nystatin-a1 and gluconic-acid

Dihydropyridine-sensitive calcium currents (I(Ca)) in photoreceptors are unusual in that they can be inhibited by reductions in extracellular chloride. The present study examined whether I(Ca) in retinal bipolar cells, which as in photoreceptors mediates sustained neurotransmission, is also inhibited by reductions in chloride. Nystatin-perforated patch, whole cell recordings were obtained from bipolar cells in a retinal slice preparation of larval tiger salamander. In the presence of Ba(2+), voltage steps above -40 mV evoked sustained inward currents, which were enhanced by the dihydropyridine, (-)BayK8644, and inhibited by nisoldipine. Similar to photoreceptors, replacing Cl(-) with gluconate or CH(3)SO(4) inhibited bipolar cell I(Ca) and produced a negative shift in the current/voltage relationship. Thus, sensitivity to Cl(-) may be a more general property of L-type Ca(2+) channel subtypes that mediate sustained neurotransmission.

Topics: 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; Ambystoma; Animals; Barium; Calcium Channels, L-Type; Chlorides; Dihydropyridines; Gluconates; In Vitro Techniques; Larva; Membrane Potentials; Nisoldipine; Nystatin; Patch-Clamp Techniques; Photoreceptor Cells, Vertebrate; Retina

In the isolated bullfrog cornea epithelium, under short-circuit conditions the regulation of the K permeability of the basolateral membrane was studied with conventional and K-selective microelectrodes in Cl-free Ringers. In Cl-free Ringers, the transcellular current is less than 1 microA/cm2, allowing estimation of the basolateral membrane electromotive force from measurements of the membrane voltage (Vsc). The apparent basolateral membrane K conductance was determined from measurements of the effects of single ion substitutions of K for Na on the Vsc. An increase of K from 2.5 to 25 mM on the stromal side depolarized the membrane voltage by 29 mV, whereas additional increases to 56 and 100 mM resulted in depolarizations consistent with a Nernstian prediction. In the range between 25 and 56 mM K, these decreases in membrane voltage were smaller after either decreasing the stromal-side pH from 8.1 to 7.2 or substitution of sulfate with gluconate. In contrast, preincubation with 0.1 mM ouabain did not change the membrane voltage depolarizations over any of the K ranges between 2.5 and 100 mM. Equivalent circuit analysis, based on the effects of nystatin on the electrical parameters, was used to validate the changes in the apparent basolateral membrane K conductance following increases in [K], substitution of SO4 with gluconate and Na:K pump inhibition. An increase in the [K] to 120 mM decreased the basolateral membrane resistance nearly three-fold, whereas gluconate substitution resulted in a 2.5-fold increase of the basolateral membrane resistance. This resistance increased an additional 2-fold after exposure to 5 mM Ba.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Cell Membrane; Cell Membrane Permeability; Cornea; Epithelium; Gluconates; Membrane Potentials; Nystatin; Ouabain; Potassium; Rana catesbeiana; Sodium-Potassium-Exchanging ATPase

Experiments were performed with intact human red blood cells to determine whether the inhibitory effects of high Cl- concentrations on Cl- exchange are primarily due to interaction at the cytoplasmic or the external surface of the membrane. When internal Cl- was varied from 150 mM to 600 mM Cl- (using the nystatin technique), keeping external Cl- constant at 150 mM (with sucrose added to maintain osmotic balance), Cl- exchange was inhibited almost exactly as much as when both internal and external Cl- were increased from 150 mM to 600 mM. On the other hand, if internal Cl- was maintained constant at 600 mM, variation of external Cl- (with either sucrose, gluconate, or citrate-sucrose mixtures replacing Cl-) had no consistent effect on Cl- exchange. Even if internal Cl- was kept at 150 mM by substitution of gluconate for Cl-, an increase in external Cl- from 150 mM to 600 mM did not significantly inhibit Cl- exchange. Thus the self-inhibitory effects of Cl- seem to be caused primarily by binding to a site at the cytoplasmic side of the membrane. External Br-, on the other hand, did cause a significant inhibition of Cl- exchange. In contrast to the inhibitory effects of Cl- at neutral pH, at very high pH (around pH 11) there is an activation of Cl- exchange at very high Cl- concentrations. This effect, however, depends on binding of Cl- to an external site. Thus there seem to be at least two different low-affinity Cl- binding sites, one at the cytoplasmic side, which inhibits Cl- exchange, and one at the external side, which activates Cl- exchange at high external pH.

Topics: Anion Exchange Protein 1, Erythrocyte; Binding Sites; Biological Transport, Active; Bromine; Cell Membrane; Cell Membrane Permeability; Chlorides; Cytoplasm; Erythrocytes; Gluconates; Humans; Hydrogen-Ion Concentration; Ion Channels; Ion Exchange; Nystatin