Video-imaging micro-fluorometric assessment of luminal chloride/bicarbonate exchange activity in Madin-Darby canine kidney cells: influence of cell density, 4,4'-diisothiocyano-2,2'-disulfonic stilbene and acetazolamide.
AuthorsEbner S, Marin-Grez M
JournalBiol Chem Hoppe Seyler
PubMed ID8828818
'To investigate whether or not MDCK cells may be used as a model for beta-intercalated cells, we studied: (1) The effect of luminal [Cl-]0 changes on pHi measured by video-imaging micro-fluorometry, (2) the influence of the inhibitor 4,4''-diisothiocyano-2,2''-disulfonic stilbene (DIDS) on anion-exchange activity, and (3) the effect of acetazolamide on ... More
Continuous recording of intramitochondrial pH with fluorescent pH indicators: novel probes and limitations of the method.
AuthorsZólkiewska A, Czyz A, Duszynski J, Wojtczak L
JournalActa Biochim Pol
PubMed ID8212962
'In addition to 2'',7''-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) used so far to monitor intramitochondrial pH, two other fluorescent pH indicators, 4'',5''-dimethyl-5(6)-carboxyfluorescein (DMCF) and carboxyseminaphthofluorescein (carboxy-SNAFL-1), were applied for this purpose. These probes are taken up by isolated rat liver mitochondria in form of diacetate esters, hydrolyzed within mitochondria to free acids, and respond ... More
Multiparameter flow cytometric analysis of a pH sensitive formyl peptide with application to receptor structure and processing kinetics.
AuthorsFay SP, Habbersett R, Domalewski MD, Posner RG, Houghton TG, Pierson E, Muthukumaraswamy N, Whitaker J, Haugland RP, Freer RJ
JournalCytometry
PubMed ID8168401
'Environmentally sensitive molecules have many potential cellular applications. We have investigated the utility of a pH sensitive ligand for the formyl peptide receptor, CHO-Met-Leu-Phe-Phe-Lys (SNAFL)-OH (SNAFL-seminaphtho-fluorescein), because in previous studies (Fay et al.: Biochemistry 30:5066-5075, 1991) protonation has been used to explain the quenching when the fluoresceinated formyl pentapeptide ligand ... More
Quantitative pH imaging in cells using confocal fluorescence lifetime imaging microscopy.
AuthorsSanders R, Draaijer A, Gerritsen HC, Houpt PM, Levine YK
JournalAnal Biochem
PubMed ID7573951
'The pH-sensitive probe carboxy SNAFL-1 can be used for imaging using ratiometric and fluorescence lifetime techniques. The former method suffers from the drawback that quantitative pH imaging in cells requires a time-consuming and cumbersome calibration procedure. In contrast, straightforward calibrations in buffer suffice for fluorescence lifetime imaging. This is illustrated ... More
Video-imaging microfluorometry identifies alpha- and beta-like cell types in Madin-Darby canine kidney monolayers.
AuthorsEbner S, Marin-Grez M
JournalKidney Int Suppl
PubMed ID9736270
'On the basis of intracellular, accumulation of c-SNAFL-2, we have identified three cell subtypes in Madin-Darby canine kidney (MDCK) monolayers. Highly fluorescent cells (HFC) have a high intracellular pH (pHi, whereas cells with medium fluorescence (MFC) have low pHi when perfused with buffer containing 125 mM Cl-. HFC express a ... 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
Fluorescence lifetime spectroscopy for pH sensing in scattering media.
AuthorsKuwana E, Sevick-Muraca EM
JournalAnal Chem
PubMed ID14632153
'Fluorescence lifetime spectroscopy in the presence of tissuelike scattering is demonstrated from measurements of phase and modulation ratio as a function of modulation frequency using a pH-sensitive dye, Carboxy Seminaphthofluorescein-1 (C-SNAFL-1). From the optical diffusion equation describing the propagation and generation of fluorescence within solutions of 0.5 microM C-SNAFL-1 containing ... More
Regulation of cytosol-nucleus pH gradients by K+/H+ exchange mechanism in the nuclear envelope of neonatal rat astrocytes.
AuthorsMasuda A, Oyamada M, Nagaoka T, Tateishi N, Takamatsu T
JournalBrain Res
PubMed ID9756998
In order to study the subcellular heterogeneity of intracellular H+ concentration in reactive astrocytes, the pH in the nucleus and cytosol of cultured astrocytes was measured using a confocal laser scanning microscope (CLSM) and pH indicator dye, 5'(and 6')-carboxyseminaphthofluorescein (carboxy SNAFL-1). The change in intracellular pH was indexed by the ... More
Intracellular alkalinization induced by bradykinin sustains activation of the constitutive nitric oxide synthase in endothelial cells.
AuthorsFleming I, Hecker M, Busse R
JournalCirc Res
PubMed ID7514511
The transient increase in [Ca2+]i in endothelial cells after stimulation with bradykinin can account for the initiation but not the sustained production of nitric oxide (NO). Therefore, we investigated whether this sustained activation of the constitutive NO synthase (cNOS) could be mediated by an increase in pHi, which is induced ... More
Extracellular and intracellular alkalinization and the constriction of rat cerebral arterioles.
AuthorsApkon M, Boron WF
JournalJ Physiol
PubMed ID7623289
1. Direct observations of perfused cerebral arterioles in vivo and in vitro have demonstrated that alkalinization of blood or cerebrospinal fluid (CSF) causes arteriolar constriction. Inasmuch as such alkalinizations lead to increases in intracellular pH (pHi) as well as interstitial pH (pHo), it is possible that increases in either pHi ... More