Anion transport heterogeneity detected by flow cytometric measurement of NBD-taurine efflux kinetics. 1984

K A Muirhead, and R C Steinfeld, and M C Severski, and P A Knauf

NBD-taurine [N-(7-nitrobenzofuran-4-yl) taurine], a fluorescent substrate for the human erythrocyte anion exchange system, has been used to test the feasibility of making flow cytometric measurements of anion transport in K562 erythroleukemic cells. Cells were preloaded by incubation with 20 microM-2mM NBD-taurine, then diluted 10-30-fold, and efflux was monitored by measuring fluorescence intensity (FL) as a function of time using excitation at 488 nm. The observed rate of decrease in fluorescence was sensitive to temperature and also to phloretin, a compound known to inhibit anion transport and other carrier-mediated transport processes. The coefficient of variation (CV) of the fluorescence distribution increased markedly over the efflux period, suggesting heterogeneity of the K562 population with respect to the rate constant for NBD-taurine efflux. This heterogeneity was also reflected in the upward curvature of a first order plot of log (FLt - FL infinity) versus time. Half-times calculated from initial linear portions of the first-order plots were found to decrease as the loading concentration of NBD-taurine was decreased, as predicted for a saturable transport system. NBD-taurine is not an ideal anion transport substrate for flow cytometric studies. It appears to bind to high-affinity sites within the cells with consequent fluorescence quenching, complicating interpretation of kinetic curves at low concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D010069 Oxadiazoles Compounds containing five-membered heteroaromatic rings containing two carbons, two nitrogens, and one oxygen atom which exist in various regioisomeric forms. Oxadiazole
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D002462 Cell Membrane The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells. Plasma Membrane,Cytoplasmic Membrane,Cell Membranes,Cytoplasmic Membranes,Membrane, Cell,Membrane, Cytoplasmic,Membrane, Plasma,Membranes, Cell,Membranes, Cytoplasmic,Membranes, Plasma,Plasma Membranes
D004915 Leukemia, Erythroblastic, Acute A myeloproliferative disorder characterized by neoplastic proliferation of erythroblastic and myeloblastic elements with atypical erythroblasts and myeloblasts in the peripheral blood. Di Guglielmo's Disease,Erythremic Myelosis,Erythroblastic Leukemia, Acute,Erythroleukemia,Leukemia, Myeloid, Acute, M6,Myeloid Leukemia, Acute, M6,Di Guglielmo Disease,Acute Erythroblastic Leukemia,Acute Erythroblastic Leukemias,Di Guglielmos Disease,Disease, Di Guglielmo,Disease, Di Guglielmo's,Erythremic Myeloses,Erythroblastic Leukemias, Acute,Erythroleukemias,Leukemia, Acute Erythroblastic,Leukemias, Acute Erythroblastic,Myeloses, Erythremic,Myelosis, Erythremic
D005434 Flow Cytometry Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake. Cytofluorometry, Flow,Cytometry, Flow,Flow Microfluorimetry,Fluorescence-Activated Cell Sorting,Microfluorometry, Flow,Cell Sorting, Fluorescence-Activated,Cell Sortings, Fluorescence-Activated,Cytofluorometries, Flow,Cytometries, Flow,Flow Cytofluorometries,Flow Cytofluorometry,Flow Cytometries,Flow Microfluorometries,Flow Microfluorometry,Fluorescence Activated Cell Sorting,Fluorescence-Activated Cell Sortings,Microfluorimetry, Flow,Microfluorometries, Flow,Sorting, Fluorescence-Activated Cell,Sortings, Fluorescence-Activated Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000838 Anions Negatively charged atoms, radicals or groups of atoms which travel to the anode or positive pole during electrolysis. Anion
D013654 Taurine A conditionally essential nutrient, important during mammalian development. It is present in milk but is isolated mostly from ox bile and strongly conjugates bile acids. Taufon,Tauphon,Taurine Hydrochloride,Taurine Zinc Salt (2:1),Taurine, Monopotassium Salt

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