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L Reuss

Publications and source records attributed to L Reuss.

At least 109 records · Page 6Linked to original sources

Dual effects of amphotericin B on ion permeation in toad urinary bladder epithelium.

The mechanisms of action of amphotericin B on the electrical properties of the toad urinary bladder epithelium were studied with microelectrode techniques. Cell membrane and transepithelial electrical potentials and resistances were measured in the absence and in the presence of the drug during exposure to bathing mediums of different ionic compositions. As observed previously by other investigators, amphotericin B produces a dramatic decrease of transepithelial electrical resistance (Rt) and an increase of the rate of sodium transport. Our results indicate that the effect of the drug on Rt depends in part on an increase in Na conductance across the luminal cell membrane (amiloride-insensitive), but is caused mainly by an increase of ionic conductances (with the sequence GK greater than GNa greater than G choline greater than GCI) at a site in parallel with the impaled cells (i.e., across a cell type not investigated by the microelectrode measurements or across the paracellular pathway.

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Mechanisms of voltage transients during current clamp in Necturus gallbladder.

Microelectrode techniques were employed to study the mechanisms of the transepithelial voltage transients (deltaVms) observed during transmural current clamps in the isolated Necturus gallbladder. The results indicate that: a) part of deltaVms is due to a transepithelial resistance change (deltaRt), and part to a tissue emf change. b) deltaRt is entirely caused by changes of the resistance of the paracellular pathway. At all current densities employed, the measured changes are probably due to changes in both fluid conductivity and width of the lateral intercellular spaces. At high currents, in addition to the effects on the lateral spaces, the resistance of other elements of the pathway (probably the limiting junction) drops, regardless of the direction of the current. c) The magnitude and polarity of the deltaRt-independent transepithelial and cell membrane potential transients indicate that the largest emf change takes place at the basolateral membrane (deltaEb), with smaller changes at the luminal membrane (deltaEa) and the paracellular (shunt) pathway (deltaEs). It is shown that two-thirds of the transient are caused by deltaEs, and one-third by delta(Eb--Ea). deltaEs can be explained by a diffusion potential generated by a current-dependent NaCl concentration gradient across the tissue. deltaEa and deltaEb are caused by [K] changes, mainly at the unstirred layer in contact with the basolateral membrane.

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Effects of luminal hyperosmolality on electrical pathways of Necturas gallbladder.

The electrical properties of the transepithelial pathways of Necturaus gallbladder were studied with intracellular microelectrode techniques under control conditions and after exposure to hyperosmotic mucosal bathing media (addition of sucrose or other solutes). Doubling mucosal osmolality produces a large increase in the resistance of the intercellular shunt pathway (from 420 +/- 50 to 700 +/- 70 omega - cm2, P is less than 0.001) and a significant decrease in the resistance of the apical membrane of the cells (from 3,510 +/- 420 to 1,540 +/- 380 omega p cm2, P is less than 0.001). The basolateral membrane resistance remains unchanged. Both the apical and basolateral membranes depolarize (from 59.5 +/- 1.8 to 36.6 +/- 2.8 mV, P less than 0.0001, and from 61.8 +/- 1.7 to 32.2 +/- 3.0 mV,P is less than 0.001, respectively). The transepithelial diffusion potential resulting from NaCl concentration gradients due to the osmotic water flow does not explain the cell potential changes. Mucosal solution ion substitution experiments demonstrate increases of gk, gcl, and gna (ca. 2.5-fold, 4-fold, and 7-fold, respectively). Therefore, cell depolarization appears to be produced mainly by a reduction of apical membrane K permselectivity. The increase in the resistance of the shunt is attributable to reduction of the width of the lateral intercellular spaces, because of the osmotic serosa-to-mucosa water flow.

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Electrical properties of the cellular transepithelial pathway in Necturus gallbladder. I. Circuit analysis and steady-state effects of mucosal solution ionic substitutions.

Microelectrode techniques were employed to measure the electrical resistance of the cell membranes and the shunt pathway, and the equivalent electromotive forces (EMF's) at both cell borders in Necturus gallbladder epithelium. The cell is, on the average, 57 mV negative to the mucosal solution and 59 mV negative to the serosal solution. The transepithelial potential (Vms) ranges from 0.5 to 5 mV, serosal solution positive. Assuming that the shunt EMF (Vs) is zero with standard Ringer's bathing oth sides of the tissue, both cell membrane EMF's are oriented with the negative pole toward the cell interior and are 39.9 +/- 3.6 mV (apical, Va), and 69.4 +/- 1.8 mV (basal-lateral, Vb)...

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Electrical properties of the cellular transepithelial pathway in Necturus gallbladder. II. Ionic permeability of the apical cell membrane.

Microelectrode techniques were employed to study the ionic permeability of the apical cell membrane of Necturus gallbladder epithelium. Results obtained from continuous records in single cells, and from several cellular impalements shortly after a change in solution, were similar and indicate that both the apical membrane equivalent electromotive force (Va) and electrical resistance (Ra) strongly depend on external [K]. Cl substitutions produced smaller effects, while the effects of Na substitutions with N-methyl-D-glucamine on both Va and Ra were minimal. These results indicate that the permeability sequence of the apical membrane is PKgreater thanPClgreater than PNa. From the calculated absolute value of PNa it is possible to estimate the diffusional Na flux from the mucosal solution into the cells (from the cell potential and an assumed intracellular Na concentration). The calculated flux is roughly three orders of magnitude smaller than the measured net transepithelial flux in this tissue and in gallbladders of other species. Thus, only a minimal portion of Na entry can be attributed to independent diffusion. From estimations of the electrochemical potential gradient across the apical membrane, Cl transport at that site must be active. At the serosal cell membrane, Na transport takes place against both chemical and electrical potentials, while a significant portion of the Cl flux can be passive, if this membrane has a significant Cl conductance. The changes in shunt electromotive force and in transepithelial potential after mucosal substitutions were very similar, indicating that transepithelial bi-ionic potentials yield appropriate results on the properties of shunt pathway.

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Effects of changes in the composition of the mucosal solution on the electrical properties of the toad urinary bladder epithelium.

By the use of microelectrode techniques, the potential profile and the electrical resistances of the cellular and shunt pathways across the toad urinary bladder epithelium were measured under control conditions and after exposing the mucosal side to solutions of low and high NaCl concentrations and osmolalities. The resistance of the shunt pathway increases at low NaCl concentration (even if the osmolality is kept constant), and decreases at high NaCl concentration (by a nonspecific osmotic mechanism). The inverse relationship between mucosal NaCl concentration and shunt resistance suggests a regulatory mechanism of net sodium transport by reduction of the passive blood-to-urine sodium flux at low urinary sodium concentrations. In addition, the transepithelial potential and the potentials at both cell borders fall in both low and high mucosal NaCl, and the magnitude of these changes is such that they cannot be explained by changes in the shunt pathway alone.

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Effects of changes in the composition of the serosal solution on the electrical properties of the toad urinary bladder epithelium.

1. The potential profile and the cellular and paracellular transepithelial resistances of the toad urinary bladder were measured, by means of micro-electrode techniques, as functions of the osmolality of the serosal solution. 2. Reductions in serosal osmolality (that increase the rate of active sodium transport) produced proportional decreases in the electrical resistances of the apical and basal-lateral cell membranes, while the changes in resistance of the paracellular pathway were more complex. The apical membrane potential increased. 3. Increases in serosal osmolality (that decrease sodium transport) produced increases in the electrical resistances of both cell membranes, and moderate reduction in the paracellular resistance. The polarity of the apical membrane potential reversed. 4. These results indicate that reductions in serosal solution osmolality stimulate sodium transport by increasing both the sodium permeability of the luminal cell membrane (thus increasing sodium entry), and the electromotive force generated at the serosal border of the cell, thus enhancing the rate of sodium pumping. Conversely, increases in osmolality reduced sodium transport by reducing both the sodium permeability of the luminal membrane and the serosal membrane electromotive force.

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Passive electrical properties of toad urinary bladder epithelium. Intercellular electrical coupling and transepithelial cellular and shunt conductances.

The electrical resistances of the transcellular and paracellular pathways across the toad urinary bladder epithelium (a typical "tight" sodium-transporting epithelium) were determined by two independent sets of electrophysiological measurements: (a) the measurement of the total transepithelial resistance, the ratio of resistance of the apical to the basal cell membrane, and cable analysis of the voltage spread into the epithelium; (b) the measurement of the total transepithelial resistance and the ratio of resistances of both cell membranes before and after replacing all mucosal sodium with potassium (thus, increasing selectively the resistance of the apical membrane). The results obtained with both methods indicate the presence of a finite transepithelial shunt pathway, whose resistance is about 1.8 times the resistance of the transcellular pathway. Appropriate calculations show that the resistance of the shunt pathway is almost exclusively determined by the zonula occludens section of the limiting junctions. The mean resistance of the apical cell membrane is 1.7 times that of the basal cell membrane. The use of nonconducting materials on the mucosal side allowed us to demonstrate that apparently all epithelial cells are electrically coupled, with a mean space constant of 460 microm, and a voltage spread consistent with a thin sheet model.

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