCECF are fluorescent probes of intracellular pH that exhibit changes in spectral shape upon proton binding which allow one to use measurements of fluorescence at two or more wavelengths in order to measure pH without artifacts associated with variability in dye loading, etc. In evaluating these ... More
Profiling pH changes in the electrospray plume.
AuthorsZhou S, Prebyl BS, Cook KD
JournalAnal Chem
PubMed ID12380808
'Laser-induced fluorescence spectrometry is used to profile pH changes as droplets evaporate in an electrospray plume by measuring emission spectra of 2(or 4)-[10-(dimethylamino)-3-oxo-3H-benzo[c]xanthene-7-yl]-benzenedicarboxylic acid (carboxy SNARF-1), a pH-sensitive fluorescent dye. The observed pH changes depend on initial droplet pH and polarity. In some instances, small or negligible changes of pH ... More
Real-time multi-wavelength fluorescence imaging of living cells.
AuthorsMorris SJ
JournalBiotechniques
PubMed ID2331370
'We describe a new real-time fluorescence video microscope design for capturing intensified images of cells containing dual wavelength "ratio" dyes or multiple dyes. The microscope will perform real-time capture of two separate fluorescence emission images simultaneously, improving the time resolution of spatial distribution of fluorescence to video frame rates (30 ... More
SNARF-1 as an intracellular pH indicator in laser microspectrofluorometry: a critical assessment.
AuthorsSeksek O, Henry-Toulmé N, Sureau F, Bolard J
JournalAnal Biochem
PubMed ID2042742
'The use of SNARF-1-AM (seminaphtorhodafluor-1-acetoxymethylester) to measure the internal pH of a single living cell by laser microspectrofluorometry has been analyzed with a lymphocyte murine B cell line A20. After incubation of the cells at 37 degrees C in the presence of 10 microM SNARF-1-AM, the internal concentration of SNARF-1 ... More
A comparison of the effects of intracellular and extracellular pH on contraction in isolated rat portal vein.
AuthorsTaggart M, Austin C, Wray S
JournalJ Physiol
PubMed ID8021835
'1. The effects of changes in extracellular and intracellular pH on spontaneous contractile activity in isolated rat portal vein have been investigated. 2. Small strips of portal vein were loaded with the pH-sensitive fluorophore carboxy-SNARF and intracellular pH (pHi) and contraction were measured simultaneously at 37 degrees C. The tissue ... More
Stepwise cytoskeletal polarization as a series of checkpoints in innate but not adaptive cytolytic killing.
AuthorsWulfing C, Purtic B, Klem J, Schatzle JD
JournalProc Natl Acad Sci U S A
PubMed ID12802007
'Cytolytic killing is a major effector mechanism in the elimination of virally infected and tumor cells. The innate cytolytic effectors, natural killer (NK) cells, and the adaptive effectors, cytotoxic T cells (CTL), despite differential immune recognition, both use the same lytic mechanism, cytolytic granule release. Using live cell video fluorescence ... More
Occurrence of intracellular pH transients during spontaneous contractions in rat uterine smooth muscle.
AuthorsTaggart MJ, Wray S
JournalJ Physiol
PubMed ID8145141
'1. Intracellular pH (pHi) and contraction have been measured simultaneously in isolated longitudinal rat myometrium. Tissue was loaded with the pH-sensitive fluorophore carboxy-SNARF and superfused with oxygenated, buffered salt solution at 37 degrees C and pH 7.4. 2. Small (0.04 pH unit) transient intracellular acidifications followed each spontaneous uterine contraction. ... More
Simultaneous measurement of intracellular pH and contraction in uterine smooth muscle.
AuthorsTaggart M, Wray S
JournalPflugers Arch
PubMed ID8351202
'Changes in intracellular pH (pHi) are thought to produce large changes in force production in the uterus. There have however, been no simultaneous measurements of pHi and force in the uterus and therefore no direct information is available about the relation between the two. We have used carboxy-SNARF (a pH-sensitive ... More
Spectral and photophysical studies of benzo[c]xanthene dyes: dual emission pH sensors.
AuthorsWhitaker JE, Haugland RP, Prendergast FG
JournalAnal Biochem
PubMed ID1862936
'A series of fluorescent, long-wavelength, benzo[c]-xanthene dyes has been characterized for pH measurement in both excitation and emission ratio applications. The two general classes of these indicators are seminaphthofluoresceins (SNAFLs) and seminaphthorhodafluors (SNARFs) which are substituted at the 10-position with oxygen or nitrogen, respectively. These probes show separate emissions from ... More
Characterization of dual-wavelength seminaphthofluorescein and seminapthorhodafluor dyes for pH sensing under high hydrostatic pressures.
AuthorsSalerno M, Ajimo JJ, Dudley JA, Binzel K, Urayama P
JournalAnal Biochem
PubMed ID17274941
'Hydrostatic pressure is an important physical parameter in biology, with pressures in the few-hundred-atm range having significant effects on cellular morphology, metabolism, and viability. To ensure valid results when studying pressure effects using fluorescence spectroscopy and imaging methods, metabolic probes need to be characterized for high-pressure use. Of interest is ... More
Cytolysin-dependent evasion of lysosomal killing.
AuthorsHåkansson A, Bentley CC, Shakhnovic EA, Wessels MR
JournalProc Natl Acad Sci U S A
PubMed ID15795386
'Local host defenses limit proliferation and systemic spread of pathogenic bacteria from sites of mucosal colonization. For pathogens such as streptococci that fail to grow intracellularly, internalization and killing by epithelial cells contribute to the control of bacterial growth and dissemination. Here, we show that group A Streptococcus (GAS), the ... More
Optical methods to measure membrane transport processes.
AuthorsVerkman AS
JournalJ Membr Biol
PubMed ID8606370
Fluorescent indicators for intracellular pH.
AuthorsHan J, Burgess K,
JournalChem Rev
PubMed ID19831417
This review is about intracellular pH sensors, includingsmall fluorescent organic molecules, nanoparticles, andfluorescent proteins, e.g., GFP. It focuses on their preparations, photophysical properties, and advantages/disadvantagesfor intracellular pH measurements. The discussion is limitedto fluorescent indicators that have been applied to measureintracellular pH values since 1980. ... More
Synthesis and photophysical properties of new fluorinated benzo[c]xanthene dyes as intracellular pH indicators.
AuthorsLiu J, Diwu Z, Leung WY
JournalBioorg Med Chem Lett
PubMed ID11677123
Two new fluorinated benzo[c]xanthene dyes were synthesized by reaction of fluorinated 1,6-dihydroxynaphthalenes with 2,4- (and 2,5)-dicarboxy-3'-dimethylamino-2'-hydroxybenzophenone. The two critical fluorinated 1,6-dihydroxynaphthalene intermediates were prepared via a regioselective route. The fluorinated benzo[c]xanthene dyes exhibit desired lower pK(a) values (6.4 and 7.2, respectively) than their parent compound (pK(a)=7.5) while the pH-dependent dual-emission ... 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
Changes in intracellular pH close to term and their possible significance to labour.
AuthorsParratt JR, Taggart MJ, Wray S
JournalPflugers Arch
PubMed ID8594535
The contractile pattern of uterine smooth muscle is markedly altered by even modest manipulations of intracellular pH (pHi); an acidification can abolish contractions, while alkalinization increases contractility. As at the end of gestation the uterus changes from being relatively quiescent to highly contractile, we investigated whether pHi changes with pregnancy ... More
A method for recording intracellular [Ca2+] transients in cardiac myocytes using calcium green-2.
AuthorsSpencer CI, Berlin JR
JournalPflugers Arch
PubMed ID7491285
Calcium green-2 (Ca green) is a non-ratiometric fluorescent Ca2+ indicator with an affinity for Ca2+ (dissociation constant Kd = 3 microM) that is lower than more commonly used indicators such as fura-2 and fluo-3. This low Ca2+ affinity, coupled with a high quantum yield, allows cells to be loaded with ... More
Measurement of intracellular pH using flow cytometry with carboxy-SNARF-1.
AuthorsWieder ED, Hang H, Fox MH
JournalCytometry
PubMed ID8287734
The new intracellular pH (pH(i)) dye carboxy-seminaphthorhodafluor (SNARF-1) was compared to the established dye 2,3-dicyanohydroquinone (DCH) using flow cytometry. Both dyes give high-resolution pH(i) measurements. SNARF-1 remains trapped within cells much longer than DCH, so that pH(i) can be monitored during and after treatments with chemicals or hyperthermia. The toxicity ... More
Using LysoSensor Yellow/Blue DND-160 to sense acidic pH under high hydrostatic pressures.
AuthorsDePedro HM, Urayama P,
JournalAnal Biochem
PubMed ID18976626
LysoSensor Yellow/Blue DND-160, a dual-wavelength fluorophore commonly used for sensing pH in acidic organelles, possesses solvatochromic behavior believed to originate from an intramolecular charge transfer (ICT). Given this, we investigated whether DND-160 can be used for acidic pH sensing under hydrostatic pressures up to 510 atm, a range suitable for ... More
Dynamic on-column pH monitoring in capillary electrophoresis: application to volume-limited outlet vials.
AuthorsTimperman A, Tracht SE, Sweedler JV
JournalAnal Chem
PubMed ID8694265
With capillary electrophoresis, buffer pH must be constant to achieve consistent migration times. Irreproducible separations have been attributed to pH changes due to water hydrolysis in the inlet/outlet vials. A method of measuring the pH of the electrolyte on-column is described that uses wavelength-resolved fluorescence detection. C.SNARF-1 is a fluorescent ... More
Time-resolved fluorescence microscopy could correct for probe binding while estimating intracellular pH.
AuthorsSrivastava A, Krishnamoorthy G
JournalAnal Biochem
PubMed ID9212865
Estimation of intracellular pH by fluorescence ratiometry overcomes many of the limitations such as variations in the pathlength of observation and concentration of the probe, light scattering, and photobleaching. However, binding of probes to membranes and macromolecules is generally not taken into account. By using time-resolved fluorescence microscopy on a ... More
Optical measurements of pH using fluorescence lifetimes and phase-modulation fluorometry.
AuthorsSzmacinski H, Lakowicz JR
JournalAnal Chem
PubMed ID8368522
We measured the pH-dependent fluorescence decay times of the seminaphthofluoresceins (SNA-FL), seminaphthorhodafluors (SNARF), and BCE-CF using phase-modulation fluorometry. The phase and modulation values were found to be strongly pH-dependent in the physiological pH range, over the easily accessible range of light modulation frequencies from 10 to 300 MHz, making these ... More
Quantitative confocal imaging along the crypt-to-surface axis of colonic crypts.
AuthorsChu S, Brownell WE, Montrose MH
JournalAm J Physiol
PubMed ID8572186
Quantitative confocal microscopy methods are used to measure events along the crypt-to-surface axis in living mouse colonic mucosa. Experiments visualize carboxyseminaphthorhodofluor-1 (SNARF-1; a pH-sensitive fluorescent dye) in the extracellular fluid to measure extracellular pH within an intact epithelium. Lucifer yellow (LY; a pH-insensitive dye) is used to control for fidelity ... More
Monitoring intracellular pH changes in response to osmotic stress and membrane transport activity using 5-chloromethylfluorescein.
AuthorsSalvi A, Quillan JM, Sadée W
JournalAAPS PharmSci
PubMed ID12645993
Intracellular free H+ concentration (pHi) responds to numerous extracellular stimuli. The use of fluorescent indicator dyes to measure pHi is strongly influenced by the ability of target cells to retain activated dye within the cytoplasmic compartment. Here, 3 pH-sensitive indicator dye - acetoxymethyl (AM) esters of SNARF-1 and BCECF, and ... More
Intracellular pH measurement using single excitation-dual emission fluorescence ratios.
AuthorsBassnett S, Reinisch L, Beebe DC
JournalAm J Physiol
PubMed ID2301564
In the present paper, laser spectroscopy was used to evaluate the utility of a new fluorochrome, carboxyseminaphthorhodafluor-1 (Snarf-1), for single excitation-dual emission ratio measurement of intracellular pH (pHi). The emission spectrum of Snarf-1 showed clear pH-dependent shifts, and emission ratios calculated from the 640 and 587 nm maxima were a ... More
Characteristics of a store-operated calcium-permeable channel: sarcoendoplasmic reticulum calcium pump function controls channel gating.
AuthorsLiu X, Ambudkar IS
JournalJ Biol Chem
PubMed ID11395504
We examined the single channel properties and regulation of store-operated calcium channels (SOCC). In human submandibular gland cells, carbachol (CCh) induced flickery channel activity while thapsigargin (Tg) induced burst-like activity, with relatively lower open probability (NP(o)) and longer mean open time. Tg- and CCh-activated channels were permeable to Na(+) and ... More
Dopamine D2 receptor isoforms expressed in AtT20 cells differentially couple to G proteins to acutely inhibit high voltage-activated calcium channels.
AuthorsWolfe SE, Morris SJ
JournalJ Neurochem
PubMed ID10582596
The dopamine D2 receptor belongs to the serpentine superfamily of receptors, which have seven transmembrane segments and activate G proteins. D2 receptors are known to be linked, through Galpha(o)- and Galpha(i)-containing G proteins, to several signaling pathways in neuronal and secretory cells, including inhibition of adenylyl cyclase and high voltage-activated ... More
Proton and zinc effects on HERG currents.
AuthorsAnumonwo JM, Horta J, Delmar M, Taffet SM, Jalife J
JournalBiophys J
PubMed ID10388757
The proton and Zn2+ effects on the human ether-a-go-go related gene (HERG) channels were studied after expression in Xenopus oocytes and stable transfection in the mammalian L929 cell line. Experiments were carried out using the two-electrode voltage clamp at room temperature (oocytes) or the whole-cell patch clamp technique at 35 ... More
The contribution of pH-dependent mechanisms to fatigue at different intensities in mammalian single muscle fibres.
AuthorsChin ER, Allen DG
JournalJ Physiol
PubMed ID9769425
1. The contribution of intracellular pH (pHi) to the failure of Ca2+ release and inhibition of contractile proteins observed during fatigue was assessed in single intact mouse muscle fibres at 22 C. Fatigue was induced by repeated tetani at intensities designed to induce different levels of intracellular acidosis. Force and ... More
Determination of pH by microfluorometry: intracellular and interstitial pH regulation in developing early-stage fish embryos (Danio rerio).
AuthorsMölich A, Heisler N
JournalJ Exp Biol
PubMed ID16244172
Microfluorometric techniques were applied in vivo for continuous monitoring of specific acid-base parameters in zebrafish (Danio rerio) embryos during early stages of ontogeny. Dextran-coupled pH-sensitive single-excitation/dual-emission dye SNARF-1 was pressure-injected into individual cells or the interstitial space of 16- to 256-cell embryos, and pH was continuously recorded during subsequent development ... More
In the previous sections we demonstrated imaging of intracellular Ca2+ using our approach to FLIM. What other analytes can be imaged using FLIM? We have now characterized the lifetime of a good number of ion indicators. Based on these studies we know that Cl- can be imaged using FLIM with ... More
Calcium regulation of intracellular pH in pituitary intermediate lobe melanotropes.
AuthorsBeatty DM, Chronwall BM, Howard DE, Wiegmann TB, Morris SJ
JournalEndocrinology
PubMed ID8396024
The regulatory activities of both intracellular calcium ([Ca2+]i) and intracellular pH (pHi) have greatly increased interest in the study of their interdependence. We have designed an epifluorescence video microscope that will image the fluorescence from two ratio dyes, indo-1 (for [Ca2+]i) and SNARF-1 (for pHi) at video rates. We examined ... More
Evidence for redistribution-associated intracellular pK shifts of the pH-sensitive fluoroprobe carboxy-SNARF-1.
AuthorsOpitz N, Merten E, Acker H
JournalPflugers Arch
PubMed ID8072854
Properties and peculiarities of the pH-sensitive fluoroprobe carboxy-seminaphthorhodafluor-1 (carboxy-SNARF-1), in view of pHi measurements in single cells, were evaluated using confocal laser scanning microscopy. It was found that in human malignant glioma cells (U 118 MG) grown in multicellular spheroid culture, intracellular calibration curves (nigericin method) varied from one cell ... More
Spatiotemporal relationships among early events of fertilization in sea urchin eggs revealed by multiview microscopy.
AuthorsSuzuki K, Tanaka Y, Nakajima Y, Hirano K, Itoh H, Miyata H, Hayakawa T, Kinosita K
JournalBiophys J
PubMed ID7756541
Four early events of egg fertilization, changes in intracellular calcium concentration and intracellular pH, reorientation of the surface membrane, and the elevation of the fertilization envelope, were imaged in real time and in pairs in single sea urchin eggs. The paired imaging allowed the correlation of the four events spatially ... More
Lactate transport in mammalian ventricle. General properties and relation to K+ fluxes.
AuthorsShieh RC, Goldhaber JI, Stuart JS, Weiss JN
JournalCirc Res
PubMed ID8156630
Net cellular L-lactate efflux associated with accelerated anaerobic glycolysis has been implicated as a potential cause of the marked cellular K+ loss contributing to lethal cardiac arrhythmias in ischemic heart and to impaired function of fatigued skeletal muscle. To examine the mechanisms of transsarcolemmal L-lactate movement in the heart, isolated ... More
Targeting hypoxia regulated sodium driven bicarbonate transporters reduces triple negative breast cancer metastasis.
Authors
JournalNeoplasia
PubMed ID35150959
Molecular architecture underlying fluid absorption by the developing inner ear.
Authors
JournalElife
PubMed ID28994389
IL-12, IL-15, and IL-18 pre-activated NK cells target resistant T cell acute lymphoblastic leukemia and delay leukemia development in vivo.
Authors
JournalOncoimmunology
PubMed ID28405496
Root vacuolar sequestration and suberization are prominent responses of Pistacia spp. rootstocks during salinity stress.
Authors
JournalPlant Direct
PubMed ID34027297
Iron-deficiency anemia reduces cardiac contraction by downregulating RyR2 channels and suppressing SERCA pump activity.