IgE receptor-mediated depolarization of rat basophilic leukemia cells measured with the fluorescent probe bis-oxonol.
Receptor-mediated changes in plasma membrane potential were recorded in rat basophilic leukemia (RBL) cells with the potential-sensitive fluorescent indicator bis-oxonol. Depolarization of the mitochondria with metabolic inhibitors was not detected by bis-oxonol, suggesting that only potential changes across the plasma membrane were being measured. The resting membrane potential of RBL cells was largely generated by the equilibrium distribution of K+ and not through electrogenic activity of the sodium pump. Depolarization was maintained as long as IgE receptors remained aggregated. We believe that at physiologic calcium concentrations a large portion of the measured potential change may be due to calcium influx across the plasma membrane. Prevention of calcium influx by lanthanum, disruption of aggregated receptors, or prior depolarization in a high K+ saline solution completely inhibited the antigen-induced depolarization. The time course of the antigen-stimulated increase in bis-oxonol fluorescence was similar, but not identical, to the antigen-stimulated rise in cytoplasmic free ionized calcium measured with fura-2. Antigen-stimulated depolarization was inhibited by removing both calcium and sodium and could be restored by the addition of either ion. Reduction of total cellular adenosine triphosphate inhibited depolarization in response to antigen stimulation.
D000255:Adenosine Triphosphate / Q000502:physiology; D000818:Animals; D001491:Basophils / Q000502:physiology*; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D007073:Immunoglobulin E / Q000502:physiology*; D007152:Immunologic Capping; D007811:Lanthanum / Q000494:pharmacology; D007942:Leukemia, Experimental / Q000503:physiopathology; D008560:Membrane Fluidity; D008564:Membrane Potentials; D011188:Potassium / Q000502:physiology; D051381:Rats; D011961:Receptors, Fc / Q000502:physiology*; D017455:Receptors, IgE; D011971:Receptors, Immunologic / Q000502:physiology*; D013050:Spectrometry, Fluorescence; D013858:Thiobarbiturates
D011961:Receptors, Fc; D017455:Receptors, IgE; D011971:Receptors, Immunologic; D013858:Thiobarbiturates; D007073:Immunoglobulin E; C025284:bis(1,3-diethylthiobarbiturate)trimethineoxonol; D007811:Lanthanum; D000255:Adenosine Triphosphate; D011188:Potassium