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
CFTR regulates phagosome acidification in macrophages and alters bactericidal activity.
AuthorsDi A, Brown ME, Deriy LV, Li C, Szeto FL, Chen Y, Huang P, Tong J, Naren AP, Bindokas V, Palfrey HC, Nelson DJ
JournalNat Cell Biol
PubMed ID16921366
'Acidification of phagosomes has been proposed to have a key role in the microbicidal function of phagocytes. Here, we show that in alveolar macrophages the cystic fibrosis transmembrane conductance regulator Cl- channel (CFTR) participates in phagosomal pH control and has bacterial killing capacity. Alveolar macrophages from Cftr-/- mice retained the ... More
Rapid flow cytometric method for measuring senescence associated beta-galactosidase activity in human fibroblasts.
AuthorsNoppe G, Dekker P, de Koning-Treurniet C, Blom J, van Heemst D, Dirks RW, Tanke HJ, Westendorp RG, Maier AB,
JournalCytometry A
PubMed ID19777541
'Senescence associated-beta-galactosidase (SA-beta-gal) activity is a widely used marker for cellular senenescence. SA-beta-gal activity is routinely detected cytochemically, manually discriminating negative from positive cells. This method is time-consuming, subjective and therefore prone to operator-error. We aimed to optimize a flow cytometric method described by other workers using endothelial cells to ... More
Effects of cytoplasmic acidification on clathrin lattice morphology.
AuthorsHeuser J
JournalJ Cell Biol
PubMed ID2563729
'Reducing the internal pH of cultured cells by several different protocols that block endocytosis is found to alter the structure of clathrin lattices on the inside of the plasma membrane. Lattices curve inward until they become almost spherical yet remain stubbornly attached to the membrane. Also, the lattices bloom empty ... More
Estimation of intracellular chloride activity in isolated perfused rabbit proximal convoluted tubules using a fluorescent indicator.
AuthorsKrapf R, Berry CA, Verkman AS
JournalBiophys J
PubMed ID3395662
'The methodology has been developed to measure cell chloride activity by fluorescence microscopy using the chloride-sensitive dye, 6-methoxy-1-(3-sulfonatopropyl)quinolinium (SPQ). SPQ was loaded into cells of the in vitro microperfused rabbit proximal convoluted tubule by a 10 min luminal perfusion with 20 mM SPQ at 38 degrees C. Fluorescence was excited ... More
Ca2+ storage in Trypanosoma brucei: the influence of cytoplasmic pH and importance of vacuolar acidity.
AuthorsScott DA, Moreno SN, Docampo R
JournalBiochem J
PubMed ID7575411
'The hypothesis that changes in cytosolic pH effect the release from intracellular compartments of stored calcium in Trypanosoma brucei was addressed by the use of procyclic and bloodstream trypomastigotes of T. brucei loaded with the fluorescent reagents 2'',7''-bis-(2-carboxyethyl)-5(and 6)-carboxyfluorescein (BCECF) to measure intracellular pH (pHi), or fura 2 to measure ... More
Determination of the intracellular free chloride concentration in rat brain synaptoneurosomes using a chloride-sensitive fluorescent indicator.
AuthorsEngblom AC, Akerman KE
JournalBiochim Biophys Acta
PubMed ID8274496
'The chloride-sensitive fluorescent indicator MQAE (N-(6-methoxyquinolyl) acetoacetyl ester) has been used for determination of the intracellular free chloride concentration in rat brain synaptoneurosomes. Loading of the synaptoneurosomes with MQAE occurs by transmembrane diffusion. Calibration of the intracellular MQAE was done by determining the correlation between fluorescence intensity and intrasynaptoneurosomal Cl- ... More
Regulation of intracellular pH in single rat zona glomerulosa cells.
AuthorsShepherd RM, Williams GH, Quinn SJ
JournalAm J Physiol
PubMed ID1310209
'The cytosolic pH (pHi) regulation of rat adrenal zona glomerulosa (ZG) cells was studied using single-cell spectrofluorimetry. Basal pHi was similar for cells incubated in the absence or presence of the HCO3(-)-CO2 buffering system. In the absence of HCO3-, inhibition of the Na(+)-H+ exchanger by dimethylamiloride (DMA) or removal of ... More
Epidermal growth factor elevates intracellular pH in chicken granulosa cells.
AuthorsLi M, Morley P, Asem EK, Tsang BK
JournalEndocrinology
PubMed ID1649743
'Many bioregulators, such as epidermal growth factor (EGF), induce intracellular alkalinization by activating a membrane bound Na+/H+ antiporter. The present studies were designed to examine the influence of EGF on intracellular pH (pHi) in chicken granulosa cells. pHi in granulosa cells from the two largest preovulatory follicles of hens was ... More
A new action of parathyroid hormone. receptor-mediated stimulation of extracellular acidification in human osteoblast-like SaOS-2 cells.
AuthorsBarrett MG, Belinsky GS, Tashjian AH
JournalJ Biol Chem
PubMed ID9334207
'The major physiological function of parathyroid hormone (PTH) is the maintenance of Ca2+/Pi homeostasis via the parathyroid hormone/parathyroid hormone-related protein receptor (PTHR) in kidney and bone. An important consequence of PTHR activation in bone is enhanced local acidification of the extracellular space. Agonist activation of some seven transmembrane-domain receptors increases ... More
Effects of osmotic stresses on isolated rat hepatocytes. II. Modulation of intracellular pH.
AuthorsGleeson D, Corasanti JG, Boyer JL
JournalAm J Physiol
PubMed ID2305896
'To assess the roles of acid-base transport systems in cell volume regulation in rat hepatocytes, intracellular pH (pHi) was measured in subconfluent monolayers loaded with 2''-7''-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF) after exposure to hypotonic and relative hypertonic media, interventions that stimulate regulatory volume decrease (RVD) and increase (RVI), respectively. During RVD, pHi decreased ... More
Actin assembly in electropermeabilized neutrophils: role of intracellular calcium.
AuthorsDowney GP, Chan CK, Trudel S, Grinstein S
JournalJ Cell Biol
PubMed ID2112547
'Assembly of microfilaments involves the conversion of actin from the monomeric (G) to the filamentous (F) form. The exact sequence of events responsible for this conversion is yet to be defined and, in particular, the role of calcium remains unclear. Intact and electropermeabilized human neutrophils were used to assess more ... More
Suppression of Ca2+ oscillations in cultured rat hepatocytes by chemical hypoxia.
AuthorsKawanishi T, Nieminen AL, Herman B, Lemasters JJ
JournalJ Biol Chem
PubMed ID1939069
'The model of "chemical hypoxia" with KCN plus iodoacetic acid mimics the ATP depletion and reductive stress of hypoxia. Here, we examined the effects of chemical hypoxia on cytosolic free Na+ and Ca2+ in single cultured rat hepatocytes by multiparameter digitized video microscopy and ratio imaging of sodium-binding furan indicator ... More
Acetate transport in the S3 segment of the rabbit proximal tubule and its effect on intracellular pH.
AuthorsNakhoul NL, Boron WF
JournalJ Gen Physiol
PubMed ID3225555
'We monitored intracellular pH (pHi) in isolated perfused S3 segments of the rabbit proximal tubule, and studied the effect of acetate (Ac-) transport on pHi. pHi was calculated from the absorbance spectrum of 4',5'-dimethyl-5-(and 6) carboxyfluorescein trapped intracellularly. All solutions were nominally HCO3(-)-free. Removal of 10 mM Ac- from bath ... More
Acidification of the cytosol inhibits endocytosis from coated pits.
AuthorsSandvig K, Olsnes S, Petersen OW, van Deurs B
JournalJ Cell Biol
PubMed ID2887575
'Acidification of the cytosol of a number of different cell lines strongly reduced the endocytic uptake of transferrin and epidermal growth factor. The number of transferrin binding sites at the cell surface was increased in acidified cells. Electron microscopic studies showed that the number of coated pits at the cell ... More
Estimation of matrix pH in isolated heart mitochondria using a fluorescent probe.
AuthorsJung DW, Davis MH, Brierley GP
JournalAnal Biochem
PubMed ID2751096
'Isolated heart mitochondria hydrolyze the acetoxymethyl esters of the Ca2+-sensitive fluorescent probe fura-2 and the pH-sensitive 2'',7''-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF). The resulting charged forms of the probes are retained in the mitochondrial matrix and appear well-suited for the estimation of pCa and pH in this compartment. The mitochondria esterase activity is stimulated ... More
Effect of heat shock, [Ca2+]i, and cAMP on inositol trisphosphate in human epidermoid A-431 cells.
AuthorsKiang JG, McClain DE
JournalAm J Physiol
PubMed ID8392802
'The basal levels of inositol monophosphate, inositol bisphosphate, and inositol trisphosphate (InsP3) in A-431 cells incubated in Na(+)-Hanks'' solution were, respectively, 1.23 +/- 0.18, 0.17 +/- 0.03, and 0.69 +/- 0.07% of the total radioactivity in the cell. When cells were heated, InsP3 increased in a temperature-dependent manner related to ... More
Calcium binding to the cytosol and calcium extrusion mechanisms in intact synaptosomes and their alterations with aging.
AuthorsMartinez-Serrano A, Blanco P, Satrústegui J
JournalJ Biol Chem
PubMed ID1531657
'A simple method to measure cytosolic calcium binding in intact presynaptic nerve terminals (synaptosomes) from rat brain, which is based on the simultaneous determination of [Ca2+]i and total [45Ca2+] in quin2-loaded synaptosomes undergoing a switch from high- to low-calcium containing medium, is presented. Binding to the cytosolic compartment alone was ... More
Identification of residues lining the translocation pore of human AE1, plasma membrane anion exchange protein.
AuthorsTang XB, Kovacs M, Sterling D, Casey JR
JournalJ Biol Chem
PubMed ID9920902
'AE1 is the chloride/bicarbonate anion exchanger of the erythrocyte plasma membrane. We have used scanning cysteine mutagenesis and sulfhydryl-specific chemistry to identify pore-lining residues in the Ser643-Ser690 region of the protein. The Ser643-Ser690 region spans transmembrane segment 8 of AE1 and surrounds Glu681, which may reside at the transmembrane permeability ... More
Continuous measurement of mitochondrial pH gradients in isolated hepatocytes by difference ratio spectroscopy.
AuthorsThomas PJ, Gaspers LD, Pharr C, Thomas JA
JournalArch Biochem Biophys
PubMed ID1898020
'The pH gradient, delta pH, present across the inner mitochondrial membrane in isolated rat hepatocytes was continuously monitored with a novel spectroscopic technique that utilizes the weak acid fluorescein. Unlike most cytosolic pH indicators, such as 2'',7''-bis(carboxyethyl)-5,(6)-carboxyfluorescein (BCECF), fluorescein freely distributes between the cytosolic and mitochondrial compartments. As is typical ... More
An alkaline pH-activated Cl(-)-anion exchanger regulates pH homeostasis in fibroblasts.
AuthorsLin PY, Ahluwalia M, Gruenstein E
JournalAm J Physiol
PubMed ID1689112
'Alkalinization of the cytoplasm in human fibroblasts activates Cl- efflux. The pathway of Cl- efflux is inhibited by 4,4''-diisothiocyanostilbene-2,2''-disulfonic acid (DIDS) or by removal of Cl- from the extracellular medium, but it is insensitive to the Cl(-)-cation cotransport inhibitor bumetanide. These data indicate that the alkali-stimulated Cl- pathway is a ... More
Dynamical change of mitochondrial DNA induced in the living cell by perturbing the electrochemical gradient.
AuthorsCoppey-Moisan M, Brunet AC, Morais R, Coppey J
JournalBiophys J
PubMed ID8913573
'Digital-imaging microscopy was used in conditions that allowed the native state to be preserved and hence fluorescence variations of specific probes to be followed in the real time of living mammalian cells. Ethidium bromide was shown to enter into living cells and to intercalate stably into mitochondrial DNA (mtDNA), giving ... More
The effects of metabolic inhibition on intracellular calcium and pH in isolated rat ventricular cells.
AuthorsEisner DA, Nichols CG, O'Neill SC, Smith GL, Valdeolmillos M
JournalJ Physiol
PubMed ID2614727
'1. Intracellular calcium concentration [( Ca2+]i) and pH (pHi) were measured in single, isolated rat ventricular myocytes using, respectively, the fluorescent indicators Fura-2 and BCECF (2',7'-bis(carboxyethyl-5(6)-carboxyfluorescein). Contraction was measured simultaneously. The intracellular calibration of BCECF is demonstrated. In a HEPES-buffered bathing solution of pH 7.4, pHi had a mean value ... More
Mutations in the P. falciparum digestive vacuole transmembrane protein PfCRT and evidence for their role in chloroquine resistance.
AuthorsFidock DA, Nomura T, Talley AK, Cooper RA, Dzekunov SM, Ferdig MT, Ursos LM, Sidhu AB, Naudé B, Deitsch KW, Su XZ, Wootton JC, Roepe PD, Wellems TE
JournalMol Cell
PubMed ID11090624
'The determinant of verapamil-reversible chloroquine resistance (CQR) in a Plasmodium falciparum genetic cross maps to a 36 kb segment of chromosome 7. This segment harbors a 13-exon gene, pfcrt, having point mutations that associate completely with CQR in parasite lines from Asia, Africa, and South America. These data, transfection results, ... More
Influence of acid secretory state on Cl(-)-base and Na(+)-H+ exchange and pHi in isolated rabbit parietal cells.
AuthorsSeidler U, Roithmaier S, Classen M, Silen W
JournalAm J Physiol
PubMed ID1310223
'Parietal cell apical proton secretion is accompanied by apical Cl- secretion, K+ cycling, and the generation of intracellular base. We examined the ability of the parietal cell to maintain intracellular pH (pHi) during its transformation from the resting to the stimulated state and evaluated the ion transport mechanisms involved in ... More
The Na+/H+ antiporter in rat alveolar type II cells and its role in stimulated surfactant secretion.
AuthorsSano K, Cott GR, Voelker DR, Mason RJ
JournalBiochim Biophys Acta
PubMed ID2833308
'We used the pH-sensitive fluorescent probe 2'',7''-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) to identify Na+/H+ exchange in freshly isolated rat alveolar type II cells and alveolar type II cells in primary culture. The intracellular pH (pHi) of freshly isolated alveolar type II cells was 7.36 +/- 0.05 (n = 3). When freshly isolated alveolar ... 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
Transporters involved in regulation of intracellular pH in primary cultured rat brain endothelial cells.
AuthorsTaylor CJ, Nicola PA, Wang S, Barrand MA, Hladky SB,
JournalJ Physiol
PubMed ID16916905
'Fluid secretion across the blood-brain barrier, critical for maintaining the correct fluid balance in the brain, entails net secretion of HCO(3)(-), which is brought about by the combined activities of ion transporters situated in brain microvessels. These same transporters will concomitantly influence intracellular pH (pH(i)). To analyse the transporters that ... More
Disease-causing mutations in the cystic fibrosis transmembrane conductance regulator determine the functional responses of alveolar macrophages.
'Alveolar macrophages (AMs) play a major role in host defense against microbial infections in the lung. To perform this function, these cells must ingest and destroy pathogens, generally in phagosomes, as well as secrete a number of products that signal other immune cells to respond. Recently, we demonstrated that murine ... More
Characterization of the activation of Na+/H+ exchange in lymphocytes by phorbol esters: change in cytoplasmic pH dependence of the antiport.
AuthorsGrinstein S, Cohen S, Goetz JD, Rothstein A, Gelfand EW
JournalProc Natl Acad Sci U S A
PubMed ID2983345
'Phorbol 12-myristate 13-acetate and other phorbol esters induce an intracellular alkalinization in rat thymic lymphocytes. An extracellular acidification can be recorded concomitantly. This transmembrane H+ (equivalent) flux is dependent on external Na+ and is amiloride sensitive. Phorbol esters also stimulate an amiloride-sensitive uptake of 22Na+, suggesting activation of Na+o/H+i exchange. ... More
A role for mitochondria in NLRP3 inflammasome activation.
AuthorsZhou R, Yazdi AS, Menu P, Tschopp J,
JournalNature
PubMed ID21124315
'An inflammatory response initiated by the NLRP3 inflammasome is triggered by a variety of situations of host ''danger'', including infection and metabolic dysregulation. Previous studies suggested that NLRP3 inflammasome activity is negatively regulated by autophagy and positively regulated by reactive oxygen species (ROS) derived from an uncharacterized organelle. Here we ... More
Interrelationship between growth factor-induced pH changes and intracellular Ca2+.
AuthorsIves HE, Daniel TO
JournalProc Natl Acad Sci U S A
PubMed ID3470769
'Many mitogens cause rapid changes in intracellular pH and Ca2+. We studied the patterns of pH and Ca2+ changes after exposure of murine fibroblasts to platelet-derived growth factor (PDGF), bombesin, phorbol 12-myristate 13-acetate (PMA), and the vasoactive peptide bradykinin. Intracellular pH and Ca2+ were measured by using the fluorescent dyes ... More
Mechanisms of mitochondrial photosensitization by the cationic dye, N,N-bis(2-ethyl-1,3-dioxylene)kryptocyanine (EDKC): preferential inactivation of complex I in the electron transport chain.
AuthorsAra G, Aprille JR, Malis CD, Kane SB, Cincotta L, Foley J, Bonventre JV, Oseroff AR
JournalCancer Res
PubMed ID3119197
'We investigated mechanisms of mitochondrial phototoxicity caused by the cationic cyanine dye N,N''-bis(2-ethyl-1,3-dioxylene)kryptocyanine (EDKC), examining the role of the mitochondrial membrane potential on the dye uptake by carcinoma cells in vitro, and both the dark and photosensitizing effects of the dye on the function of isolated mouse liver mitochondria. When ... More
Photoactivated azido fatty acid irreversibly inhibits anion and proton transport through the mitochondrial uncoupling protein.
AuthorsJezek P, Hanus J, Semrad C, Garlid KD
JournalJ Biol Chem
PubMed ID8626410
'The protonophoretic function of uncoupling protein (UCP) is activated by fatty acids. According to the "docking site" hypothesis (Jezek, P., and Garlid, K. D., J. Biol. Chem. 265, 19303-19311, 1990), the fatty acid binding site is identical with the anion channel of UCP. Skulachev (Skulachev, V. P. (1991) FEBS Lett. ... More
Na+-H+ and Cl(-)-OH-(HCO3-) exchange in gastric glands.
AuthorsParadiso AM, Negulescu PA, Machen TE
JournalAm J Physiol
PubMed ID3008575
'The pH-sensitive, fluorescent, cytoplasmic-trapped dye 2,7-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) has been used to measure intracellular (pHi) and pH electrode to measure extracellular pH (pHo) in suspensions of gastric glands isolated from rabbit stomachs. The fluorescence of BCECF-loaded glands was calibrated in terms of pHi by equilibrating pHo and pHi using ionophores or ... More
Serum regulates Na+/H+ exchange in Caco-2 cells by a mechanism which is dependent on F-actin.
AuthorsWatson AJ, Levine S, Donowitz M, Montrose MH
JournalJ Biol Chem
PubMed ID1309780
'Regulation of Na+/H+ exchange by fetal bovine serum was studied in Caco-2 cells, an established cell line derived from a human colon carcinoma. Cells were grown as polarized monolayers on collagen-coated filters and intracellular pH measured fluorometrically with 2'',7''-bis(2-carboxymethyl)-5,6-carboxyfluorescein. Na+/H+ exchange was reduced 64% when cells were deprived of serum ... More
Mitochondrial K+ as modulator of Ca(2+)-dependent cytotoxicity in hepatocytes. Novel application of the K(+)-sensitive dye PBFI (K(+)-binding benzofuran isophthalate) to assess free mitochondrial K+ concentrations.
AuthorsZoeteweij JP, van de Water B, de Bont HJ, Nagelkerke JF
JournalBiochem J
PubMed ID8172616
'In isolated rat hepatocytes a sustained high increase in intracellular free Ca2+ ([Ca2+]i), induced by extracellular ATP, is associated with mitochondrial dysfunction and cell death. The Ca(2+)-induced effects are Pi-dependent and less severe when the intracellular K+ content is low. In this study, the involvement of mitochondrial K+ processing in ... More
Regulation of the mitochondrial Na+/Ca2+ antiport by matrix pH.
'The effect of matrix pH (pHi) on the activity of the mitochondrial Na+/Ca2+ antiport has been studied using the fluorescence of SNARF-1 to monitor pHi and Na(+)-dependent efflux of accumulated Ca2+ to follow antiport activity. Heart mitochondria respiring in a KCl medium maintain a large delta pH (interior alkaline) and ... More
Secretagogue-induced changes in intracellular pH and amylase release in mouse pancreatic acini.
AuthorsCarter KJ, Rutledge PL, Steer ML, Silen W
JournalAm J Physiol
PubMed ID2446509
'The response of the intracellular pH (pHi) to stimulation of enzyme secretion in pancreatic acini was measured using the fluorescent dye 2''-7''-bis(carboxyethyl)-5(6)-carboxyfluorescein. Acini suspended in pH 7.40 buffer demonstrated cytoplasmic alkalinization of 0.17, 0.14, and 0.15 pH units 2 min after addition of the secretagogues carbachol (10(-5) M), caerulein (10(-10) ... More
Proline-rich motifs of the Na+/H+ exchanger 2 isoform. Binding of Src homology domain 3 and role in apical targeting in epithelia.
AuthorsChow CW, Woodside M, Demaurex N, Yu FH, Plant P, Rotin D, Grinstein S, Orlowski J
JournalJ Biol Chem
PubMed ID10187839
'The NHE2 isoform of the Na+/H+ exchanger (NHE) displays two proline-rich sequences in its C-terminal region that resemble SH3 (Src homology 3)-binding domains. We investigated whether these regions (743PPSVTPAP750, termed Pro-1, and 786VPPKPPP792, termed Pro-2) can bind to SH3 domains and whether they are essential for NHE2 function and targeting. ... More
Regulation of intracellular pH in rat lactotrophs: involvement of anionic exchangers.
AuthorsGarcia L, Boué-Grabot E, Garret M, Sartor P
JournalEndocrinology
PubMed ID9322929
'Regulation of the intracellular pH (pHi) of normal rat lactotrophs was studied. As this cell type, cultured with 10% FCS, can achieve a relatively alkaline pHi (7.3-7.5), we investigated the presence of a mechanism based on Cl-/HCO3- exchange. Using the pHi-sensitive probe SNARF-1 (seminaphtorodafluor) in its permeant form, SNARF-1/AM, we ... More
The Na/K/2Cl cotransporter is increased in hypertrophied vascular smooth muscle cells.
AuthorsTseng H, Berk BC
JournalJ Biol Chem
PubMed ID1569072
'Hypertrophy of vascular smooth muscle cells (VSMC) is a pathogenic feature of hypertension which may contribute to abnormal vessel tone and function. As a consequence of the increase in cell size associated with hypertrophy, it is likely that alterations in the mechanisms that regulate VSMC intracellular volume occur. Because the ... More
The lysosomal proton pump is electrogenic.
AuthorsHarikumar P, Reeves JP
JournalJ Biol Chem
PubMed ID6224789
'Lysosomes were purified approximately 40-fold from rat kidney cortex by differential and Percoll density gradient centrifugation. In a sucrose medium, the lysosomes quenched the fluorescence of the potential sensitive dye diS-C3-(5) (3,3''-dipropylthiocarbo-cyanine iodide) in a time-dependent manner, indicating that the dye accumulates within the lysosomal interior. After treatment of the ... More
Bioenergetics of the staphylococcal multidrug export protein QacA. Identification of distinct binding sites for monovalent and divalent cations.
AuthorsMitchell BA, Paulsen IT, Brown MH, Skurray RA
JournalJ Biol Chem
PubMed ID9920900
'The multidrug efflux pump QacA from Staphylococcus aureus confers resistance to an extensive range of structurally dissimilar compounds. Fluorimetric analyses demonstrated that QacA confers resistance to the divalent cation 4'',6-diamidino-2-phenylindole, utilizing a proton motive force-dependent efflux mechanism previously demonstrated for QacA-mediated resistance to the monovalent cation ethidium. Both the ionophores ... More
Na(+)-H+ and Na(+)-dependent Cl(-)-HCO3- exchange control pHi in vascular smooth muscle.
AuthorsKahn AM, Cragoe EJ, Allen JC, Halligan RD, Shelat H
JournalAm J Physiol
PubMed ID2164779
'The mechanisms that control intracellular pH (pHi) in vascular smooth muscle are not fully understood. These studies were performed to determine the identity and relative importance of the sarcolemmal transport systems that mediate net acid efflux in primary cultured vascular smooth muscle cells from canine femoral artery. In HEPES- or ... More
Preparation of giant liposomes in physiological conditions and their characterization under an optical microscope.
AuthorsAkashi K, Miyata H, Itoh H, Kinosita K
JournalBiophys J
PubMed ID8968594
'Unilamellar liposomes with diameters of 25-100 microns were prepared in various physiological salt solutions, e.g., 100 mM KCl plus 1 mM CaCl2. Successful preparation of the giant liposomes at high ionic strengths required the inclusion of 10-20% of a charged lipid, such as phosphatidylglycerol, phosphatidylserine, phosphatidic acid, or cardiolipin, in ... More
Trapping of inhibitor-induced conformational changes in the erythrocyte membrane anion exchanger AE1.
AuthorsTang XB, Casey JR
JournalBiochemistry
PubMed ID10545179
'AE1, the chloride/bicarbonate anion exchanger of the erythrocyte plasma membrane, is highly sensitive to inhibition by stilbene disulfonate compounds such as DIDS (4,4''-diisothiocyanostilbene-2, 2''-disulfonate) and DNDS (4,4''-dinitrostilbene-2,2''-disulfonate). Stilbene disulfonates recruit the anion binding site to an outward-facing conformation. We sought to identify the regions of AE1 that undergo conformational changes ... More
Evidence that Na+/H+ exchange regulates angiotensin II-stimulated diacylglycerol accumulation in vascular smooth muscle cells.
AuthorsGriendling KK, Berk BC, Alexander RW
JournalJ Biol Chem
PubMed ID2839498
'Angiotensin II stimulation of vascular smooth muscle cells results in initial, rapid diacylglycerol (DG) formation from the polyphosphoinositides accompanied by intracellular acidification, as well as a more sustained DG accumulation which is accompanied by a prolonged intracellular alkalinization. To determine whether intracellular pH (pHi) modulates DG accumulation, NH4Cl and potassium ... More
Confocal imaging of intracellular chloride in living brain slices: measurement of GABAA receptor activity.
AuthorsInglefield JR, Schwartz-Bloom RD
JournalJ Neurosci Methods
PubMed ID9288644
'We have developed a method using UV laser-scanning confocal microscopy and the fluorescent chloride ion indicator, 6-methoxy-N-ethylquinolinium chloride (MEQ), to image GABA-mediated changes in intracellular chloride (Cli-) in individual neurons of the rat acute brain slice. After bath-loading slices with the cell-permeant form (reduced) of MEQ, there was intense fluorescence ... More
Intracellular Ca2+ concentration and pHi during metabolic inhibition.
AuthorsHayashi H, Miyata H, Noda N, Kobayashi A, Hirano M, Kawai T, Yamazaki N
JournalAm J Physiol
PubMed ID1550207
'To study the changes in intracellular Ca2+ concentration ([Ca2+]i) and pH (pHi) during metabolic inhibition, rat ventricular myocytes were dual loaded with the acetoxymethyl esters of fura-2 (fura-2/AM) and 2'',7''-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF/AM) and perfused with 2 mM sodium cyanide (NaCN). The percent of rod-shaped cells was 30% of the control after ... More
Simultaneous measurement of intracellular pH and Ca2+ using the fluorescence of SNARF-1 and fura-2.
'Upon cell stimulation with hormones and other mitogens, a variety of biochemical and physiological responses occur within the first few minutes. Changes in both intracellular pH (pHin) and intracellular Ca2+ concentration ([Ca2+]in) are prominent and play a major role in the signal transduction mechanism leading to the physiological response, i.e., ... More
Cl- fluxes related to fluid secretion by the rat parotid: involvement of Cl(-)-HCO3- exchange.
AuthorsMelvin JE, Turner RJ
JournalAm J Physiol
PubMed ID1312791
'Muscarinic-induced 36Cl- and 86Rb+ (K+ substitute) fluxes were studied in rat parotid acini. Stimulation resulted in a rapid [half time (t1/2) less than 30 s] decrease in both Cl- and Rb+ content (approximately 50 and 30%, respectively) followed by a slower partial recovery (t1/2 approximately 3-4 min) to approximately 80% ... More
Flow cytometric measurement of [Ca2+]i and pHi in conjugated natural killer cells and K562 target cells during the cytotoxic process.
AuthorsVan Graft M, Kraan YM, Segers IM, Radosevic K, De Grooth BG, Greve J
JournalCytometry
PubMed ID8472603
'We describe a flow cytometric assay that enables one to follow conjugate formation between cytotoxic cells and their target cells during the cytotoxic process. In addition, the internal calcium concentration ([Ca2+]i) and internal pH (pHi) of the conjugated cells can be monitored and directly compared to the nonconjugated cells. This ... More
Intracellular pH regulation by Na(+)/H(+) exchange requires phosphatidylinositol 4,5-bisphosphate.
AuthorsAharonovitz O, Zaun HC, Balla T, York JD, Orlowski J, Grinstein S
JournalJ Cell Biol
PubMed ID10893269
'The carrier-mediated, electroneutral exchange of Na(+) for H(+) across the plasma membrane does not directly consume metabolic energy. Nevertheless, acute depletion of cellular ATP markedly decreases transport. We analyzed the possible involvement of polyphosphoinositides in the metabolic regulation of NHE1, the ubiquitous isoform of the Na(+)/H(+) exchanger. Depletion of ATP ... More
Biochemical properties of vacuolar zinc transport systems of Saccharomyces cerevisiae.
AuthorsMacDiarmid CW, Milanick MA, Eide DJ
JournalJ Biol Chem
PubMed ID12161436
'The yeast vacuole plays an important role in zinc homeostasis by storing zinc for later use under deficient conditions, sequestering excess zinc for its detoxification, and buffering rapid changes in intracellular zinc levels. The mechanisms involved in vacuolar zinc sequestration are only poorly characterized. Here we describe the properties of ... More
Human neutrophil stimulation by influenza virus: relationship of cytoplasmic pH changes to cell activation.
AuthorsHartshorn KL, Wright J, Collamer MA, White MR, Tauber AI
JournalAm J Physiol
PubMed ID2113768
'We have previously demonstrated that influenza A virus (IAV) stimulates the human neutrophil through phospholipase C activation. With the use of the fluorescent indicator 2'',7''-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF), cytoplasmic acidification and subsequent alkalinization are shown to accompany this activation. These responses are not inhibited by pertussis toxin (PT). The alkalinization is mediated ... More
A novel method for absolute calibration of intracellular pH indicators.
AuthorsEisner DA, Kenning NA, O'Neill SC, Pocock G, Richards CD, Valdeolmillos M
JournalPflugers Arch
PubMed ID2740208
'In this paper we present methods to measure intracellular pH (pHi) with fluorescent indicators. These methods are based on the change in intracellular pH following the addition of weak acids and weak bases to the extracellular medium. The first method requires that the fluorescence of the indicator is proportional to ... More
Thallium mediates a rapid chloride/hydroxyl ion exchange through myelin lipid bilayers.
AuthorsDíaz RS, Monreal J
JournalMol Pharmacol
PubMed ID7808444
'We have investigated the effects of several heavy metal cations on the proton and chloride permeabilities of liposomes prepared with endogenous lipids from brain myelin, by monitoring the fluorescence emitted by acridine orange and N-(6-methoxyquinolyl)acetoethyl ester. In addition to Hg2+ and Cu+, nanomolar concentrations of Tl3+, but not Tl+, were ... More
Cytoplasmic [Ca2+] and intracellular pH in lymphocytes. Role of membrane potential and volume-activated Na+/H+ exchange.
AuthorsGrinstein S, Cohen S
JournalJ Gen Physiol
PubMed ID3559512
'The effect of elevating cytoplasmic Ca2+ [( Ca2+]i) on the intracellular pH (pHi) of thymic lymphocytes was investigated. In Na+-containing media, treatment of the cells with ionomycin, a divalent cation ionophore, induced a moderate cytoplasmic alkalinization. In the presence of amiloride or in Na+-free media, an acidification was observed. This ... More
Herbimycin A inhibits both dephosphorylation and translocation of cofilin induced by opsonized zymosan in macrophagelike U937 cells.
'We previously reported that a 21-kDa phosphoprotein may play an important role in superoxide production through dephosphorylation by neutrophillike differentiated HL-60 cells (Suzuki et al., 1995, Biochim Biophys Acta 1266: 261-267). The phosphoprotein was identified as cofilin, an actin-binding protein, and the activation-induced changes in its intracellular distribution have been ... More
Intracellular pH regulation in basal corneal epithelial cells measured in corneal explants: characterization of Na/H exchange.
AuthorsBonanno JA, Machen TE
JournalExp Eye Res
PubMed ID2547642
'Intracellular pH (pHi) was measured in basal corneal epithelial cells from fresh corneal explants using the pH sensitive fluorescent dye 2'',7''-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF). The overlying superficial and wing cells were removed by mechanical scraping to expose basal cells attached to their basal lamina. Tissue pieces with attached, dye-loaded basal cells were ... More
Cellular pH and Na+-H+ exchange activity in lens epithelium of Bufo marinus toad.
AuthorsWolosin JM, Alvarez LJ, Candia OA
JournalAm J Physiol
PubMed ID2847535
'Incubation of lenses with the acetoxymethyl ester of 2,7-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF-AM) led to selective, prolonged entrapment of the deesterified, pH-sensitive, membrane-impermeable form of the dye (BCECF) in the lens epithelial cells allowing, thereby, for the continuous fluorometric monitoring of epithelial intracellular pH (pHi) in intact lenses. Under anaerobic conditions in physiological ... More
Role of calcium and cAMP messenger systems in intracellular pH regulation of osteoblastic cells.
AuthorsGreen J, Kleeman CR
JournalAm J Physiol
PubMed ID1310204
'We have recently shown that two mechanisms are involved in the regulation of pHi in the osteoblastic phenotype cell line UMR-106 (Na(+)-H+ antiporter and a Na(+)-independent Cl(-)-HCO 3(-)-OH- exchanger). In the present work, we used the pH-sensitive dye 2'',7''-bis(2-carboxyethyl)-5,6-carboxyfluorescein as well as isotope fluxes to investigate the influence of second ... More
Characterization of sustained [Ca2+]i increase in pancreatic acinar cells and its relation to amylase secretion.
AuthorsTsunoda Y, Stuenkel EL, Williams JA
JournalAm J Physiol
PubMed ID1700626
'The sustained increase in cytosolic free Ca2+ concentration ([Ca2+]i) during maximal stimulation of rat pancreatic acini with carbamylcholine (10(-5) M) was investigated in individual acinar cells by microspectrofluorometric analysis of fura-2. After the large initial [Ca2+]i increase from intracellular stores, [Ca2+]i remained significantly elevated as long as the stimulus was ... More
Leucine transport-induced activation of the Na+/H+ exchanger in human peripheral lymphocytes.
AuthorsMitsumoto Y, Sato K, Mohri T
JournalBiochim Biophys Acta
PubMed ID2833306
'Adjustment of amino-acid-induced cytoplasmic pH decrease by the Na+/H+ exchange system in human lymphocytes has been studied using a fluorometric technique to monitor the intracellular pH change. When the interior of lymphocytes is acidified by addition of nigericin to medium, cytoplasmic pH is immediately corrected toward its resting value. This ... More
Intracellular alkalinization leads to Ca2+ mobilization from agonist-sensitive pools in bovine aortic endothelial cells.
AuthorsDanthuluri NR, Kim D, Brock TA
JournalJ Biol Chem
PubMed ID2172243
'Receptor-stimulated phosphoinositide turnover leads to activation of Na+/H+ exchange and subsequent intracellular alkalinization. To probe the effect of increased intracellular pH (pHi) on Ca2+ homeostasis in cultured bovine aortic endothelial cells (BAEC), we studied the effect of weak bases, ammonium chloride (NH4Cl) and methylamine (agents which increase pHi by direct ... More
Relationship of the effects of nigericin on the aggregation and cytoplasmic pH of bovine platelets in the presence of different cations.
AuthorsKitagawa S, Awai M, Kametani F
JournalBiochim Biophys Acta
PubMed ID2441757
'The effect of nigericin on aggregation of bovine platelets was investigated in media containing the chloride salts of various alkali metal cations of quaternary ammonium cations. In medium with K+, which has the highest permeability with the ionophore among the cations tested, nigericin slightly enhanced both ADP- and thrombin-induced aggregation. ... More
Cytosolic pH regulation in osteoblasts. Regulation of anion exchange by intracellular pH and Ca2+ ions.
AuthorsGreen J, Yamaguchi DT, Kleeman CR, Muallem S
JournalJ Gen Physiol
PubMed ID2299328
'Measurements of cytosolic pH (pHi) 36Cl fluxes and free cytosolic Ca2+ concentration ([Ca2+]i) were performed in the clonal osteosarcoma cell line UMR-106 to characterize the kinetic properties of Cl-/HCO3- (OH-) exchange and its regulation by pHi and [Ca2+]i. Suspending cells in Cl(-)-free medium resulted in rapid cytosolic alkalinization from pHi ... More
Effect of cytoplasmic pH on platelet activation by arachidonic acid.
AuthorsIshikawa Y, Ogawa A, Sasakawa S
JournalThromb Res
PubMed ID2499076
'The effects of cytoplasmic acidification by nigericin, monovalent cation ionophore, on platelet activation induced by arachidonic acid (AA) were examined. AA-induced aggregation and TXB2 formation were inhibited with decreasing cytoplasmic pH (pHc). Although TXA2 receptor agonists (STA2, U-46619)-induced aggregation of some platelet samples were suppressed by nigericin, the suppression did ... More
ATP-dependent and DCCD-insensitive Cl- uptake by membrane vesicles from the rat brain plasma membrane fractions.
AuthorsShiroya T, Cragoe EJ, Inagaki C
JournalBiochem Biophys Res Commun
PubMed ID2719675
'ATP-dependent Cl- uptake by membrane vesicles from the rat brain plasma membrane fractions was not affected by the addition of 40 mM of K+, Na+ or HCO3- to the assay medium. Na+ and K+ did not alter the uptake even in the presence of a K+ ionophore, valinomycin (10 microM), ... More
The ionophore nigericin transports Pb2+ with high activity and selectivity: a comparison to monensin and ionomycin.
AuthorsHamidinia SA, Tan B, Erdahl WL, Chapman CJ, Taylor RW, Pfeiffer DR
JournalBiochemistry
PubMed ID15595852
'The K(+) ionophore nigericin is shown to be highly effective as an ionophore for Pb(2+) but not other divalent cations, including Cu(2+), Zn(2+), Cd(2+), Mn(2+), Co(2+), Ca(2+), Ni(2+), and Sr(2+). Among this group a minor activity for Cu(2+) transport is seen, while for the others activity is near or below ... More
Extracellular ATP-induced acidification leads to cytosolic calcium transient rise in single rat cardiac myocytes.
AuthorsPucéat M, Clément O, Scamps F, Vassort G
JournalBiochem J
PubMed ID2001251
'The origin of the increase in cytosolic free Ca2+ concentration ([Ca2+]i) induced by extracellular ATP was investigated in single isolated cardiac myocytes loaded with indo-1. The nucleotide added at a concentration of 10 microM triggers a few Ca2+ spikes, followed by a cluster of Ca2+ oscillations, increasing [Ca2+]i to around ... More
Transport of the cationic fluorochrome rhodamine 123 in an insect's Malpighian tubule: indications of a reabsorptive function of the secondary cell type.
AuthorsMeulemans W, De Loof A
JournalJ Cell Sci
PubMed ID1629249
'The pathway of rhodamine 123 was examined after injection into Sarcophaga flies and after in vitro labeling of the Malpighian tubules. After in vitro labeling the primary cells only retained this potential-sensitive dye for a short period while all secondary cells accumulated the dye from the tubule lumen. In vivo ... More
Cell shrinkage regulates Src kinases and induces tyrosine phosphorylation of cortactin, independent of the osmotic regulation of Na+/H+ exchangers.
AuthorsKapus A, Szászi K, Sun J, Rizoli S, Rotstein OD
JournalJ Biol Chem
PubMed ID10075710
'The signaling pathways by which cell volume regulates ion transporters, e.g. Na+/H+ exchangers (NHEs), and affects cytoskeletal organization are poorly understood. We have previously shown that shrinkage induces tyrosine phosphorylation in CHO cells, predominantly in an 85-kDa band. To identify volume-sensitive kinases and their substrates, we investigated the effect of ... More
Reduced capacitative calcium entry correlates with vesicle accumulation and apoptosis.
'A preneoplastic variant of Syrian hamster embryo cells, sup(+), exhibits decreased endoplasmic reticulum calcium levels and subsequently undergoes apoptosis in low serum conditions (Preston, G. A., Barrett, J. C., Biermann, J. A., and Murphy, E. (1997) Cancer Res. 57, 537-542). This decrease in endoplasmic reticulum calcium appears to be due, ... More
Parathyroid hormone depresses cytosolic pH and DNA synthesis in osteoblast-like cells.
AuthorsReid IR, Civitelli R, Avioli LV, Hruska KA
JournalAm J Physiol
PubMed ID2839040
'It has recently become apparent that a number of hormones and growth factors modulate cytosolic pH (pHi), and there is some evidence that this in turn may influence cell growth. We have examined the effects of parathyroid hormone (PTH) on both these parameters in an osteoblast-like cell line, UMR 106. ... More
Simultaneous flow cytometric measurements of thrombin-induced cytosolic pH and Ca2+ fluxes in human platelets.
AuthorsDavies TA, Weil GJ, Simons ER
JournalJ Biol Chem
PubMed ID2365684
'Human platelets exhibit an extremely rapid increase in cytoplasmic Ca2+ concentrations ((Ca2+]in) and a dose-dependent cytoplasmic pH change ((pH]in) upon thrombin stimulation. A cytoplasmic alkalinization, maximal by 60 s, is preceded by a very rapid acidification, which is masked by the alkalinization when saturating thrombin doses are used. Using the ... More
14-3-3 Binding to Na+/H+ exchanger isoform-1 is associated with serum-dependent activation of Na+/H+ exchange.
AuthorsLehoux S, Abe Ji Florian JA, Berk BC
JournalJ Biol Chem
PubMed ID11279064
'Na(+)/H(+) exchanger isoform-1 (NHE1), the ubiquitous form of the Na(+)/H(+) exchanger, has increased activity in hypertensive patients and in animal models of hypertension. Furthermore, NHE1 is activated in cells stimulated with growth factors. We showed previously that activation of the exchanger is dependent on phosphorylation of serine 703 (Ser(P)(703)) by ... More
Leukotriene D4-induced increases in the cytoplasmic pH of human myelocytic leukocytes.
AuthorsBaud L, Healy A, Cragoe EJ, Goetzl EJ, Koo CH
JournalJ Cell Physiol
PubMed ID2842349
'The effects of leukotriene D4 on the intracellular pH of human myelocytes, derived from cultured HL-60 cells by dimethylsulfoxide-induced differentiation, were quantified with the fluorescent indicator 2'',7''-bis-(2-carboxy-ethyl)-5,6-carboxyfluorescein. Leukotriene D4, but not C4 or E4, increased intracellular pH optimally by 3 min with a half-maximal effect at 1-2 nM. The increases ... More
Role of Na+/H+ exchange by interferon-gamma in enhanced expression of JE and I-A beta genes.
AuthorsPrpic V, Yu SF, Figueiredo F, Hollenbach PW, Gawdi G, Herman B, Uhing RJ, Adams DO
JournalScience
PubMed ID2541500
'The rapid transductional sequences initiated by interferon-gamma (IFN-gamma) on binding to its receptor regulate functional and genomic responses in many cells but are not well defined. Induction of macrophage activation is an example of such functional and genomic changes in response to IFN-gamma. Addition of IFN-gamma to murine macrophages, at ... More
A developmentally regulated Na-H exchanger in Dictyostelium discoideum is necessary for cell polarity during chemotaxis.
AuthorsPatel H, Barber DL
JournalJ Cell Biol
PubMed ID15851518
'Increased intracellular H(+) efflux is speculated to be an evolutionarily conserved mechanism necessary for rapid assembly of cytoskeletal filaments and for morphological polarity during cell motility. In Dictyostelium discoideum, increased intracellular pH through undefined transport mechanisms plays a key role in directed cell movement. We report that a developmentally regulated ... More
A delayed and sustained rise of cytosolic calcium is elicited by oxidized LDL in cultured bovine aortic endothelial cells.
AuthorsNègre-Salvayre A, Fitoussi G, Réaud V, Pieraggi MT, Thiers JC, Salvayre R
JournalFEBS Lett
PubMed ID1544476
'Bovine aortic endothelial cells (BAEC) pulsed for 5 h with mildly oxidized low density lipoproteins (LDL), exhibited a broad, sustained and high peak of [Ca2+]i occurring several hours after the end of the pulse and reaching very high [Ca2+]i values (around 2500-3000 nmol/l) and a concomitant drop of cytosolic pH ... More
Use of rhodamine 123 to investigate alterations in mitochondrial activity in isolated mouse liver mitochondria.
AuthorsO'Connor JE, Vargas JL, Kimler BF, Hernandez-Yago J, Grisolia S
JournalBiochem Biophys Res Commun
PubMed ID3348796
'The fluorescent dye Rhodamine-123, which selectively stains mitochondria depending on the mitochondrial membrane potential, was used with flow cytometry to evaluate alterations in activity of mitochondria isolated from mouse liver. Under in vitro conditions, with succinate and ADP present in the buffer, mitochondrial activity was affected by a variety of ... More
Effects of angiotensin II and vasopressin on intracellular pH of glomerular mesangial cells.
AuthorsGanz MB, Boyarsky G, Boron WF, Sterzel RB
JournalAm J Physiol
PubMed ID3381882
'We investigated changes in intracellular pH (pHi) of cultured rat glomerular mesangial cells (MCs) exposed to angiotensin II (ANG II) and arginine vasopressin (AVP). pHi of quiescent MCs, passage 2-5, and grown on glass cover slips, was assessed by spectrofluorometry using the pH-sensitive dye, 2,7-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF). The steady-state pHi of ... More
pH dependence and compartmentalization of zinc transported across plasma membrane of rat cortical neurons.
AuthorsColvin RA
JournalAm J Physiol Cell Physiol
PubMed ID11788343
'In this study, Zn(2+) transport in rat cortical neurons was characterized by successfully combining radioactive tracer experiments with spectrofluorometry and fluorescence microscopy. Cortical neurons showed a time-dependent and saturable transport of (65)Zn(2+) with an apparent affinity of 15-20 microM. (65)Zn(2+) transport was pH dependent and was decreased by extracellular acidification ... More
Sodium-proton exchange in human ileal brush-border membrane vesicles.
AuthorsRamaswamy K, Harig JM, Kleinman JG, Harris MS, Barry JA
JournalBiochim Biophys Acta
PubMed ID2543457
'This study examines the characteristics of Na+ and H+ transport as well as Na+-H+ exchange in human ileal brush-border membrane vesicles from organ donor intestine. 22Na+ uptake into vesicles and the fluorescence quenching of Acridine orange were employed to measure Na+ and H+ transport, respectively. Concentrative uptake of 22Na+ (4-fold ... More
APB suppresses synaptic input to retinal horizontal cells in fish: a direct action on horizontal cells modulated by intracellular pH.
AuthorsTakahashi K, Copenhagen DR
JournalJ Neurophysiol
PubMed ID1352805
'1. Membrane potentials and cone-driven light responses were recorded from the H1-type horizontal cells in isolated retinas. Membrane potentials and intracellular pH were recorded also in enzymatically dissociated solitary horizontal cells. 2. In isolated retinas the glutamate analogue 2-amino-4-phosphonobutyrate (APB) hyperpolarized horizontal cells and reduced their light responses in a ... More
Functional characterization of a cloned human kidney Na+:HCO3- cotransporter.
AuthorsAmlal H, Wang Z, Burnham C, Soleimani M
JournalJ Biol Chem
PubMed ID9642239
'Functional properties of a cloned human kidney Na+:HCO3- cotransporter (NBC-1) were studied in cultured HEK-293 cells that were transiently transfected with NBC-1 cDNA. The Na+:HCO3- cotransporter activity was assayed as the Na+ and HCO3-dependent pHi recovery from intracellular acidosis with the use of the pH-sensitive dye 2'',7''-bis(2-carboxyethyl)-5(6)-carboxyfluorescein. In acid-loaded cells ... More
Antibiotic-mediated transport of alkali ions across lipid barriers.
AuthorsPressman BC, Harris EJ, Jagger WS, Johnson JH
JournalProc Natl Acad Sci U S A
PubMed ID4230180
Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ.
AuthorsThomas JA, Buchsbaum RN, Zimniak A, Racker E
JournalBiochemistry
PubMed ID36128
Fluorescence dye as monitor of internal pH in Escherichia coli cells.
AuthorsShechter E, Letellier L, Simons ER
JournalFEBS Lett
PubMed ID7042384
Control of coated-pit function by cytoplasmic pH.
AuthorsSandvig K, Olsnes S, Petersen OW, Van Deurs B
JournalMethods Cell Biol
PubMed ID2575206
Intracellular ion activities and membrane transport in parietal cells measured with fluorescent dyes.
AuthorsNegulescu PA, Machen TE
JournalMethods Enzymol
PubMed ID2074799
The purpose of this chapter is to summarize the methods we have usedfor measuring intracellular pH with BCECF, intracellular calcium with fura-2 and intracellular sodium with SBFI in single, identified cells of gastricglands isolated from rabbits. ... More
Activation of Na+/H+ exchange in human platelets stimulated by thrombin and a phorbol ester.
AuthorsSiffert W, Siffert G, Scheid P
JournalBiochem J
PubMed ID3566711
We have investigated changes in cytoplasmic pH (pHi) in activated human platelets, using the fluorescent probe 2,7-biscarboxyethyl-5(6)-carboxyfluorescein. Stimulation of platelets by thrombin or 12-O-tetradecanoylphorbol 13-acetate increased pHi by about 0.11 pH unit above the resting value. This increase in pHi depended on the presence of external Na+ and was inhibited ... More
Cytoplasmic pH in the action of epidermal growth factor (EGF) in cultured porcine thyroid cells.
AuthorsTakasu N, Nagasawa Y, Komiya I, Yamada T, Shimizu Y
JournalBiochem Biophys Res Commun
PubMed ID2848521
We demonstrate measurement of cytoplasmic pH (pHi), using 2',7'-bis(2-carboxyethyl)-5 (and 6-) carboxyfluorescein (BCECF), and internalized fluorescent pHi indicator, in thyroid cells. Using cultured porcine thyroid cells, we studied the effects of epidermal growth factor (EGF) on pHi and [3H] thymidine incorporation; 10 nM EGF alkalinizes thyroid cells and stimulates thymidine ... More
Evaluation of changes in cytoplasmic pH in thrombin-stimulated human platelets.
AuthorsDavies TA, Dunn JM, Simons ER
JournalAnal Biochem
PubMed ID3434788
Thrombin causes a dose-dependent cytoplasmic alkalinization of normal human platelets. The pH probe 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) permits an easier and more accurate quantitation of the kinetics of this change previously measured with 9-aminoacridine and 6-carboxyfluorescein (6-CF). We report here a modification of the previously published 6-CF technique and confirm the thrombin-induced ... More
An increase in intracellular pH is a general response of promonocytic cells to differentiating agents.
AuthorsLadoux A, Damais C, Krawice I, Abita JP, Frelin C
JournalFEBS Lett
PubMed ID2455658
Intracellular pH (pHi) was measured in HL60 and U937 cells before and after differentiation into monocyte-macrophage like cells. 12-O-Tetradecanoyl phorbol-13-acetate (PMA), butyrate, interferon, retinoic acid and 1,25-dihydroxyvitamin D3 all increased pHi. The increases elicited were rapid with PMA, much slower with retinoic acid and interferon and still slower with 1,25-dihydroxyvitamin ... More
Cytosolic calcium, oxygen consumption and the intracellular pH of stimulated neutrophils.
AuthorsNasmith PE, Grinstein S
JournalBiosci Rep
PubMed ID2840131
The cytoplasmic pH undergoes a biphasic change when neutrophils are activated. The role of Ca2+ in initiating these changes was investigated. No correlation was found between the increased cytosolic [Ca2+] and the stimulation of the Na+/H+ antiport. Similarly, the cytoplasmic acidification elicited by activation in Na+-free media was found to ... More
Intracellular pH of halobacteria can be determined by the fluorescent dye 2', 7'-bis(carboxyethyl)-5(6)-carboxyfluorescein.
AuthorsTsujimoto K, Semadeni M, Huflejt M, Packer L
JournalBiochem Biophys Res Commun
PubMed ID3415676
Determination of the internal pH of halobacterial cells grown in 4M salt solution has proven to be a difficult problem. We now report the steady state cytosolic pH of Halobacterium halobium S-9 to be 7.2. Intracellular pH was determined after the cells were loaded with the membrane permeable precursor of ... More