The effects of ionophores on the fluorescence of the cation 3,3'-dipropyloxadicarbocyanine in the presence of pigeon erythrocytes, erythrocyte 'ghosts' or liposomes

• Published in: Biochemical journal Vol. 168; no. 1; pp. 81 - 90
• Main Authors: Kimmich, G A, Philo, R D, Eddy, A A
• Format: Journal Article
• Language: English
• Published: England 15-10-1977
• Subjects: Animals; Carbocyanines; Columbidae; Erythrocyte Membrane - drug effects; Erythrocyte Membrane - metabolism; Erythrocytes - drug effects; Erythrocytes - metabolism; Fluorescence; Fluorescent Dyes; Gramicidin - pharmacology; Ionophores - pharmacology; Liposomes - metabolism; Membrane Potentials - drug effects; Models, Biological; Nigericin - pharmacology; Quinolines; Valinomycin - pharmacology
• ISSN: 0264-6021; 1470-8728
• DOI: 10.1042/bj1680081

1. Pigeon erythrocytes, resealed lysed erythrocytes or liposomes derived from erythrocyte lipids were suspended in solutions containing up to 2 micrometer-3,3'-dipropyloxadicarbocyanine iodide. Gramicidin, valinomycin, nigericin or carbonyl cyanide p-trifluoromethoxy-phenylhydrazone, or combinations of these, were used to induce electrical diffusion potentials dependent on Na+, K+ or protons. In each instance hyperpolarization of the cell membrane lowered the fluorescence of the cell suspension, a process that was completed in about 1 min. Subsequent depolarization caused an increase in fluorescence. 2. Quenching of the fluorescence of the cell suspension appeared to be due to the reversible binding of the dye to the cells. Much larger amounts of dye were bound, both to the intact and to the resealed erythrocytes, than would be expected if partitioning of the dye cation followed the Nernst equation. The dependence of the binding on the extracellular dye concentration was studied in the presence and absence of valinomycin. The results were consistent with the suggestion of Sims, Waggoner, Wang & Hoffman [(1974) Biochemistry 13, 3315-3330] that the dye was bound at both membrane surfaces and that, at low dye concentrations, hyperpolarizing the cells promoted dye binding at the inner membrane surface. 3. The applications of the technique are limited by the circumstance that the direct effect of the electric field on the uptake of the dye into the cells is amplified by a binding process that may be affected by other physiological variables.